Impact of rumen cannulation surgery on rumen microbiota composition in Hanwoo steers.

Rumen cannulation is a surgical technique used to collect rumen contents from ruminants. However, rumen cannulation surgery may potentially impact the composition of the rumen microbiota. This study aimed to examine the longitudinal alterations in the rumen microbiota composition of Hanwoo steers after cannulation surgery. In this study, eight Hanwoo steers were used; four steers underwent rumen cannulation surgery (cannulation group), while the remaining four were left intact (control group). Rumen samples were collected from all eight steers using the stomach tubing method on the day before surgery (day 0) and on postoperative days 1, 4, 7, 10, 14, 17, 21, 24, and 28, resulting in 80 samples (10 timepoints × 8 animals). The microbiota of all 80 samples were analyzed using 16S rRNA gene amplicon sequencing with Quantitative Insights into Microbial Ecology version 2 (QIIME2). There were no significant differences (p > 0.05) in all major phyla and most major genera representing at least 0.5% of total sequences across all 80 samples between the control and cannulation groups on the preoperative and postoperative days. However, while the alpha diversity indices did not differ (p > 0.05) between the two groups on the preoperative day, they significantly differed (p < 0.05) between the two groups on the postoperative days. Further, the overall microbial distribution based on both unweighted and weighted principal coordinate analysis plots significantly differed (p < 0.05) between the two groups on both the preoperative and postoperative days. Orthogonal polynomial contrasts indicated that major genera and microbial diversity in the cannulation group decreased following surgery but returned to their initial states by postoperative day 28. In conclusion, this study demonstrates that rumen cannulation surgery affects some major taxa and microbial diversity, suggesting that the rumen cannulation method can alter the composition of rumen microbiota in Hanwoo steers.

Modified t-butyl in tetradentate platinum (II) complexes enables exceptional lifetime for blue-phosphorescent organic light-emitting diodes.

In blue phosphorescent dopants, the tetradentate platinum(II) complex is a promising material showing high efficiency and stability in devices. However, metal-metal-to-ligand charge transfer (MMLCT) formation leads to low photo-luminescence quantum yields (PLQYs), wide spectra, and intermolecular interaction. To suppress MMLCT, PtON-tb-TTB and PtON-tb-DTB are designed using theoretical simulation by modifying t-butyl in PtON-TBBI. Both materials effectively suppress MMLCT and exhibit high PLQYs of 99% and 78% in 5 wt% doped film, respectively. The PtON-tb-TTB and PtON-tb-DTB devices have maximum external quantum efficiencies of 26.3% and 20.9%, respectively. Additionally, the PtON-tb-DTB device has an extended lifetime of 169.3 h with an initial luminescence of 1200 nit, which is 8.5 times greater than the PtON-TBBI device. Extended lifetime because of suppressed MMLCT and smaller displacement between the lowest triplet and triplet metal-centered states compared to other dopants. The study provides an effective approach to designing platinum(II) complexes for long device lifetimes.

Gut microbiota alteration with growth performance, histopathological lesions, and immune responses in Salmonella Typhimurium-infected weaned piglets.

Salmonella Typhimurium can cause gastroenteritis in weaned piglets, which are particularly vulnerable to dietary changes and dysfunction of their immature organs. The colonization of S. Typhimurium could disrupt the gut microbiota and increase susceptibility to the bacterium. This study aimed to investigate the alterations of gut microbiota in S. Typhimurium-infected weaned piglets. Ten 49-day-old pigs were divided into two groups: S. Typhimurium-inoculated (ST, n = 6) and negative control (NC, n = 4) groups. The body weight and S. Typhimurium fecal shedding were monitored for 14 days after S. Typhimurium inoculation (dpi). The intestinal tissues were collected at 14 dpi; histopathological lesions and cytokine gene expression were evaluated. The gut microbiome composition and short-chain fatty acid concentrations were analyzed in fecal samples collected at 14 dpi. The average daily gain and gut microbiota alpha diversity in ST group tended to be lower than NC group at 14 dpi. Linear discriminant analysis effect size results showed a significant increase in the abundance of two genera and five species, while a significant decrease was observed in the five genera and nine species within the gut microbiota of ST group. Among the significantly less abundant bacteria in the ST group, Lachnospira eligens and Anaerobium acetethylicum produce acetate and butyrate, and may be considered as key S. Typhimurium infection-preventing bacteria. The overall results provide invaluable information about changes in the gut microbiota of S. Typhimurium-infected weaned piglets, which can be used to develop alternative measures to antibiotics and prevent ST bacterial infection.

