L-lactate Concentration Research Articles

Lactate drives cellular DNA repair capacity: Role of lactate and related short-chain fatty acids in cervical cancer chemoresistance and viral infection

The characteristic feature of a cancer microenvironment is the presence of a highly elevated concentration of L-lactate in the tumor niche. The lactate-rich environment is also maintained by commensal mucosal microbiota, which has immense potential for affecting cancer cells through its receptoric and epigenetic modes of action. Some of these lactate activities might be associated with the failure of anticancer therapy as a consequence of the drug resistance acquired by cancer cells. Upregulation of cellular DNA repair capacity and enhanced drug efflux are the most important cellular mechanisms that account for ineffective radiotherapy and drug-based therapies. Here, we present the recent scientific knowledge on the role of the HCA1 receptor for lactate and lactate intrinsic activity as an HDAC inhibitor in the development of an anticancer therapy-resistant tumor phenotype, with special focus on cervical cancer cells. In addition, a recent study highlighted the viable role of interactions between mammalian cells and microorganisms in the female reproductive tract and demonstrated an interesting mechanism regulating the efficacy of retroviral transduction through lactate-driven modulation of DNA-PKcs cellular localization. To date, very few studies have focused on the mechanisms of lactate-driven enhancement of DNA repair and upregulation of particular multidrug-resistance proteins in cancer cells with respect to their intracellular regulatory mechanisms triggered by lactate. This review presents the main achievements in the field of lactate impact on cell biology that may promote undesirable alterations in cancer physiology and mitigate retroviral infections.

Interactive effects of food deprivation state and hypoxia on the respiratory responses of postprandial rock crabs, Cancer irroratus.

Under the background of climate change, increasing attention has focused on the effects of ocean deoxygenation on marine organisms. However, few studies address the effects of different food deprivation states on hypoxia tolerance. We therefore investigated the metabolic responses of the Atlantic rock crab, Cancer irroratus (starved 28-35 days, fasted 3-5 days and recently fed). Starved-crab exhibited the lowest critical oxygen saturation (Scrit), while fed-crab had the highest Scrit. The fed-crab maintained an elevated postprandial oxygen consumption (MO2) even below the Scrit of fasted-crab indicating reserved aerobic scopes for critical activities in severe hypoxia. Following feeding, hypoxia (50% and 20% oxygen saturation, SO2) retarded the specific dynamic action resulting in lower peak MO2 and longer duration. The starved-crab exhibited a lower peak MO2, prolonged duration and higher energy expenditure than fasted-crab after feeding. The decline in arterial PO2 was most pronounced below the Scrit for both fasted- and starved-crab. The higher hemocyanin concentration ([Hc]) of fasted-crab (than starved-crab) suggested they had improved oxygen transport capacity, but hypoxia did not increase [Hc] during the 72-h experiment. Following feeding, the fasted-crab significantly increased L-lactate concentration ([L-lactate]) in 20% SO2, which was not observed in starved-crab. These results suggest starvation may trigger a cross-tolerance to hypoxia. Because crabs can undergo long periods of food deprivation in their natural environment, future studies should consider how this may affect their ability to deal with environmental perturbations.

PSXIII-2 Effect of Rest, Post-Rest Transport Duration, and Conditioning on Performance and Physiological Welfare Indicators of Beef Calves

Abstract The aim of this study was to assess the effects of conditioning, rest, and post-rest transport duration on welfare indicators of 6-7 mo old beef calves. A total of 328 weaned calves (237 ± 29.7 kg BW) were randomly assigned to a 2 × 2 × 2 factorial design: conditioning, conditioned (C) or non-conditioned (N); rest, 0 (R0) or 8 (R8) h, and post-rest transport, 4 (T4) or 15 (T15) h. Calves were sampled prior to and after 20h of transport, prior to and after the additional 4 or 15-h transport, and at 1, 2, 3, 5, 14, and 28 d after transport ended. Data were analyzed using the GLIMMIX procedure of SAS. Fixed effects were conditioning, transport and time nested within rest period, while random effects were animal and pen. For R0-T4 calves, the mean L-lactate concentrations were greater than R8-T4 calves on d 1 and 2 (p-values = 0.02) while, for R0 calves, mean ADG was greater than R8 calves 14 to 28 d after transport (p-values < 0.01). For R8-T4 calves 1 week after transport, mean ADG was greater and mean WBC counts were less than R8-T15 calves on d 5 (p-values < 0.01). For N calves, overall mean haptoglobin, creatine kinase, serum amyloid-A, and non-esterified fatty acids were greater than C calves (p-values ≤ 0.05). Overall, few and inconsistent differences were observed for rest, where rest improved L-lactate but affected ADG. Few differences were observed for transport, where shorter transport durations after the rest improved ADG and WBC counts. In general, N calves had greater physiological indicators of reduced welfare than C calves. Based on these findings, the best way to improve calf welfare during and after transport is to condition them prior to transport.

