An observational cohort study on antimicrobial usage on dairy farms in Quebec, Canada.

Quantification of antimicrobial usage (AMU) is crucial to measure the effect of intervention programs, to determine associations between usage and resistance, to compare populations, and for benchmarking purposes. The primary objective of the study was to describe quantitatively the AMU on Quebec dairy farms over 1 yr: (1) the total AMU, (2) the AMU per administration route (intramammary, injectable, oral, intrauterine), and (3) the AMU per antimicrobial class and according to the categorizations of Health Canada and the World Health Organization. The secondary objective was to assess the effect of several characteristics (herd size, level of milk production, and incidence rate of common infectious diseases) on AMU rate. The AMU data were obtained for 101 dairy farms randomly selected in 3 important Quebec dairy regions by collecting and recording all empty drug packaging and invoices for medicated feed (spring 2017 to spring 2018). The AMU rate was reported in number of Canadian defined course doses for cattle per 100 cow-years. The average herd size was 67 cows per farm, and 2/101 farms were certified organic. Overall, an estimated mean of 537 Canadian defined course doses for cattle/100 cow-years was observed. The intramammary route during lactation was the most frequently observed, followed, in decreasing order of usage, by oral route in the feed, intramammary route at drying-off, and injectable route. Oral (other than in animal feed) and intrauterine formulations were infrequently collected from the garbage cans. The 5 most frequently observed antimicrobial classes were, by decreasing order of usage, ionophores, penicillins, aminocoumarins, aminoglycosides, and polymyxins. Highest priority critically important antimicrobials as defined by the World Health Organization were mainly collected from intramammary formulations during lactation followed by injectable and drying-off intramammary formulations. The herd size was positively associated with the total AMU rate but not with the usage rate of highest priority critically important antimicrobials. Incidence of diseases along with preventive use of antimicrobials (drying-off and medicated feed with antimicrobials) explained 48% of the variance in total AMU rate.

Performance of beef heifers supplemented with sodium lasalocid.

This study was conducted on 78 13-month-old crossbred beef heifers that weighed 215 kg in Southern Rio Grande do Sul (RS) State, Brazil. We evaluated the performance of beef heifers that were reared in a pasture system that received a mineral supplement energy-type protein with added sodium lasalocid (LAS). The heifers were randomly and uniformly divided into 2 groups, with 39 animals in each group. One group of animals received a mineral supplement energy-type protein without sodium lasalocid (CON), and the other group received a mineral supplement energy-type protein with added LAS. The mean feed intake, the body weight (BW), the average daily gain (ADG), the body condition score (BCS), and ovarian cyclicity were recorded, and economic parameters were calculated. No differences in supplement intake were observed between the groups, which ensures adequate intake of the other components of the mineral mixture, which are part of the nutritional requirements for the production process. Similarly, no difference in the ADG was observed between treatments. We observed that the heifers in the LAS group had a higher BW gain (51 kg) that the CON heifers (40 kg; P < 0.05). In addition, LAS-supplemented heifers had a higher BCS (3.53) than CON heifers (3.38) at the end of the experiment (P < 0.05). The heifers supplemented with LAS had a higher profitability than the CON heifers, even with the higher cost of the supplement containing LAS; this effect was due to the higher live BW at the end of the study. We concluded that the administration of a mineral supplement energy-type protein with added LAS has beneficial effects on beef heifers in terms of production and economic feasibility.

Connectivity of the intracytoplasmic membrane of Rhodobacter sphaeroides: a functional approach.

The photosynthetic apparatus in the bacterium Rhodobacter sphaeroides is mostly present in intracytoplasmic membrane invaginations. It has long been debated whether these invaginations remain in topological continuity with the cytoplasmic membrane, or form isolated chromatophore vesicles. This issue is revisited here by functional approaches. The ionophore gramicidin was used as a probe of the relative size of the electro-osmotic units in isolated chromatophores, spheroplasts, or intact cells. The decay of the membrane potential was monitored from the electrochromic shift of carotenoids. The half-time of the decay induced by a single channel in intact cells was about 6 ms, thus three orders of magnitude slower than in isolated chromatophores. In spheroplasts obtained by lysis of the cell wall, the single channel decay was still slower (~23 ms) and the sensitivity toward the gramicidin concentration was enhanced 1,000-fold with respect to isolated chromatophores. These results indicate that the area of the functional membrane in cells or spheroplasts is about three orders of magnitude larger than that of isolated chromatophores. Intracytoplasmic vesicles, if present, could contribute to at most 10% of the photosynthetic apparatus in intact cells of Rba. sphaeroides. Similar conclusions were obtained from the effect of a ∆pH-induced diffusion potential in intact cells. This caused a large electrochromic response of carotenoids, of similar amplitude as the light-induced change, indicating that most of the system is sensitive to a pH change of the external medium. A single internal membrane and periplasmic space may offer significant advantages concerning renewal of the photosynthetic apparatus and reallocation of the components shared with other bioenergetic pathways.