Raw potato starch diet supplement in weaned pigs could reduce Salmonella Typhimurium infection by altering microbiome composition and improving immune status.

Backgorund: Salmonella enterica serovar Typhimurium (ST) is one of the causative agents of gastroenteritis in pigs. Pigs fed a diet supplemented with raw potato starch (RPS) have improved gut health by the alteration of the microbiota composition and production of short-chain fatty acids (SCFAs). This study aimed to evaluate the effects of RPS supplementation in reducing infection severity and fecal shedding in ST-infected pigs. Methods: The weaned experimental pigs were divided into two groups: CON (n = 6) fed a corn/soybean-based diet and TRT (n = 6) supplemented with 5% RPS. After 21 d, the pigs were inoculated with ST, and their body weight, clinical signs, and fecal shedding of ST were monitored for 14 d. At 14 d post-inoculation (dpi), the jejunum, cecum, ileum, and colon tissues were collected from euthanized pigs, and histopathological lesions and cytokine gene expression were compared. Additionally, blood samples at 2 dpi were analyzed for gene ontology enrichment. Moreover, the gutmicrobiome was analyzed using 16S rRNA metagenomic sequencing, and the SCFA concentration was measured using gas chromatography. Results: The average daily weight gain was significantly higher in TRT than in CON during the ST infection period; however, histopathological lesion scores were significantly lower in TRT than in CON. The relative abundance of nine genera of butyrate- and acetate-producing bacteria significantly increased in TRT compared with that of only two acetate-producing bacteria in CON. Among the genes involved in the immune response, IL-18 expression level was significantly lower in the jejunum and colon in TRT than in CON. Furthermore, Reg3γ expression was significantly different in the cecum and colon of both groups. Conclusion: The diet supplemented with RPS in weaned pigs could result in predominance of butyrate- and acetate-producing bacteria, reducing the severity of ST infection by improving the immune status.

Pathobiological analysis of african swine fever virus contact-exposed pigs and estimation of the basic reproduction number of the virus in Vietnam.

BACKGROUND: African swine fever (ASF), caused by African swine fever virus (ASFV), is a fatal disease affecting wild and domestic pigs. Since China reported the first ASF outbreak in August 2018, ASFV has swept over the neighbouring Asian countries. However, studies involving experimental pig-to-pig ASFV transmission in Vietnam are lacking. The main objective of this experimental study was to demonstrate the pathobiological characteristics of ASFV contact-exposed pigs and estimate their basic reproduction number (R0) in Vietnam. Fifteen pigs were randomly divided into two groups: experimental (n = 10) and negative control (n = 5) groups. One pig in the experimental group was intramuscularly inoculated with ASFV strain from Vietnam in 2020 and housed with the uninoculated pigs during the study period (28 days). RESULTS: The inoculated pig died 6 days post-inoculation, and the final survival rate was 90.0%. We started observing viremia and excretion of ASFV 10 days post-exposure in contact-exposed pigs. Unlike the surviving and negative control pigs, all necropsied pigs showed severe congestive splenomegaly and moderate-to-severe haemorrhagic lesions in the lymph nodes. The surviving pig presented with mild haemorrhagic lesions in the spleen and kidneys. We used Susceptible-Infectious-Removed models for estimating R0. The R0 values for exponential growth (EG) and maximum likelihood (ML) were calculated to be 2.916 and 4.015, respectively. In addition, the transmission rates (ß) were estimated to be 0.729 (95% confidence interval [CI]: 0.379-1.765) for EG and 1.004 (95% CI: 0.283-2.450) for ML. CONCLUSIONS: This study revealed pathobiological and epidemiological information in about pig-to-pig ASFV transmission. Our findings suggested that culling infected herds within a brief period of time may mitigate the spread of ASF outbreaks.

Multi-organ metastases of primary chondrosarcoma of the mammary gland of a spayed Toy Poodle.