PM2.5 promotes NSCLC carcinogenesis through translationally and transcriptionally activating DLAT-mediated glycolysis reprograming

BackgroundAirborne fine particulate matter (PM2.5) has been associated with lung cancer development and progression in never smokers. However, the molecular mechanisms underlying PM2.5-induced lung cancer remain largely unknown. The aim of this study was to explore the mechanisms by which PM2.5 regulated the carcinogenesis of non-small cell lung cancer (NSCLC).MethodsParalleled ribosome sequencing (Ribo-seq) and RNA sequencing (RNA-seq) were performed to identify PM2.5-associated genes for further study. Quantitative real time-PCR (qRT-PCR), Western blot, and immunohistochemistry (IHC) were used to determine mRNA and protein expression levels in tissues and cells. The biological roles of PM2.5 and PM2.5-dysregulated gene were assessed by gain- and loss-of-function experiments, biochemical analyses, and Seahorse XF glycolysis stress assays. Human tissue microarray analysis and 18F-FDG PET/CT scans in patients with NSCLC were used to verify the experimental findings. Polysome fractionation experiments, chromatin immunoprecipitation (ChIP), and dual-luciferase reporter assay were implemented to explore the molecular mechanisms.ResultsWe found that PM2.5 induced a translation shift towards glycolysis pathway genes and increased glycolysis metabolism, as evidenced by increased L-lactate and pyruvate concentrations or higher extracellular acidification rate (ECAR) in vitro and in vivo. Particularly, PM2.5 enhanced the expression of glycolytic gene DLAT, which promoted glycolysis but suppressed acetyl-CoA production and enhanced the malignancy of NSCLC cells. Clinically, high expression of DLAT was positively associated with tumor size, poorer prognosis, and SUVmax values of 18F-FDG-PET/CT scans in patients with NSCLC. Mechanistically, PM2.5 activated eIF4E, consequently up-regulating the expression level of DLAT in polysomes. PM2.5 also stimulated transcription factor Sp1, which further augmented transcription activity of DLAT promoter.ConclusionsThis study demonstrated that PM2.5-activated overexpression of DLAT and enhancement in glycolysis metabolism contributed to the tumorigenesis of NSCLC, suggesting that DLAT-associated pathway may be a therapeutic target for NSCLC.

Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased \u03b22 adrenergic agonist

Metabolic reprogramming contributes to oncogenesis, tumor growth, and treatment resistance in pancreatic ductal adenocarcinoma (PDAC). Here we report the effects of (R,S′)-4′-methoxy-1-naphthylfenoterol (MNF), a GPR55 antagonist and biased β2-adrenergic receptor (β2-AR) agonist on cellular signaling implicated in proliferation and metabolism in PDAC cells. The relative contribution of GPR55 and β2-AR in (R,S′)-MNF signaling was explored further in PANC-1 cells. Moreover, the effect of (R,S′)-MNF on tumor growth was determined in a PANC-1 mouse xenograft model. PANC-1 cells treated with (R,S′)-MNF showed marked attenuation in GPR55 signal transduction and function combined with increased β2-AR/Gαs/adenylyl cyclase/PKA signaling, both of which contributing to lower MEK/ERK, PI3K/AKT and YAP/TAZ signaling. (R,S′)-MNF administration significantly reduced PANC-1 tumor growth and circulating l-lactate concentrations. Global metabolic profiling of (R,S′)-MNF-treated tumor tissues revealed decreased glycolytic metabolism, with a shift towards normoxic processes, attenuated glutamate metabolism, and increased levels of ophthalmic acid and its precursor, 2-aminobutyric acid, indicative of elevated oxidative stress. Transcriptomics and immunoblot analyses indicated the downregulation of gene and protein expression of HIF-1α and c-Myc, key initiators of metabolic reprogramming in PDAC. (R,S′)-MNF treatment decreased HIF-1α and c-Myc expression, attenuated glycolysis, shifted fatty acid metabolism towards β-oxidation, and suppressed de novo pyrimidine biosynthesis in PANC-1 tumors. The results indicate a potential benefit of combined GPR55 antagonism and biased β2-AR agonism in PDAC therapy associated with the deprogramming of altered cellular metabolism.