Efficacy of in-feed preparations of an anticoccidial, multienzyme, prebiotic, probiotic, and herbal essential oil mixture in healthy and Eimeria spp.-infected broilers.

The efficacies of 5 widely used dietary supplements were investigated on performance indices, fecal oocyst excretion, lesion score, and intestinal tract measurements in healthy and Eimeria spp.-infected birds by using a comparative model. This study included 2,400 sexed Ross 308 broiler chicks that were equally divided in 2 groups: the infected group, experimentally infected with oocysts of mixed Eimeria spp. at 14 d of age, and the healthy controls. The birds in both groups were further divided equally into 6 groups, of which one was fed a basal diet and served as control without treatment and the other 5 served as experimental treatments. These 5 groups were fed 5 diets containing preparations of 60 mg/kg of anticoccidial salinomycin (SAL), 1 g/kg of multienzyme (ENZ), 1 g/kg of probiotic (PRO), 1 g/kg of prebiotic (PRE), and 40 mg/kg of an herbal essential oil mixture (EOM). Body weight gain and feed conversion ratio (FCR) showed significant improvement in the infected animals, which indicates that dietary supplemental regimens with SAL, ENZ, PRO, and PRE initiated in 1-d-old chicks reduced adverse effects after challenge with coccidiosis; however, chicks that were administered EOM failed to show such improvement. Uninfected chickens showed significant improvement in FCR with supplements SAL, PRE, and EOM, which signifies significant (P < 0.01) infection by supplement interactions for BW gain and FCR. In the infected group, all of the supplements reduced the severity of coccidiosis lesions (P < 0.01) induced by mixed Eimeria spp. through the middle and lower regions of the small intestines, whereas supplementation with SAL or EOM alone was effective (P < 0.01) in reducing oocyst excretion compared with the control treatment. The data indicated that use of these subtherapeutically efficacious supplements (except EOM) in broiler production can lessen the depression in growth due to coccidial challenge.

The impact of different levels of functional oil supplementation in combination with salinomycin on growth performance and intestinal microbiota of broilers undergoing Eimeria challenge: An analysis of dynamics.

The effect of salinomycin sodium alone and in combination with functional oils on performance and microbiota of broiler infected Eimeria were evaluated. 512 broilers were randomly assigned to 4 treatments (8 replicates, 16 birds/pen): a Control group (any additives); Ionophore group: salinomycin supplementation at 66 ppm (SS66); Ionophore +0.075% Functional oil (FO) group (SS66 + FO supplementation at 750 ppm); and Ionophore +0.10% FO group (SS66 + FO supplementation at 1000 ppm). At 14 days of age, birds were gavaged with 1 mL of a saline solution containing sporulated oocysts of E. tenella, E. acervulina and E. maxima. Performance indices were measured weekly. At 28 days, intestinal content was collected for microbiota analysis. Broilers of Control group presented the worst performance indices. Broilers of Ionophore + FO (0.075% and 0.10%) groups exhibited a higher BW at 28 days of age. The supplementation of Ionophore +0.075% FO resulted in a higher relative proportion of Firmicutes and a lower proportion of Actinobacteria in the ileum-jejunum. Lactobacillaceae was the dominant family in the jejunal, and ileal microbiotas of broilers fed diets supplemented with Ionophore, Ionophore +0.075% FO and Ionophore +0.10% FO. The supplementation of ionophore yielded higher numbers of Lactobacillaceae, Enterobactereaceae and Cloritridiaceae in the cecal. Ionophore associated with FO controlled the Lactobacillaceae, Enterobactereaceae and Cloritridiaceae families present in the cecum. Therefore, the combination of salinomycin with functional oil showed synergistic effect on performance and modulation of intestinal microbiota of broilers challenged with Eimeria.