We report the clinical and histopathological findings associated with multi-organ metastases of primary mammary chondrosarcoma in a 12-year-old spayed female Toy Poodle. At post-mortem examination, multifocal, sharply demarcated, grey-white to bright brown, round nodules of variable size were randomly distributed in the lungs, myocardium, liver, pancreas, spleen, intestinal tract and kidneys. Histologically, immature cartilage structures and primitive mesenchymal cells were seen in these organs. Neoplastic cells located in the cartilaginous basophilic extracellular matrix had cytoplasmic vacuolation and round vesicular nuclei and were stained with Safranin O and Alcian blue. To the best of our knowledge, this is the first report of a multi-organ metastatic chondrosarcoma that originated in the mammary gland of a dog.

Johne's disease in two Korean black goats (Capra hircus coreanae) with clinical and subclinical infection: A case report.

Johne's disease (JD) is a chronic enteric infection in ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). JD infection is more difficult to diagnose in goats than cattle because MAP can insidiously affect small ruminants. Few reports have used pathological and molecular diagnosis for cases in Korean black goats. Here, we present our results from two MAP-infected goats. Case 1 was categorized as clinically significant (stage IV), with severe clinical signs and a high antibody titer (S/P ratio, 158.9%). Case 2 was in the subclinical stage (stage II); however, the goat suddenly died without any clinical signs (S/P ratio, 70.9%). DNA from the organ tissues and feces from Case 1 showed a strong positive PCR result for MAP, whereas Case 2 only exhibited a very weak reaction in the fecal sample. Moreover, fecal DNA from both cases was genotyped as C-type MAP using the PCR-REA method. Gastrointestinal organ tissues (jejunum, ileum, colon, and mesenteric lymph nodes) from Case 1 showed moderate-to-severe lesions, and acid-fast bacilli were observed. In contrast, Case 2 showed intact-to-mild pathological lesions, and acid-fast bacilli were detected in the colon, mesenteric lymph nodes, and liver. In addition, Case 2 was co-infected with Corynebacterium pseudotuberculosis, which caused caseous lymphadenitis. This case study provides valuable information regarding the pathological and molecular characteristics of JD-infected Korean black goats. The results highlighted the differences in pathological lesions between clinically and subclinically infected goats, which help veterinarians to develop better strategies to control MAP in goat farms.

Effect of feeding raw potato starch on the composition dynamics of the piglet intestinal microbiome.

OBJECTIVE: Raw potato starch (RPS) is resistant to digestion, escapes absorption, and is metabolized by intestinal microflora in the large intestine and acts as their energy source. In this study, we compared the effect of different concentrations of RPS on the intestinal bacterial community of weaned piglets. METHODS: Male weaned piglets (25-days-old, 7.03±0.49 kg) were either fed a corn/soybean-based control diet (CON, n = 6) or two treatment diets supplemented with 5% RPS (RPS5, n = 4) or 10% RPS (RPS10, n = 4) for 20 days and their fecal samples were collected. The day 0 and 20 samples were analyzed using a 16S rRNA gene sequencing technology, followed by total genomic DNA extraction, library construction, and high-throughput sequencing. After statistical analysis, five phyla and 45 genera accounting for over 0.5% of the reads in any of the three groups were further analyzed. Furthermore, short-chain fatty acids (SCFAs) in the day 20 fecal samples were analyzed using gas chromatography. RESULTS: Significant changes were not observed in the bacterial composition at the phylum level even after 20 d post feeding (dpf); however, the abundance of Intestinimonas and Barnesiella decreased in both RPS treatment groups compared to the CON group. Consumption of 5% RPS increased the abundance of Roseburia (p<0.05) and decreased the abundance of Clostridium (p<0.01) and Mediterraneibacter (p< 0.05). In contrast, consumption of 10% RPS increased the abundance of Olsenella (p<0.05) and decreased the abundance of Campylobacter (p<0.05), Kineothrix (p<0.05), Paraprevotella (p<0.05), and Vallitalea (p<0.05). Additionally, acetate (p<0.01), butyrate (p<0.05), valerate (p = 0.01), and total SCFAs (p = 0.01) were upregulated in the RPS5 treatment group. CONCLUSION: Feeding 5% RPS altered bacterial community composition and promoted gut health in weaned piglets. Thus, resistant starch as a feed additive may prevent diarrhea in piglets during weaning.

Synthesis of fluorescent organic nano-dots and their application as efficient color conversion layers.