Performance, health, bacterial metabolites and intestinal histomorphology in does and growing rabbits fed diets with increasing lignocellulose-to-cellulose proportions

ABSTRACT Adequate levels of dietary fibre can reduce the risk of intestinal health disorders in rabbits after weaning. Therefore, it was the aim of the study to investigate the impact of different lignocellulose and cellulose addition to rabbits’ diets on performance and intestinal traits in lactating does and weaned rabbits. A total of 60 rabbit does (4.41 ± 0.45 kg body weight) were fed isonitrogenous diets with lignocellulose and cellulose inclusion rates [%] of 0/8, 2/6, 4/4, 6/2 and 8/0, resulting in dietary lignin-to-cellulose ratios of 0.26, 0.30, 0.32, 0.34 and 0.40, respectively. The diets were fed from 4 d ante partum (a.p.) until day 28 post partum (p.p. weaning). After weaning, 40 young rabbits from each feeding group were fed the same diets for 12 d, when 10 rabbits per group were sacrificed. Body weight gain (BWG), feed intake and feed conversion ratio were obtained. Gastrointestinal content and gut tissue were obtained from the growing rabbits and pH, short-chain fatty acids (SCFA) concentration and gut morphology were investigated. Digestive disorders were scored, and the sanitary risk index was calculated. Does fed the highest lignocellulose addition increased their feed intake [g · kg BW−1 · d−1] in the first two weeks p.p. (p < 0.001) and increased BWG (p < 0.001) but without impact on their offsprings’ performance. Weaned rabbits fed the highest lignocellulose inclusion showed a reduced feed intake and increased sanitary risk index. Additionally, a higher number of rabbits showed intestinal disorders for a short time (1–2 d, p < 0.001). In the colon content, a higher pH and a lower dry matter content were observed in this group. Caecal contents of 40-d old rabbits showed lower total SCFA, acetate and propionate and higher i-valerate and L-lactate concentrations after feeding diets with high lignocellulose contents. The diet did not influence the acetate-to-propionate ratio. No histomorphological changes were observed in the jejunum, while crypt depth and width were reduced in the caecum of weaned rabbits fed diets with lignocellulose addition. It can be concluded that different proportions of lignocellulose to cellulose addition to rabbits’ diets have an impact on health and performance with differences between does and weaned rabbits. While a high lignocellulose inclusion proved beneficial for does, for weaned rabbits, a lower addition should be preferred.

Shredding of grass-clover before ensiling: Effects on feed intake, digestibility, and methane production in dairy cows

Shredding is a type of processing that mechanically treats forage in order to separate plant tissues apart and rupture the plant cell. This experiment aimed at investigating the effects of shredding grass-clover harvested at two developmental stages on feed intake, digestibility, and gas production. The grass-clover was harvested either at an early (May 14; ERL) or a late (May 29; LAT) developmental stage. Within each developmental stage, grass-clover was mown, wilted, and either baled and wrapped (CON) or shredded, baled, and wrapped (SHR). The four combinations of ensiled grass-clover (ERL-CON, ERL-SHR, LAT-CON, and LAT-SHR) was fed to four rumen, duodenum, and ileum cannulated primiparous Danish Holstein cows in a 4 × 4 Latin square design with four periods of 21 d duration. The silage was offered for ad libitum intake. Silage density was higher (P < 0.01) for SHR compared to CON. Silage concentrations of L-lactate and acetate were higher (P = 0.01 and P < 0.01, respectively) and the silage concentration of butyrate was lower (P < 0.01) for SHR compared to CON, whereas pH was lower (P = 0.02), which indicated silage quality improved from shredding. The dry matter (DM) intake (DMI), milk yield, and rumen digestibility of neutral detergent fibre (aNDFom) were not affected by shredding, whereas feeding SHR compared to CON resulted in lower (P = 0.05) total tract digestibility of aNDFom (714 vs. 727 g/kg) and lower (P = 0.04) methane (CH4) production (60 vs. 66 L CH4/kg organic matter (OM) digested in the rumen). Compared to CON, SHR had a higher (P = 0.02) proportion of butyrate in rumen fluid indicating that shredding had some effect on the dynamics of rumen fermentation. However, the effective degradability of CP and aNDFom in the rumen determined in situ showed no effect of shredding. Rumination and total chewing time were lower (P = 0.03 and P = 0.05, respectively) and the concentration of protein in the milk was higher (P = 0.02), when shredding LAT compared to ERL. Furthermore, concentrations of CP and purines in rumen microbes were lower (P = 0.04 and P = 0.01, respectively), when cows were fed LAT compared to ERL. In conclusion, shredding improved silage fermentation quality, reduced digestibility of aNDFom without affecting the digestibility of other nutrients, and reduced methane production per kg of rumen digested OM. Furthermore, we observed no benefits from shredding grass-clover harvested at a late compared to early developmental stage, except for increased concentration of protein in milk and reduced time spent for chewing during rumination and total chewing.

