Ammonia‐oxidizing Archaea in laboratory‐scale activated sludge systems for wastewater of low‐ or high‐ammonium concentration

Ammonia‐oxidizing bacteria (AOB) is known as ammonia‐oxidizer in wastewater treatment systems. However, ammonia‐oxidizing Archaea (AOA) is found from various environments, including wastewater treatment systems. In this study, to investigate the relationships between AOA population and ammonia concentration, AOA was monitored in two laboratory‐scale reactors treating artificial wastewater of different ammonium concentrations by denaturing gradient gel electrophoresis targeting ammonia monooxygenase genes. At day 60 of the operation, AOA populations dominant in each reactor differed, suggesting the importance of influent ammonia concentration in dominant AOA selection.     

Prothrombin time and activated partial thromboplastin time using a point‐of‐care analyser (Abaxis VSpro®) in Bennett's wallabies (Macropus rufogriseus)

There are few reports of coagulation times in marsupial species. Blood samples collected from 14 Bennett's wallabies (Macropus rufogriseus) under anaesthesia during routine health assessments were analysed for prothrombin time (PT) and activated partial thromboplastin time (aPTT) using a point‐of‐care analyser (POC) (Abaxis VSPro®). The wallabies had an aPTT mean of 78.09 s and median of 78.1 s. The PT for all wallabies was greater than 35 s, exceeding the longest time measured on the POC. Although PT was significantly longer, aPTT was similar to the manufacturer's domestic canine reference range.     

Effects of water‐misting sprays with forced ventilation on post mortem glycolysis,AMP‐activated protein kinase and meat quality of broilers after transport during summer

Effects of water‐misting sprays with forced ventilation on post mortem glycolysis, adenosine monophosphate‐activated protein kinase (AMPK) and meat quality of broilers after transport during summer were investigated in the present paper. A total of 105 mixed‐sex Arbor Acres broilers were divided into three treatment groups: (i) 45 min transport without rest (T); (ii) 45 min transport with 1 h rest (TR); and (iii) 45 min transport with 15 min water‐misting sprays with forced ventilation and 45 min rest (TWFR). Each treatment consisted of five replicates with seven birds each. The results indicated that the water‐misting sprays with forced ventilation could mitigate the stress caused by transport under high temperature conditions during summer, which reduced the energy depletion in post mortem Pectoralis major (PM) muscle. This resulted in a higher energy status compared to the T group, which would decrease the expression of phosphorylation of AMPK (p‐AMPK). Furthermore, decreased the expression of p‐AMPK then slowed down the rate of glycolysis in post mortem PM muscle during the early post mortem period, which in turn lessened the negative effects caused by transport on meat quality. In conclusion, water‐misting sprays with forced ventilation may be a better method to control the incidence of the pale, soft and exudative meat in broilers.     

Low‐Molecular‐Weight Heparin Dosage in Newborn Foals

Background: Heparin is used in humans as prophylaxis of hypercoagulable states and disseminated intravascular coagulation (DIC). However, babies need a higher heparin dose than do adults. Septic neonate foals are at high risk of hypercoagulable state and DIC, and there is limited objective information about heparin dose for equine neonates. Objective: To assess whether neonate foals require higher dosages of low‐molecular‐weight heparin (LMWH) than adults. Animals: Eighteen healthy and 11 septic neonate foals. Methods: Experimental and clinical studies. Firstly, healthy foals were randomly distributed in 2 groups, 1 receiving 50 IU/kg SC of dalteparin and the 2nd group receiving 100 IU/kg SC of dalteparin, once daily for 3 days. Blood samples were collected before and 3, 6, 27, and 51 hours after the 1st LMWH administration. Plasma antifactor‐Xa activity was measured, together with hemostatic and hematologic parameters used to assess the risk of bleeding. Subsequently, septic foals were treated blindly either with placebo (saline) or 100 IU/kg of dalteparin for 3 days. Plasma antifactor‐Xa activity and other hemostatic parameters were determined before and after treatment. Results: Plasma antifactor‐Xa activity in healthy foals was below prophylactic activity when using the adult dosage (50 IU/kg), whereas prophylactic activities were achieved when using the double dosage (100 IU/kg). No hemorrhagic events and erythrocyte‐related complications were observed with either dosage. In the clinical study, only 4/6 septic foals had plasma antifactor‐Xa activity adequate for prophylaxis. Conclusions and Clinical Importance: Equine neonates require higher dosages of LMWH compared with adults to reach prophylactic heparinemia.     