Efficient conversion of light from short wavelengths to longer wavelengths using color conversion layers (CCLs) underpins the successful operation of numerous contemporary display and lighting technologies. Inorganic quantum dots, based on CdSe or InP, for example, have received much attention in this context, however, suffer from instability and toxic cadmium or phosphine chemistry. Organic nanoparticles (NPs), though less often studied, are capable of very competitive performance, including outstanding stability and water-processability. Surfactants, which are critical in stabilizing many types of nano-structures, have not yet been used extensively in organic NPs. Here we show the utility of surfactants in the synthesis and processing of organic NPs by thoroughly characterizing the effect of ionic and non-ionic surfactants on the properties of fluorescent organic NPs. Using this information, we identify surfactant processing conditions that result in nearly 100 % conversion of organic fluorophores into sub-micrometer particles, or nano-dots, with outstanding performance as CCLs. Such water dispersions are environmentally benign and efficiently convert light. They can be used for a range of fluorophores covering a full spectral gamut, with excellent color purity, including full-width at half-maximum (FWHM) values as low as 21 nm. Compared to inorganic (InP) reference CCLs, the organic nano-dot based CCLs show superior color conversion efficiency and substantially improved long-term stability.

Erratum to: Evaluation of adenosine triphosphate testing for on-farm cleanliness monitoring compared to microbiological testing in an empty pig farrowing unit.

[This corrects the article DOI: 10.5187/jast.2020.62.5.682.].

Acceptor-Donor-Acceptor-Type Orange-Red Thermally Activated Delayed Fluorescence Materials Realizing External Quantum Efficiency Over 30% with Low Efficiency Roll-Off.

Two new orange-red thermally activated delayed fluorescence (TADF) materials, PzTDBA and PzDBA, are reported. These materials are designed based on the acceptor-donor-acceptor (A-D-A) configuration, containing rigid boron acceptors and dihydrophenazine donor moieties. These materials exhibit a small ΔEST of 0.05-0.06 eV, photoluminescence quantum yield (PLQY) as high as near unity, and short delayed exciton lifetime (τd ) of less than 2.63 µs in 5 wt% doped film. Further, these materials show a high reverse intersystem crossing rate (krisc ) on the order of 106 s-1 . The TADF devices fabricated with 5 wt% PzTDBA and PzDBA as emitting dopants show maximum EQE of 30.3% and 21.8% with extremely low roll-off of 3.6% and 3.2% at 1000 cd m-2 and electroluminescence (EL) maxima at 576 nm and 595 nm, respectively. The low roll-off character of these materials is analyzed by using a roll-off model and the exciton annihilation quenching rates are found to be suppressed by the fast krisc and short delayed exciton lifetime. These devices show operating device lifetimes (LT50 ) of 159 and 193 h at 1000 cd m-2 for PzTDBA and PzDBA, respectively. The high efficiency and low roll-off of these materials are attributed to the good electronic properties originatng from the A-D-A molecular configuration.

A New BODIPY Material for Pure Color and Long Lifetime Red Hyperfluorescence Organic Light-Emitting Diode.

A red fluorescent material, 1,3,7,9-tetrakis(4-(tert-butyl)phenyl)-5,5-difluoro-10-(2-methoxyphenyl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborinine (4tBuMB), as an emitting dopant in a thermally activated delayed fluorescence (TADF) sensitized hyperfluorescence organic light-emitting diode (HFOLED) is reported. The 4tBuMB shows a high photoluminescence quantum yield (PLQY) of 99% with an emission maximum at 620 nm and a full width at half-maximum (fwhm) of 31 nm in solution. Further, it shows a deep lowest unoccupied molecular orbital (LUMO) of 3.83 eV. Thus, two TADF materials, 4CzIPN and 4CzTPN, as sensitizing hosts, are selected on the basis of a suitable LUMO level and spectrum overlap with 4tBuMB. The fabricated HFOLED device with 4CzTPN as a sensitizing host and 4tBuMB as an emitting dopant shows a maximum external quantum efficiency (EQE), an emission maximum, an fwhm, and CIE coordinates of 19.4%, 617 nm, 44 nm, and (0.64, 0.36), respectively. The electroluminance performances of the 4CzTPN sensitized device are higher than those of the 4CzIPN-based device, which is attributed to a higher Förster resonance energy transfer (FRET) rate and reduced intersystem crossing/reverse intersystem crossing (ISC/RISC) cycles of the former. Also, the 4CzTPN-based HF device shows a longer device lifetime (LT90) of 954 h than the 4CzIPN-baed device (LT90 of 57 h) at 3000 cd m-2. The higher device stability is due to the higher bond dissociation energies (BDEs) of 4CzTPN and 4tBuMB than that of 4CzIPN.