Additive prognostic value of preoperative plasma glucose concentrations in calves undergoing abdominal surgery

Surgical abdominal emergencies in calves are associated with a guarded prognosis, especially if neonates are affected. Because hypoglycemia has been associated with sepsis and endotoxemia, this study aimed to assess the prognostic relevance of preoperative plasma glucose concentrations (GLUC) in calves requiring surgery for an acute abdominal disorder. For this purpose, data from retrospective and prospective case series were analyzed, consisting of 586 and 83 hospitalized calves, respectively. The outcomes of calves were evaluated until hospital discharge (both study populations) and for 3 mo following discharge by a phone call to the farmer (prospective study population). For the retrospective study population, the overall survival rate was 31.2%. Calves with a negative outcome (NO) had significantly lower median GLUC (4.3 mmol/L) than calves with a positive outcome (PO; 5.0 mmol/L). The survival rates of calves with GLUC <2.4 mmol/L and 2.4 to 3.1 mmol/L were 3.6 and 8.3%, respectively. The inclusion of GLUC improved a previous prognostic model based on plasma l-lactate concentration and age. The resulting analyses indicated that NO was associated with low age (<7 d), hyper-l-lactatemia (>8.84 mmol/L), and GLUC <4.4 mmol/L (age 7–20 d) and <3.3 mmol/L (age ≥21 d), respectively. The area under the receiver operating characteristic curve of this model was 0.79 (95% confidence interval: 0.76–0.83) and the resulting sensitivity and specificity for NO at the optimal probability cut-point of 0.69 were 66.7 and 85.8%, respectively. For the prospective study population, the established model had sensitivity and specificity for predicting NO after 3 mo (proportion 24%) of 61.9 and 85%, respectively. In both study populations, hypoglycemia was significantly associated with intraoperative evidence of a septic process within the abdominal cavity. The present analyses show that hypoglycemia was highly indicative of a poor prognosis and serious intraoperative findings such as peritonitis. Determination of GLUC should therefore be part of the diagnostic work-up in calves suffering from an acute abdominal emergency.

1 Effect of Conditioning, Rest, and Post-rest Transport Duration on Welfare Indicators of Beef Cattle Transported by Road

Abstract The aim of this study was to assess the effects of conditioning, rest, and post-rest transport duration on welfare indicators of 6–7 mo old beef calves. Three hundred and twenty-eight weaned calves (237 ± 29.7 kg BW) were randomly assigned to a 2 × 2 × 2 nested factorial design: conditioning, conditioned (C) or non-conditioned (N); rest, 0 (R0) or 8 (R8) h, and post-rest transport, 4 (T4) or 15 (T15) h. Calves were sampled prior to the first loading (L1), after 20h of transport, prior to and after the additional 4 or 15-h transport, and at 1, 2, 3, 5, 14, and 28 d after transport ended. Data were analyzed using the GLIMMIX procedure of SAS. Fixed effects included conditioning, transport and time nested within rest period, while random effects included animal and pen. Greater shrink (P &amp;lt; 0.01) was observed in C than N calves after the initial 20-h transport. The N calves had greater (P &amp;lt; 0.01) ADG than C calves between L1 and d 5, while C had greater (P &amp;lt; 0.01) ADG than N calves between 14 and 28 d. L-lactate concentrations and flight speed were greater (P ≤ 0.05) in C than N calves between L1 and d 5. The R8-T4 calves had greater (P &amp;lt; 0.01) ADG than R8-T15 calves between L1 and d 5. The R0-T4 calves had greater (P = 0.02) L-lactate concentrations than R0-T15 and R8-T4 calves on d 1. The R0 calves had greater (P &amp;lt; 0.01) ADG than R8 calves between 14 and 28 d. Preliminary results show physiological, behavioral, and performance differences across treatments, however, additional indicators are required to accurately assess the effect of conditioning, rest, and post-rest transport durations on calf welfare.

Assessment of preoperative and postoperative l-lactatemia and clinical outcomes in goats undergoing tube cystostomy: A retrospective study of 34 cases (2015-2020).