Combining ultrasound and lactobacilli treatment for high‐fat‐diet‐induced obesity in mice

Chronic systemic lipopolysaccharide‐induced inflammation can cause obesity. In animal experiments, lactobacilli have been shown to inhibit obesity by modifying the gut microbiota, controlling inflammation and influencing the associated gene expression. A previous study found that high‐fat‐diet‐induced (HFD) obesity was suppressed by lactobacilli ingestion in rats via the inhibition of parasympathetic nerve activity. This study explored the combined use of lactobacilli ingestion and ultrasound (US) to control body weight and body fat deposition in HFD mice over an 8‐week experimental period. Male C57BL/6J mice received an HFD during treatment and were randomly divided into four groups: (i) control group (H), (ii) lactobacilli alone (HB), (iii) US alone (HU) and (iv) lactobacilli combined with US (HUB). The US was targeted at the inguinal portion of the epididymal fat pad on the right side. At the 8th week, body weight had decreased significantly in the HUB group (15.56 ± 1.18%, mean ± SD) group compared with the HU (26.63 ± 0.96%) and H (32.62 ± 5.03%) groups (p < 0.05). High‐resolution microcomputed tomography (micro‐CT) scans revealed that the reduction in total body fat volume was significantly greater in the HUB group (69%) than in the other two experimental groups (HB, 52%; HU, 37%; p < 0.05). The reductions in the thickness of the subcutaneous epididymal fat pads were significantly greater in the HUB group (final thickness: 340 ± 7 μm) than in the H (final thickness: 1150 ± 21 μm), HB (final thickness: 1060 ± 18 μm) and HU (final thickness: 370 ± 5 μm) groups (all p < 0.05). Combination therapy with lactobacilli and US appears to enhance the reduction in body weight, total and local body fat deposition, adipocyte size and plasma lipid levels over an 8‐week period over that achieved with lactobacilli or US alone in HFD mice. These results indicate that US treatment alone can reduce hyperlipidemia in HFD mice.     

N‐Acetyl‐d‐galactosamine prevents soya bean agglutinin‐induced intestinal barrier dysfunction in intestinal porcine epithelial cells

Soya bean agglutinin (SBA) is a glycoprotein and the main anti‐nutritional component in most soya bean feedstuffs. It is mainly a non‐fibre carbohydrate‐based protein and represents about 10% of soya bean‐based anti‐nutritional effects. In this study, we sought to determine the effects of N‐Acetyl‐D‐galactosamine (GalNAc or D‐GalNAc) on the damage induced by SBA on the membrane permeability and tight junction proteins of piglet intestinal epithelium (IPEC‐J2) cells. The IPEC‐J2 cells were pre‐cultured with 0, 0.125 × 10^?4, 0.25 × 10^?4, 0.5 × 10^?4, 1.0 × 10^?4 and 2.0 × 10^?4 mmol/L GalNAc at different time period (1, 2, 4 and 8 hr) before being exposed to 0.5 mg/ml SBA for 24 hr. The results indicate that pre‐incubation with GalNAc mitigates the mechanical barrier injury as reflected by a significant increase in trans‐epithelial electric resistance (TEER) value and a decrease in alkaline phosphatase (ALP) activity in cell culture medium pre‐treated with GalNAc before incubation with SBA as both indicate a reduction in cellular membrane permeability. In addition, mRNA levels of the tight junction proteins occludin and claudin‐3 were lower in the SBA‐treated groups without pre‐treatment with GalNAc. The mRNA expression of occludin was reduced by 17.3% and claudin‐3 by 42% (p < 0.01). Moreover, the corresponding protein expression levels were lowered by 17.8% and 43.5% (p < 0.05) respectively. However, in the GalNAc pre‐treated groups, occludin and claudin‐3 mRNAs were reduced by 1.6% (p > 0.05) and 2.7% (p < 0.01), respectively, while the corresponding proteins were reduced by 4.3% and 7.2% (p < 0.05). In conclusion, GalNAc may prevent the effect of SBA on membrane permeability and tight junction proteins on IPEC‐J2s.     