Evaluation of adenosine triphosphate testing for on-farm cleanliness monitoring compared to microbiological testing in an empty pig farrowing unit.

Careful cleaning and disinfection of pigpens is essential to prevent disease spread and avoid the resultant economic losses. Hygiene in pigpens is generally evaluated by visual monitoring supplemented with bacteriological monitoring, which includes counting the total aerobic bacteria (TAB) and/or fecal indicator bacteria (FIB). However, these methods present drawbacks such as time and labor requirements. As adenosine triphosphate (ATP) is ubiquitous in all living organisms including microorganisms, this study aimed to directly compare the results of microbial assessment and ATP quantification, and to suggest possible detailed application methods of the ATP test for hygiene evaluation in pigpens of a farrowing unit. Before and after standard cleaning procedures, samples were collected from the floor corner, floor center, and feeding trough of four pigpens at different time points. No FIB were detected and both the TAB and ATP levels were significantly decreased in the floor center area after cleaning. FIB were continuously detected after cleaning and disinfection of the floor corners, and there was no significant ATP level reduction. The feeding trough did not show any significant difference in these values before and after cleaning, indicating insufficient cleaning of this area. The levels of TAB and ATP after cleaning were significantly correlated and the average ATP value was significantly lower in the absence of FIB than in their presence. In the absence of standard references, a more thorough hygiene management could be achieved evenly by supplementing cleaning or disinfection based on the lowest ATP results obtained at the cleanest test site, which in the present study was the floor center. Overall, these results indicate that the on-farm ATP test can be used to determine the cleanliness status, in addition to visual inspection, as an alternative to laboratory culture-based testing for the presence of microorganisms.

Comparative Analysis between Short Stem and Conventional Femoral Stem in Patients with Osteonecrosis of Femoral Head: Metha Stem and Excia Stem.

OBJECTIVE: To compare the intraoperative, radiological, and clinical short-term outcomes of cementless total hip arthroplasties (THA) using a short stem (SS) and a conventional femoral stem (CS) in a randomized prospective control study. METHODS: From June 2011 to October 2017, patients who underwent cementless THA for idiopathic osteonecrosis of the femoral head were recruited. Patients had a minimum 2 years of follow-up after the operation. The patients were divided into two groups: those who underwent THA using an SS and those who underwent THA using a CS. SS were used in 34 patients (41 hips) and CS were used in 41 patients (45 hips). In both groups, the same cup was used in all cases, and the mean follow-up periods were 63 (26-101) months in the SS and 64 (26-101) months in the CS groups. Intraoperative, clinical, and radiological evaluations were performed for the two groups. RESULTS: There was no difference in the demographics of the two groups. There was one patient with a proximal femoral crack in the SS group and one with a distal femoral crack in the CS group. Clinically, the mean Harris hip score was improved in both groups at 2-year follow-up. Radiographically endosteal osseointegrations were found in 40 of 41 cases in the SS group and in 44 of 45 cases in the CS group. There was one case of dislocation in each group. In the SS group, the acetabular cup was changed and repositioned 7 months after the initial operation. Stem loosening, infection, ceramic breakage, and varus/valgus change were not observed. There was a statistically significant lower stress shielding effect in the SS group. There were no differences in vertical/parallel offset and leg length discrepancy. CONCLUSION: The intraoperative, radiological, and clinical evaluations in both groups showed good outcomes and there was no statistically significant difference between the two groups.

Effectiveness of Hip Arthroscopy Performed Simultaneously before Open Reduction and Internal Fixation for Acetabular Fracture and Fracture-dislocation of the Hip.

PURPOSE: This study is performed to evaluate the usefulness of arthroscopic surgery prior to open reduction and fixation surgery to treat acetabular fractures and hip fractures-dislocation. MATERIALS AND METHODS: From January 2010 to March 2014, a total of 54 patients with acetabular fractures or hip fractures with dislocation were treated arthroscopically via fracture surface before open reduction and fixation (group 1, n=11), and without hip arthroscopy prior to open reduction and fixation (group 2; n=43). Clinical results were evaluated using Harris hip score (HHS) and visual analogue scale (VAS) pain scores. RESULTS: The mean age of patients is 43.2 years and there are 10 males and 1 females in group 1. The mean follow-up period is 15 months. The acetabular status of each case was assessed arthroscopically. Bone fragment was performed in 6 cases, and ligamentum teres shrinkage in 1 case. At the final follow up, the mean HHS and VAS pain scores were 78.6 and 2.18, respectively. During follow up, one case of osteoarthritis and one case of heterotopic ossification were identified. At the final follow up, the mean HHS and VAS pain scores were 77.5 and 2.23, respectively. In group 2, oteoarthritis and ectopic ossification were observed in 4 and 1 cases, respectively. CONCLUSION: No differences were observed in the clinical outcomes of patients with acetabular fracture or hip fracture-dislocation when treated with or without arthroscopic surgery before open reduction and fixation. However, arthroscopy is thought to be useful for evaluating the joint cartilage surface and fracture fragments more accurately.