BackgroundUrolithiasis in small ruminants has a poor long‐term prognosis, and long‐term clinical outcomes are variable and unpredictable.ObjectivesTo assess the accuracy of preoperative and postoperative blood l‐lactate concentrations in predicting a negative outcome in goats undergoing tube cystostomy.AnimalsThirty‐four male goats undergoing tube cystostomy.MethodsRetrospective study. Medical records of goats undergoing tube cystostomy from 2015 to 2020 were reviewed. Clinical variables recorded included signalment, procedures before surgery, urolith location and type, duration of hospitalization, and heart rate. PCV, plasma total protein, potassium, preoperative and postoperative blood l‐lactate concentrations, preoperative and postoperative creatinine concentrations, and relative changes in blood l‐lactate and creatinine concentrations over time were measured using heparinized blood. A negative outcome was defined as death or euthanasia from urolithiasis complications at 6 months after discharge. Negative outcomes as a function of independent clinical variables were evaluated using χ2 or Fisher's exact tests, and multivariate logistic regression. P < .05 was considered significant.ResultsMedian (95% confidence interval) preoperative, postoperative, and the relative change over time of blood l‐lactate concentrations were 3.3 mmol/L (2.2, 4.8), 1.0 mmol/L (0.7, 1.3), and 0.4 mmol/L (−3.5, 3.2), respectively. Preoperative (P = 1), postoperative (P = .14), and the relative change over time (P = .63) of blood l‐lactate concentrations were not significant predictors of a negative outcome. Furthermore, all other clinical variables measured were not significant predictors of a negative outcome (P > .05).Conclusions and Clinical ImportanceVeterinarians should advise clients that clinical outcomes after tube cystostomy in goats are likely unpredictable.

Systemic implications of metallic foreign body syndrome in dairy cattle

This study aimed to evaluate the systemic implications of 37 cattle with traumatic reticulitis, evaluating clinical, laboratory, ultrasonographic and necroscopic changes. A clinical, laboratory and ultrasonographic examination was performed, and the animals were distributed in reticuloperitonitis (GI; n=21) and reticulopericarditis (GII; n=16) traumatic, based on necropsy findings. Blood samples were taken for haematological tests, serum total protein, albumin, globulin, gamma glutamyltransferase (GGT), aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH), creatinine, urea, cortisol, creatine kinase (CK), creatine kinase-MB (CK-MB), cardiac troponin-I (cTn-I), plasma glucose and L-lactate. The abdominocentesis was productive in eight GI (n=8) and seven GII (n=7) animals, allowing the evaluation of physical, cytological and biochemical characteristics of the peritoneal fluid as a total protein, albumin, GLDH, AST, GGT, glucose and L-lactate. Changes in behavior, appetite, dehydration and temperature were observed, most expressive in GII. Hematology showed neutrophilic leukocytosis with regenerative left shift and hyperfibrinogenemia in both groups. Increased globulin, L-lactate concentration and serum GGT, GLDH, CK and CK-MB activity were observed, as well as significant elevation of cTnI (p=0.0190) in GII. In the peritoneal fluid exudate was observed in both groups and a higher concentration of L-lactate in relation to plasma. Ultrasound revealed retocular, cardiac, hepatic and splenic abnormalities. The anatomopathological lesions confirmed the ultrasound findings of both groups. The understanding of the syndrome helps in the diagnosis, as well as the adoption of preventive measures, minimizing the economic impact caused to the dairy cattle breeding.

L-lactic acidosis confers insensitivity to PKC inhibitors by competing foruptake via monocarboxylate transporters.

Targeting protein kinase C (PKC) family was found to repress the migration and resistance of non-small cell lung cancer cells to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs). However, none of the PKC inhibitors has been approved for anticancer therapy yet due to the limited efficacy in clinical trials, and the underlying mechanisms remain unclear. l-lactic acidosis, a common condition comprising high l-lactate concentration and acidic pH in the tumor microenvironment, has been known to induce tumor metastasis and drug resistance. In this study, l-lactic acid was found to reverse the inhibitory effects of pan-PKC inhibitors GO6983 on PKC activity, cell migration, and EGFR-TKI resistance, but these effects were not affected by the modulators of lactate receptor GPR81. Interestingly, blockade of lactate transporters, monocarboxylate transporter-1 and -4 (MCT1 and MCT4), attenuated the intracellular level of GO6983, and its inhibitory effect on PKC activity, suggesting that lactic acid promotes the resistance to PKC inhibitors by competing for the uptake through these transporters rather than by activating its receptor, GPR81. Our findings explain the underlying mechanisms of the limited response of PKC inhibitors in clinical trials.