Effect of three polyether ionophores on pharmacokinetics of florfenicol in male broilers.

Drug-drug interactions (DDIs) may adversely affect the prevention and cure of diseases. The effects of three polyether ionophore antibiotics, salinomycin (SAL), monensin (MON), and maduramycin (MAD) on the pharmacokinetics of florfenicol (FFC) were investigated in broilers. The chickens were fed rations with or without SAL (60 mg/kg feeds), MON (120 mg/kg feeds), or MAD (5 mg/kg feeds) for 14 consecutive days. FFC was given to the chickens either intravenously (i.v.) or orally (p.o.) at a single dose of 30 mg/kg body weight. Blood samples were taken from each chicken at 0-24 h postadministration of FFC. The plasma concentration of FFC was detected by high-performance liquid chromatography. The plasma concentration of FFC decreased with i.v. or p.o. co-administration of SAL, MON, or MAD in broilers, implying occurrence of DDIs during the co-administration of FFC with these ionophores. Our findings suggest that more attention should be given to the use of FFC to treat bacterial infections in chickens supplemented with polyether ionophore antibiotics.

Effects of a xylanase and protease, individually or in combination, and an ionophore coccidiostat on performance, nutrient utilization, and intestinal morphology in broiler chickens fed a wheat-soybean meal-based diet.

The effects of 2 single exogenous and monocomponent feed enzymes, and their combination, and an ionophore coccidiostat on production performance, feed AME(n), nutrient utilization, and intestinal morphology were studied in broiler chickens. One-day-old unvaccinated and unsexed Ross 308 birds (n = 320) were kept in groups of 8 on wood shavings in pens raised from the floor and fed one of 5 experimental diets, replicated 8 times, for 36 d. Treatments were 1) a wheat-soybean meal-based feed with no added coccidiostats or exogenous enzymes (CON), 2) CON + ionophore coccidiostat (Narasin), 3) CON + xylanase (Ronozyme WX CT; XYL), 4) CON + serine protease (Ronozyme ProAct CT; PRO), or 5) CON + xylanase + serine protease (XYL+PRO). Enzymes were added on top in the feed formulation. Diets contained 0.5% TiO2 to facilitate estimations of total tract apparent nutrient utilization. Treatments had no effect on BW gain or feed intake, but feed conversion, apparent digestibility of starch and fat, and feed AME(n) were improved with all enzyme treatments. The relative length of the ileum was reduced with XYL+PRO. For all parameters measured, the effects of XYL+PRO were similar to when XYL and PRO were fed individually. Narasin had no effect on production performance or nutrient utilization but reduced the relative lengths of jejunum and ileum. Relative lengths and weights of duodenum and cecum were unaffected by treatments. In conclusion, the improved feed conversion with both a xylanase and a protease was reflected in increased nutrient utilization, but their combination was not superior to when supplied separately. Narasin did not affect performance or nutrient utilization but reduced the relative lengths of the jejunum and ileum.

The LC-MS/MS-Based Measurement of Isopimaric Acid in Rat Plasma and Application of Pharmacokinetics.

Isopimaric acid (IPA) exhibits a diverse array of pharmacological activities, having been shown to function as an antihypertensive, antitumor, antibacterial, and hypocholesterolemic agent. However, few studies of the pharmacokinetics of IPA have been performed to date, and such analyses are essential to explore the in vivo mechanisms governing the biological activity of this compound. As such, we herein designed a selective LC-MS approach capable of quantifying serum IPA levels in model rats using an Agilent HC-C18 column (250 mm × 4.6 mm, 5 µm) via isocratic elution with a mobile phase composed of methanol 0.5% formic acid (91 : 9, v/v) at a 1 mL/min flow rate. Ion monitoring at m/z 301.2 [M-H]- was used to quantify IPA levels in plasma samples from these rats, while internal standard (IS) levels were assessed at m/z 455.3 [M-H]-. After validation, this approach was employed to conduct a pharmacokinetic analysis of rats administered IPA via the oral (p.o. 50, 100, or 200 mg/kg) and intravenous (i.v. 5 mg/kg) routes. Analyses of noncompartmental pharmacokinetic parameters revealed that IPA underwent secondary absorption following oral administration to these animals, with the two tested oral doses (50 and 100 mg/kg) being associated with respective absolute bioavailability values of 11.9% and 17.5%. In summary, this study may provide a foundation for future efforts to explore the mechanistic basis for the pharmacological activity of IPA, offering insights to guide its subsequent clinical utilization.