Prevalence of severe fever with thrombocytopenia syndrome virus in black goats (Capra hircus coreanae) in the Republic of Korea.

Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging tick-borne pathogen in China, Japan, and the Republic of Korea (ROK). The aim of this study was to investigate the prevalence of SFTSV antigens and anti-SFTSV antibodies in black goats (Capra hircus coreanae) throughout the ROK. Sera were collected from 737 black goats in nine provinces in the ROK. Eighteen of 737 (2.4%) goat sera were positive for SFTSV on one-step reverse transcription nested polymerase chain reaction. The amplified 346-bp S segments of SFTSV sequences were classified into three genotypes (BG1, BG2, and BG3), and were included in the Japanese clade rather than the Chinese clade, based on phylogenetic analysis. Forty-three of 624 (6.9%) serum samples were seropositive for anti-SFTSV antibodies on enzyme-linked immunosorbent assay analysis. This study is the first to examine the molecular prevalence of SFTSV in goats and the first to perform serological detection of anti-SFTSV antibodies in livestock in the ROK. Moreover, the results indicate that SFTSV is widely distributed in goats and that additional monitoring for SFTSV is needed in livestock in the ROK.

Cytological endometritis in dairy cows: diagnostic threshold, risk factors, and impact on reproductive performance.

We determined the threshold proportion of polymorphonuclear leukocytes (PMNs) for a diagnosis of cytological endometritis (CEM), the risk factors for this condition, and its impact on reproductive performance in dairy cows. Uterine cytology was performed on 407 Holstein cows 4 weeks postpartum to determine the proportions of endometrial cells and PMNs. A receiver operator characteristics curve was used to determine the threshold above which the PMN proportion affected the likelihood of cows conceiving by 200 days postpartum. The optimal threshold was ≥ 14% PMN (sensitivity, 31.3%; specificity, 81.7%; p ＜ 0.05). The farm identity, retained placenta (odds ratio [OR] = 1.87), and septicemic metritis (OR = 3.07) were risk factors for CEM (p ＜ 0.05). Cows with CEM were less likely to resume cyclicity (OR = 0.58) and to conceive by 200 days postpartum (hazard ratio = 0.58). Cows with CEM tended (p ＜ 0.1) to be less likely to become pregnant after their first insemination (OR = 0.65) and to require a greater number of inseminations per conception (2.3 vs. 2.2). In conclusion, a PMN threshold of 14% defined the presence of CEM at 4 weeks postpartum. The farm, retained placenta, and septicemic metritis were risk factors for CEM, which reduces subsequent reproductive performance.

Effect of two treatment protocols for ketosis on the resolution, postpartum health, milk yield, and reproductive outcomes of dairy cows.