Comparison of selected serum biochemistry measurements between the Nova Prime Plus VET, Nova pHOx Ultra, and Beckman Coulter AU680 analyzers in dogs.

Blood gas chemistry analyzers typically produce results faster and use smaller sample volumes than reference chemistry analyzers. However, results may not be comparable between blood gas chemistry analyzers and reference chemistry analyzers or between different models of blood gas chemistry analyzers. This could suggest the use of separate reference intervals and, thus, has implications when making clinical decisions. We aimed to perform method comparison studies to evaluate selected canine serum biochemical values obtained using the Nova Stat Profile Prime Plus VET (Prime Plus VET), Stat Profile Nova pHOx Ultra (Ultra), and Beckman Coulter AU680 (Beckman) analyzers. We hypothesized that the three analyzers would be identical within inherent imprecision. Jugular venous blood samples were collected from 103 endurance-trained sled dogs, and serum was harvested and stored for analysis. Results for serum chloride, potassium, sodium, creatinine, and urea nitrogen concentrations obtained from the Prime Plus VET and Ultra analyzers were compared with results from the Beckman analyzer, which was considered to be a reference method. Results for serum chloride, potassium, sodium, creatinine, urea nitrogen, and L-lactate concentrations obtained from the Prime Plus VET and Ultra analyzers were compared. Passing-Bablok regression and Bland-Altman plots were used for method comparison. Significant (P<0.05) constant or proportional bias was found for many analytes for all three method comparison studies. Due to the presence of statistically significant differences between all three analyzers that may be clinically relevant, it is recommended that reference intervals be created for new blood gas analyzers, even when similar methodologies are used.

Specific inhibition of Streptococcus bovis by endolysin LyJH307 supplementation shifts the rumen microbiota and metabolic pathways related to carbohydrate metabolism

BackgroundEndolysins, 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.ResultsLyJH307 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.ConclusionsLyJH307 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.

Participation of L-Lactate and Its Receptor HCAR1/GPR81 in Neurovisual Development.

During the development of the retina and the nervous system, high levels of energy are required by the axons of retinal ganglion cells (RGCs) to grow towards their brain targets. This energy demand leads to an increase of glycolysis and L-lactate concentrations in the retina. L-lactate is known to be the endogenous ligand of the GPR81 receptor. However, the role of L-lactate and its receptor in the development of the nervous system has not been studied in depth. In the present study, we used immunohistochemistry to show that GPR81 is localized in different retinal layers during development, but is predominantly expressed in the RGC of the adult rodent. Treatment of retinal explants with L-lactate or the exogenous GPR81 agonist 3,5-DHBA altered RGC growth cone (GC) morphology (increasing in size and number of filopodia) and promoted RGC axon growth. These GPR81-mediated modifications of GC morphology and axon growth were mediated by protein kinases A and C, but were absent in explants from gpr81−/− transgenic mice. Living gpr81−/− mice showed a decrease in ipsilateral projections of RGCs to the dorsal lateral geniculate nucleus (dLGN). In conclusion, present results suggest that L-lactate and its receptor GPR81 play an important role in the development of the visual nervous system.

Daily alternation of the dietary starch level in Holstein dairy cows

The aim of this study was to investigate the effect of controlled daily alternations in dietary starch level on changes in rumen environment, blood, urine, and milk metabolites of dairy cows. Six multiparous mid-lactation Holstein cows were used in a replicated 3 × 3 Latin square design with 14-d periods and 3 alternating levels of dietary starch as treatments. Each 14-d period consisted of a 7-d baseline period and 7-d alternating period where diets alternated day to day. During the baseline period, all cows were fed a control diet containing 21% starch (dry matter basis). During the alternating period, the control diet was replaced with 1 of the 3 experimental diets on d 8, 10, 12, and 14. The 3 experimental diets contained 28% (low), 35% (medium), and 42% (high) starch (dry matter basis). At d 7 (baseline), 8 (ALT1), and 14 (ALT4) of each period, rumen fluid, blood, urine, and quarter milk (i.e., back right quarter) samples were collected at -0.5, 1, 2.5, 4, 5.5, and 7 h relative to morning feeding (0800 h). No differences were observed in dry matter intake, milk yield, and milk chemical composition. Rumen medial pH was lower in the high alternation level compared with the low or medium alternation levels at ALT1 but did not differ among starch alternation levels at ALT4. Similarly, the difference between rumen pH in medial and ventral contents was reduced at ALT1 with high alternation level but was not affected at ALT4. Total volatile fatty acid (VFA) concentrations were higher in the rumen medial fluid of the high alternation level at 7 h relative to morning feeding compared with those from the low and medium alternation levels. Similarly, total VFA concentrations constantly increased and were the highest in the ventral rumen fluid at 7 h relative to morning feeding, although no differences were detected among starch alternation levels. In both rumen medial and ventral fluids, the high alternation level showed higher propionate and lower acetate proportions compared with low and medium alternation levels. No differences in blood pH were detected among starch alternation levels. However, glucose concentrations tended to be higher in cows from the high alternation level. l-Lactate concentrations in blood were higher in ALT1 than in ALT4 but were not affected by the starch alternation level. In urine, no differences in pH or l-lactate concentrations were detected among alternation levels (i.e., low, medium, and high). Similarly, no differences in milk pH were detected among alternation levels. According to these results, it seems that the daily dietary starch alternation from 21% up to 42% (dry matter basis) is able to affect the ruminal fluid, especially during the first alternation. However, these changes in rumen fluid did not cause any effect on the variables measured in blood, urine, or milk. This study indicates that cows can cope with day-to-day alternations in type of rumen fermentable organic matter; however, longer-term effects on performance and health should be addressed in future studies.