Evaluation of potential carryover effects associated with limit feeding of gravid Holstein heifers.

Ninety-six Holstein heifers (400±6kg, 15.2±0.1 mo), including 9 with ruminal cannulas, were offered 1 of 3 diets for 180±8 d in a randomized replicated pen design. Dietary treatments included a control diet (C100) and 2 independent limit-fed (LF) diets. The LF diets included one offered at 85% of C100 intake (L85) without an ionophore, and a second containing an ionophore (325 mg/head per day of lasalocid) that was offered at 80% of C100 intake (L80+I). Heifers were evaluated for growth, rumen digesta volume, nutrient excretion, and subsequent lactation performance. Limit-fed heifers consumed less dry matter and neutral detergent fiber, and had greater respective average daily gains (0.96 or 0.89 vs. 0.81 kg/d), and lower feed:gain ratios (9.1 or 9.3 vs. 13.0 kg/kg) compared with heifers offered the C100 diet. No differences in rumen pH, NH(3)-N, or volatile fatty acid concentrations were observed between C100 and LF heifers. Rumen digesta volume, density, and weight were unaffected by LF, and feeding L85 or L80+I did not result in carryover effects for rumen digesta volume when these heifers were offered a common high-fiber diet immediately after the 180-d growth trial. At parturition, no differences were observed for dystocia index, calf body weight, or 7-d postpartum body weight between cows offered LF or C100 diets as heifers. Lactation body weight, dry matter intake, and feed efficiency of cows did not differ between treatments at 45 or 90 d in milk. Milk yield and milk components also were not different between cows that were offered C100 or LF diets as gravid heifers. At 45 d in milk, rumen digesta volume was greater (99.1 vs. 66.1L) for cows offered L85 compared with cows offered L80+I as gravid heifers, but this effect was not observed at 90 d in milk. Limit feeding of gravid Holstein heifers for 180 d did not result in any carryover effects during their first lactation for rumen digesta volume, dry matter intake, or milk yield.

A field study on the effects of dietary monensin on milk production and milk composition in dairy cows.

The objectives of this study were to quantify the effect of 16 ppm of dietary monensin on milk production and composition of dairy cows, and to investigate factors having a potential impact on this effect. Data were generated from a total of 3577 Holstein dairy cows (47 herds) in Quebec enrolled in a herd-level, randomized clinical trial investigating the effects of monensin supplementation. Milk production and composition data were collected from monthly dairy herd improvement (DHI) testing. Monensin increased milk production by 0.9 kg/cow/d in cows under 150 days in milk (DIM) (P < 0.05). Monensin decreased milk fat percentage by 0.18 percentage points during the whole lactation (P < 0.05). This decreasing effect was larger for component-fed cows (P < 0.05) and for cows being fed low levels of dietary physically effective particles (P < 0.05) when compared respectively to cows fed total mixed ration and cows fed high levels of dietary physically effective particles. The results of this study suggest that monensin influences milk production and milk composition of dairy cows, and that diet composition and feeding system influence those effects.

Ionophore Use and Toxicosis in Cattle.

Ionophores are a commonly used feed additive for animals and when used properly are safe. When feed mis-mixing occurs and an elevated dose of ionophore is given, a toxicosis can develop. Myocardial and skeletal muscles are the targets of a toxicosis. In many species there is a delay from the time of ingestion of a toxic dose in feed to when clinical signs occur. This makes it difficult to collect the feed in question that was at an elevated concentration. Cardiac troponins in serum can be used to make a diagnosis of an ionophore toxicosis.

Salinomycin inhibits proliferative vitreoretinopathy formation in a mouse model.