The membrane‐type estrogen receptor G‐protein‐coupled estrogen receptor suppresses lipopolysaccharide‐induced interleukin 6 via inhibition of nuclear factor‐kappa B pathway in murine macrophage cells

The female sex hormone estrogen exerts anti‐inflammatory effects. The G‐protein‐coupled estrogen receptor (GPER) has been recently identified as a novel membrane‐type estrogen receptor that can mediate non‐genomic estrogenic effects on many cell types. We previously demonstrated that GPER inhibits tumor necrosis factor alpha‐induced expression of interleukin 6 (IL‐6) through repression of nuclear factor‐kappa B (NF‐κB) promoter activity using human breast cancer cells. Although several reports have indicated that GPER suppresses Toll‐like receptor‐induced inflammatory cytokine expression in macrophages, the molecular mechanisms of the inhibition of cytokine production via GPER remain poorly understood. In the present study, we examined GPER‐mediated inhibition of IL‐6 expression induced by lipopolysaccharide (LPS) stimulation in a mouse macrophage cell line. We found that the GPER agonist G‐1 inhibited LPS‐induced IL‐6 expression in macrophage cells, and this inhibition was due to the repression of NF‐κB promoter activity by GPER. G‐1 treatment also decreased the phosphorylation of inhibitor of κB kinases. Among the mitogen‐activated protein kinases, the phosphorylation of c‐jun N‐terminal kinase (JNK) was increased by G‐1. These findings delineate the novel mechanism of the inhibition of LPS‐induced IL‐6 through GPER‐activated JNK‐mediated negative regulation of the NF‐κB pathway in murine macrophage cells, which links anti‐inflammatory effects to estrogen.     

Analysis of glycosuria in okapi (Okapia johnstoni): Examination of urinary N‐acetyl‐β‐D‐glucosaminidase

We analyzed the urinary excretion of glucose and N‐acetyl‐β‐D‐glucosaminidase (NAG) in six okapis (Okapia johnstoni) in captivity to investigate the cause of their urinary sugar excretion. The urinary glucose‐positive okapi had significantly higher urinary NAG indices than the urinary glucose‐negative okapi. There was also a positive correlation between urinary glucose levels and urinary NAG indices. These results suggest that the proximal tubular function of the glycosuric okapi may have been obstructed, which impaired glucose reabsorption.     

Pharmacokinetics and bioavailability after intramuscular injection of the 5‐HT1A serotonin agonist R‐8‐hydroxy‐2‐(di‐n‐propylamino) tetralin (8‐OH‐DPAT) in domestic goats (Capra aegagrus hircus)

To determine the bioavailability and pharmacokinetic properties of the serotonin 5‐HT_1A receptor agonist R‐8‐OH‐DPAT in goats, and 0.1 mg kg^?1 R‐8‐OH‐DPAT hydrobromide was administered intramuscularly (i.m.) and intravenously (i.v.) to six goats in a two‐phase cross‐over design experiment. Venous blood samples were collected from the jugular vein 2, 5, 10, 15, 20, 30, 40 and 60 min following treatment and analysed by liquid chromatography tandem mass spectrometry. Bioavailability and pharmacokinetic parameters were determined by a one‐compartment analysis. Mean bioavailability of R‐8‐OH‐DPAT when injected i.m. was 66%. The mean volume of distribution in the central compartment was 1.47 L kg^?1. The mean plasma body clearance was 0.056 L kg^?1 min^?1. All goats injected i.v. and two of six goats injected i.m. showed signs of serotonin toxicity. In conclusion, R‐8‐OH‐DPAT is well absorbed following i.m. injection and the observed pharmacokinetics suggest that administration via dart is feasible. Administration of R‐8‐OH‐DPAT hydrobromide, at a dosage of 0.1 mg kg^?1, resulted in the observation of clinical signs of serotonin toxicity in the goats. It is suggested that dosages for the clinical use of the compound should be lower in order to achieve the desired clinical effect without causing serotonin toxicity.     