Risk factors for laminitis and nonsurvival in acute colitis: Retrospective study of 85 hospitalized horses (2011-2019).

BackgroundAcute colitis is a serious cause of morbidity and death in horses. Recent studies have compared clinical features of coronavirus and salmonellosis, but no study has compared clinical features of enteric salmonellosis, coronavirus, and neorickettsiosis.Hypothesis/ObjectivesTo identify risk factors for laminitis and nonsurvival to discharge in horses with enteric salmonellosis, coronavirus, or neorickettsiosis.AnimalsEighty‐five horses hospitalized for acute colitis from 2011 to 2019.MethodsRetrospective case series. Medical record review (2011‐2019) of adult (≥2 years) horses with colitis. Primary outcomes were laminitis and survival to discharge. Multivariable logistic regression was performed to assess association between variables and the development of laminitis. Stepwise Cox regression was performed to assess association between variables and survival.ResultsSeventeen of 85 (20%) horses developed laminitis during hospitalization. Neorickettsiosis cases (11/26, 42%) were more likely to develop laminitis than coronavirus (0/16, 0%) cases (odds ratio [OR] 24.48; 95% confidence interval [CI]: 1.33‐451.74, P = .03). There was no significant difference in laminitis between salmonellosis and neorickettsiosis cases (OR 0.27; 95% CI: 0.07‐1.07, P = .06). Admission heart rate (OR 1.08; 95% CI: 1.02‐1.15, P = .01), total solids (OR 0.17; 95% CI: 0.06‐0.54, P = .003), band neutrophils (OR 1248.47; 95% CI: 6.62‐235 540, P = .008), and bicarbonate concentration (OR 0.68; 95% CI: 0.5‐0.92, P = .01) were predictive of development of laminitis during hospitalization. Sixty‐three of 85 (74%) horses survived to discharge: 16/16 (100%) coronavirus cases, 17/26 (65%) neorickettsiosis cases, 14/20 (70%) salmonellosis cases, and 16/23 (70%) unknown cases. Packed cell volume (hazard ratio [HR] 1.17; 95% CI: 1.09‐1.26, P < .001), L‐lactate concentration (HR 1.33; 95% CI: 1.14‐1.55, P < .001), and development of laminitis (HR 7.07; 95% CI: 1.67‐29.95, P = .008) were retained in the final multivariable model for prediction of nonsurvival to discharge.Conclusion and Clinical ImportanceNonsurvival and laminitis rates were high, likely related to the presence of neorickettsiosis in the region.

Influence of endocrine disease on l-lactate concentrations in blood of ponies.