We determined the effect of ketosis treatment with propylene glycol (PG) or PG plus l-carnitine and methionine (Metabolase®, Fatro, Bologna, Italy) on the resolution, postpartum health, milk yield, and reproductive performances of dairy cows. Blood from 475 Holstein cows was collected weekly until 4 weeks after calving to measure blood ß-hydroxybutyrate (BHBA) concentrations. Cows with blood BHBA concentration ≥1.2 mmol/L were diagnosed with ketosis and were enrolled. One hundred and fifty cows diagnosed with ketosis were randomly assigned to three treatment groups (Day 0): (1) PG (300 g, PO) for 3 days (PG group, n = 50), (2) PG (300 g, PO) plus l-carnitine (1.25 g) plus methionine (5 g, IV) for 3 days (PG + CM group, n = 50), and (3) no treatment (control group, n = 50). On Day 3, blood was collected to evaluate whether the ketosis had resolved. Cows in the PG and PG + CM groups with blood BHBA ≥1.2 mmol/L were retreated for an additional 2 days, and then blood BHBA concentration was evaluated on Days 5 and 10. Blood glucose and haptoglobin concentrations, rumen fill score (RFS), and body condition score (BCS) were measured on Days 0, 3, 5, and 10. Postpartum complications, milk yield during the first 2 months, and reproductive outcomes were evaluated. The probability of resolution from ketosis was higher (P < 0.05) in the PG + CM group than in the control group on Days 3, 5, and 10 (odds ratio: 2.6-6.3). Blood BHBA in the PG + CM group was lower (P < 0.05) than that of the control group on Days 3 and 5, whereas blood glucose in the PG + CM group was higher (P < 0.05) than that of the control group on Days 3 and 5. RFS in the PG and PG + CM groups was higher than that of the control group on Day 10 (P < 0.01), while BCS loss from Day 0-10 in the control group was higher than those of the PG and PG + CM groups (P < 0.05). Milk yields on the 30th and 60th days postpartum were higher in the PG + CM group than the control and PG groups (P < 0.05). Postpartum complications and intervals between calving and first postpartum insemination or pregnancy did not differ among the groups (P > 0.05). In conclusion, treatment of dairy cows with PG plus l-carnitine and methionine improved the chances of resolution of ketosis and increased milk yield, while affecting neither the incidence of postpartum complications nor reproductive performance.

Rumen fermentation and digestibility of spent mushroom ( Pleurotus ostreatus ) substrate inoculated with Lactobacillus brevis for Hanwoo steers / Fermentación ruminal y digestibilidad de la cama de champiñón (Pleurotus ostreatus) desechada inoculada con Lactobacillus brevis en novillos Hanwoo / Fermentação ruminal e digestibilidade de resíduo de substrato de cogumelo (Pleurotus ostreatus) inoculado com Lactobacillus brevis en novilhos Hanwoo

Abstract Background: Ensiling spent mushroom substrate (SMS) generally increases its nutrient digestibility and quality. Objective: To determine the feed quality and digestibility of SMS from Pleurotus ostreatus (SMSP) inoculated with lactic acid bacteria (LAB: Lactobacillus brevis) in Hanwoo steers. Methods: Ruminal disappearance of SMSP and inoculated SMSP (ISMSP) were evaluated in three rumen-fistulated Hanwoo steers (408 ± 13.0 Kg body weight). Further, three healthy Hanwoo steers (336 ± 69.0 Kg body weight) were randomly allotted to one of three dietary treatments (control: 25% straw, 75% concentrates; treatments: 25% straw, 60% concentrates, and 15% of either SMSP or ISMSP) in a 3×3 Latin square design. Results: The chemical composition of the ISMSP diet did not differ from that of the control or the SMSP diets. In the ISMSP diet, the rate of decrease of pH of ruminal fluid and the increase in storage period was greater than with the SMSP diet. Ruminal disappearance of dry matter, crude protein, neutral detergent fiber, and acid detergent fiber were slightly higher in steers fed ISMSP than those fed SMSP. Furthermore, the degradation rate and effective degradability of crude protein was greater in the ISMSP diet than in the SMSP diet. Effective ruminal fermentation characteristics and total nutrients digestibility were not affected by SMSP nor ISMSP diet. Conclusion: The SMSP and ISMSP diets could replace formulated concentrate without adverse effects and be a cost-effective feed for Hanwoo steers. Furthermore, LAB inoculation improved the SMSP preservation.

Resumen Antecedentes: El ensilado de cama de champiñón desechada (SMS) generalmente aumenta la digestibilidad y la calidad de sus nutrientes. Objetivo: Determinar la calidad del alimento y digestibilidad del SMS a partir de Pleurotus ostreatus (SMSP) inoculado con bacterias ácido-lácticas (LAB: Lactobacillus brevis) en bueyes Hanwoo. Métodos: La desaparición ruminal del SMSP y ISMSP (SMSP inoculado) fue evaluada en tres bueyes Hanwoo fistulados en el rumen (408 ± 13,0 Kg peso corporal). Igualmente, tres bueyes Hanwoo sanos (336 ± 69,0 Kg peso corporal) fueron asignados al azar a uno de los tres tratamientos dietéticos (control: 25% de heno, 75% de concentrados; tratamientos: 25% de heno, 60% de concentrados y 15% de SMSPo ISMSP) en un diseño cuadrado latino 3×3. Resultados: La composición química de la dieta ISMSP no difirió de la del control o de la dieta SMSP. En la dieta ISMSP, la tasa de disminución del pH del fluido ruminal y el incremento del tiempo de almacenamiento fueron mayores que los de la dieta SMSP. La desaparición ruminal de la materia seca, proteína cruda, fibra detergente neutra y la fibra detergente ácida fue ligeramente superior en los bueyes alimentados con el ISMSPque en aquellos alimentados con SMSP. Además, la tasa de degradación y la degradabilidad efectiva de la proteína cruda fueron mayores en la dieta ISMSPque en la dieta SMSP. Las características de la fermentación ruminal efectiva y la digestibilidad total de nutrientes no fueron afectadas por la dieta SMSP ni por la de ISMSP. Conclusión: Las dietas SMSPe ISMSP podrían reemplazar el concentrado formulado sin efectos adversos y ser una alimentación económica para los bueyes Hanwoo. Además, la inoculación con LAB mejoró la conservación del SMSP.