Non‐immunosuppressive FTY720‐derivative OSU‐2S mediates reactive oxygen species‐mediated cytotoxicity in canine B‐cell lymphoma

OSU‐2S is a FTY720 (Fingolimod) derivative that lacks immunosuppressive properties but exhibits strong anti‐tumour activity in several haematological and solid tumour models. We have recently shown OSU‐2S to mediate potent cytotoxicity in human mantle cell lymphoma cell lines and primary cells. We report here the pre‐clinical activity of OSU‐2S in spontaneous B‐cell lymphoma of dogs which shares many characteristics of human lymphoma. OSU‐2S mediated apoptosis in canine B‐cell lines and primary B‐cell lymphoma cells obtained from spontaneous lymphoma bearing dogs. OSU‐2S induced reactive oxygen species (ROS) in canine lymphoma cells and inhibition of ROS partially rescued OSU‐2S‐mediated cell death. These studies provide a rational basis for the use of spontaneous lymphoma in pet dogs as a preclinical large animal model for the development of OSU‐2S as small molecule for treating people and dogs with lymphoma.     

Insulin‐independent actions of glucagon‐like peptide‐1 in wethers

Insulin‐independent actions of glucagon‐like peptide‐1 (GLP‐1) are not yet clear in ruminants. Four Suffolk mature wethers (60.0 ± 6.7 kg body weight (BW)) were intravenously infused with insulin (0.5 mU/kg BW/min; from 0 to 90 min) and GLP‐1 (0.5 μg/kg BW/min; from 60 to 150 min) with both hormones co‐administered from 60 to 90 min, in a repeated‐measure design under euglycemic clamp for 150 min, to investigate whether GLP‐1 has insulin‐independent actions. Jugular blood samples were taken at 15‐min intervals for plasma hormones and metabolites analysis. Compared to baseline concentrations (at 0 min), insulin infusion decreased (P < 0.05) plasma concentrations of glucagon, non‐esterified fatty acids (NEFA), lactate, nonessential amino acids (NEAA), branched‐chain amino acids (BCAA), total amino acids (TAA) and urea nitrogen (UN). Insulin plus GLP‐1 infusion induced a greater increase (P < 0.05) in plasma concentrations of insulin and triglyceride (TG), but decreased (P < 0.05) glucagon, total cholesterol (T‐Cho), NEAA and UN plasma concentrations. GLP‐1 infusion increased (P < 0.05) NEFA, β‐hydroxybutyrate and TG, but decreased (P < 0.05) glucagon, T‐Cho, NEAA, BCAA and UN plasma concentrations. In conclusion, GLP‐1 exerts extrapancreatic roles in ruminants not only insulin‐independent but probably, in contrast to non‐ruminants, antagonistic to insulin effects.     

The localization and function of p38α mitogen‐activated protein kinase in rat oocytes

P38α mitogen‐activated protein kinase (MAPK), which is a member of the canonical MAPK family, is activated in response to various extracellular stresses and plays a role in multiple cellular processes. In this study, we investigated the expression, subcellular localization and functional roles of p38α MAPK during the meiotic maturation of rat oocytes. We found that p38α MAPK phosphorylation (p‐p38α MAPK, indicative of p38α MAPK activation) was low at the germinal vesicle (GV) stage, increased 3 hr after germinal vesicle breakdown (GVBD) and maintained its maximum at metaphase I (MI) or metaphase II (MII). The p‐p38α MAPK mainly accumulated in the GV and had no obvious expression in the nucleus. From GVBD to MII, p‐p38α MAPK was distributed in the cytoplasm around either the chromosomes or the spindle. We used SB203580, an inhibitor of p38α MAPK, to investigate the possible functional role of p38α MAPK during rat oocyte meiotic maturation. Treatment of GV stage oocytes with 20 μM SB203580 blocked p‐p38α MAPK activity, and the spindles appeared abnormal. Additionally, the rate of GVBD after 3 hr of culture with 20 μM SB203580 (58.8%) was significantly inhibited compared with the control (82.5%, p < .05), and the polar body extrusion rate after 12 hr of culture with SB203580 was also significantly decreased compared with the control (40.1% vs 73.3%, p < .05). Taken together, these data indicate that p38α MAPK may play a vital role in rat oocyte meiotic maturation.