BackgroundBlood l‐lactate concentrations are higher in people with developing or established diabetes mellitus and insulin resistance.ObjectivesTo investigate whether blood l‐lactate concentrations are positively correlated with measures of insulin dysregulation (ID) or increased autumnal ACTH concentrations in ponies.AnimalsSystemically healthy client‐owned ponies (n = 101).MethodsProspective case‐control study. Blood samples were obtained from 101 clinically healthy ponies. Breed, weight, height, and subjective and objective measures of body condition were recorded. Blood l‐lactate, glucose, triglyceride, total adiponectin, and ACTH concentrations were measured and an oral sugar test (OST) was carried out. Correlations between blood l‐lactate and variables of endocrine health were determined.ResultsUsing a seasonal cutoff point of ACTH concentrations ≥47 pg/mL, 55 ponies had increased autumnal ACTH concentrations and 45 did not. Using a basal insulin concentration of >50 μiU/mL, 42 ponies were diagnosed with ID and 58 were not. Using a 60 minutes after OST cutoff point of >45 μiU/mL, 57 ponies had ID and 37 did not. Blood l‐lactate concentrations were significantly lower in obese (average body condition score ≥ 7/9) compared to nonobese ponies (0.6 mmol/L; range, 0.0‐1.9 mmol/L vs 0.8 mmol/L; range, 0.3‐2.7 mmol/L; P = .01). No other significant correlations were detected. No differences were detected between ponies with and without increased autumnal ACTH concentrations (0.7 mmol/L; range, 0.0‐2.7 mmol/L vs 0.7 mmol/L; range, 0.3‐1.8 mmol/L; P = .84) and with and without ID (0.7 mmol/L; range, 0.3‐2.7 mmol/L vs 0.8 mmol/L; range, 0.0‐1.6 mmol/L; P = .63).Conclusions and Clinical ImportanceResults do not support an effect of endocrine status on l‐lactate concentrations in blood of ponies.

Correlation between L-Lactate Concentrations in Beef Cattle, Obtained Using a Hand-Held Lactate Analyzer and a Lactate Assay Colorimetric Kit.

Simple SummaryLactate is a metabolite used in animal research as an indicator of muscle fatigue; therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ hand-held analyzer and a traditional lactate assay colorimetric kit. Blood samples were collected from 96 steers prior to loading and after 36 h of transportation, and prior to reloading and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ hand-held analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in the colorimetric analysis. Correlations were calculated to assess the strength of the relationship between the L-lactate concentrations measured between methods. The strength of the correlation and the level of statistical significance varied over the observed time points, while the correlation for the pooled data was weak. Based on the low strength of the correlation, the Lactate Scout+ analyzer is not a suitable alternative to a colorimetric assay for measuring L-lactate in transported cattle.Lactate is a product of anaerobic glycolysis, used in animal research as an indicator of muscle fatigue. Therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ analyzer and a traditional lactate assay colorimetric kit. Blood samples were collected by venipuncture from 96 steers (Black or Red Angus × Hereford/Simmental and Black or Red Angus × Charolais; 247 ± 38.2 kg BW) prior to loading (LO1) and after 36 h of transport, and prior to reloading and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in the colorimetric analysis. Pearson correlations were calculated to assess the strength of the relationship between the experimental methods for the quantification of L-lactate concentrations. The magnitude and direction of the correlation, and the level of statistical significance varied over the observed time points, ranging from r = −0.03 (p = 0.75; LO1) to r = 0.75 (p < 0.0001; d 3). The correlation for the pooled data was weak but statistically significant (r = 0.33, p < 0.0001). Based on the low magnitude of the correlation due to variability across sampling time points in this study, the Lactate Scout+ analyzer is not a suitable alternative to a lab-based assay (considered the gold standard) for measuring L-lactate in transported cattle.

Effects of handling during experimental procedures on stress indices in the green shore crab, Carcinus maenas (L)

ABSTRACT Stress due to handling is often an unavoidable feature of experimental investigations. In some cases, appropriate settling times are not considered, and as such, physiological responses caused by handling may become additive with those of experimental treatments. This study investigated the effect of different handling procedures on the acute physiological responses of green shore crab (Carcinus maenas). Handling, such as would occur during transport around a research facility or transfer during experimental procedure, was designated as light (10 min emersion) or severe (10 min emersion with shaking). Oxygen consumption (MO2) and haemolymph glucose and haemolymph L-lactate concentrations were elevated post-handling, the magnitude of the change related to the severity of handling stress. Glucose and L-lactate concentrations peaked within 1 h and returned to basal levels within 6 h, but MO2 remained elevated for 10 h, reflecting the additional energy required to oxidize L-lactate and replenish energy reserves. Differences between light and severe handling treatments showed that vibration (shaking) was a major contributor to the stress response, rather than the experimental emersion. This was confirmed in a second experiment where crabs were handled without emersion, and MO2 remained elevated for 14 h. In this experiment, the most pronounced increase in MO2 and metabolic parameters occurred in crabs that were physically touched and moved rapidly from the holding to experimental tanks. Here the touch, as well as vibration and visual stimuli, provoked a fight-flight response in the crabs. Stress responses were also evident in crabs gently transferred by containers. The fact that transferring crabs with no physical touching and minimal visual and vibrational stimuli still evoked a stress response, albeit less pronounced, supports a recommendation that crustaceans should be left to settle in the apparatus for at least 12 h after handling before experimental procedures are initiated.