Resumo Antecedentes: Ensilagem de resíduo de substrato de cogumelo (SMS) geralmente aumenta a digestibilidade e a qualidade dos nutrientes. Objetivo: Determinar a qualidade da alimentação e digestibilidade do RSC apartir do Pleurotus ostreatus (SMSP) inoculado com bactérias de ácido láctico (LAB: Lactobacillus brevis) nos novilhos Hanwoo. Métodos: Desaparecimento ruminal do SMSP e ISMSP (SMSP inoculado) foram avaliados em três novilhos Hanwoo fistulados no rúmen (408 ± 13.0 Kg peso corporal). Além disso, três novilhos Hanwoo (336 ± 69.0 Kg peso corporal) foram aleatoriamente distribuídos para um dos três tratamentos dietéticos (controle: 25% palha, 75% concentrado; tratamentos: 25% palha, 60% concentrado, e 15% ambos SMSP e ISMSP) em um quadrado Latino de 3×3. Resultados: A composição química da dieta ISMSP não diferiu do controle ou das dietas SMSP. Na dieta ISMSP, a taxa de diminuição do pH do fluido ruminal e o aumento no período de armazenamento foram melhores do que com a dieta SMSP. O desaparecimento ruminal de matéria seca, proteína bruta, fibra em detergente neutro, e fibra detergente ácido foi ligeiramente superior em novilhos alimentados com ISMSP, do que aqueles alimentados com SMSP. Além disso, a taxa de degradação e degradabilidade da proteína bruta foi maior na dieta ISMSP, do que na dieta SMSP. As características efetivas de fermentação ruminal e a digestibilidade total de nutrientes não foram afetadas pela dieta SMSP, nem pela dieta ISMSP. Conclusão: Dietas com SMSP e ISMSP podem ser utilizados em substituição ao concentrado formulado sem causar efeitos adversos e ser um alimento rentável em novilhos Hanwoo. Além disso, a inoculação com LAB melhorou a qualidade conservante da SMSP.

Green fluorescence protein-based content-mixing assay of SNARE-driven membrane fusion.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediate intracellular membrane fusion by forming a ternary SNARE complex. A minimalist approach utilizing proteoliposomes with reconstituted SNARE proteins yielded a wealth of information pinpointing the molecular mechanism of SNARE-mediated fusion and its regulation by accessory proteins. Two important attributes of a membrane fusion are lipid-mixing and the formation of an aqueous passage between apposing membranes. These two attributes are typically observed by using various fluorescent dyes. Currently available in vitro assay systems for observing fusion pore opening have several weaknesses such as cargo-bleeding, incomplete removal of unencapsulated dyes, and inadequate information regarding the size of the fusion pore, limiting measurements of the final stage of membrane fusion. In the present study, we used a biotinylated green fluorescence protein and streptavidin conjugated with Dylight 594 (DyStrp) as a Föster resonance energy transfer (FRET) donor and acceptor, respectively. This FRET pair encapsulated in each v-vesicle containing synaptobrevin and t-vesicle containing a binary acceptor complex of syntaxin 1a and synaptosomal-associated protein 25 revealed the opening of a large fusion pore of more than 5 nm, without the unwanted signals from unencapsulated dyes or leakage. This system enabled determination of the stoichiometry of the merging vesicles because the FRET efficiency of the FRET pair depended on the molar ratio between dyes. Here, we report a robust and informative assay for SNARE-mediated fusion pore opening.