Proliferative vitreoretinopathy (PVR) is a progressive disease that develops in a subset of patients who undergo surgery for retinal detachment repair, and results in significant vision loss. PVR is characterized by the migration of retinal pigment epithelial (RPE) cells into the vitreous cavity, where they undergo epithelial-to-mesenchymal transition and form contractile membranes within the vitreous and along the retina, resulting in recurrent retinal detachments. Currently, surgical intervention is the only treatment for PVR and there are no pharmacological agents that effectively inhibit or prevent PVR formation. Here, we show that a single intravitreal injection of the polyether ionophore salinomycin (SNC) effectively inhibits the formation of PVR in a mouse model with no evidence of retinal toxicity. After 4 weeks, fundus photography and optical coherence tomography (OCT) demonstrated development of mean PVR grade of 3.5 (SD: 1.3) in mouse eyes injected with RPE cells/DMSO (vehicle), compared to mean PVR grade of 1.6 (SD: 1.3) in eyes injected with RPE cells/SNC (p = 0.001). Additionally, immunohistochemistry analysis showed RPE cells/SNC treatment reduced both fibrotic (αSMA, FN1, Vim) and inflammatory (GFAP, CD3, CD20) markers compared to control RPE cells/DMSO treatment. Finally, qPCR analysis confirmed that Tgfß, Tnfα, Mcp1 (inflammatory/cytokine markers), and Fn1, Col1a1 and Acta2 (fibrotic markers) were significantly attenuated in the RPE cells/SNC group compared to RPE/DMSO control. These results suggest that SNC is a potential pharmacologic agent for the prevention of PVR in humans and warrants further investigation.

[The use of monensin premix in dairy cows: simple and essential steps for ensuring its proper use]. / L'utilisation du prémélange de monensin chez les vaches laitières: un suivi simple et essentiel pour s'assurer d'une utilisation adéquate.

The use of monensin premix in dairy cows: Simple and essential steps for ensuring its proper use. Dietary monensin, containing monensin sodium as active ingredient, is frequently used on dairy farms in Canada. Although the use of monensin is safe, some overdose situations have been reported following consumption of higher than recommended doses. A regular monitoring of bulk tank milk fat percentage should be performed to ensure quick detection of a potential overdose situation. Diarrhea and sudden drop in dry matter intake are other potential clinical signs of monensin overdose. Quick detection of such cases will allow rapid correction of the situation.(Translated by the authors).

PIKfyve accelerates phagosome acidification through activation of TRPML1 while arrests aberrant vacuolation independent of the Ca2+ channel.

PIKfyve phosphorylates PtdIns(3)P to PtdIns(3, 5)P2. One of the best characterized effector downstream of PtdIns(3, 5)P2 is a lysosomal Ca2+ channel, TRPML1. Although it has been reported that TRPML1 is involved in phagosome-lysosome fusion, the relevance of the Ca2+ channel in phagosome acidification has been denied. In this article, however, we demonstrated that the phagosome acidification was dependent on TRPML1. Based on the classical idea that Fluorescein isothiocyanate (FITC)-fluorescence is highly sensitive to acidic pH, we could estimate the phagosome acidification by time laps imaging. FITC-zymosan fluorescence that was engulfed by macrophages, decreased immediately after the uptake while the extinction of FITC-zymosan fluorescence was delayed in PIKfyve-deficient cells. The acidification arrest was completely rescued in the presence of Ca2+ ionophore A23187. Cells treated with a PIKfyve inhibitor, apilimod, also showed delayed phagosome acidification but were rescued by the overexpression of TRPML1. Additionally, TRPML1 agonist, ML-SA1 was effective to acidify the phagosome in PIKfyve-deficient cells. Another phenotype observed in PIKfyve-deficient cells is vacuole formation. Unexpectedly, enlarged vacuole formation in PIKfyve-deficient cells was not rescued by Ca2+ or over expression of TRPML1. It is likely that the acidification and vacuolation arrest is bifurcating downstream of PIKfyve.

A zwitterionic near-infrared fluorophore for real-time ureter identification during laparoscopic abdominopelvic surgery.

Iatrogenic injury of the ureters is a feared complication of abdominal surgery. Zwitterionic near-infrared fluorophores are molecules with geometrically-balanced, electrically-neutral surface charge, which leads to renal-exclusive clearance and ultralow non-specific background binding. Such molecules could solve the ureter mapping problem by providing real-time anatomic and functional imaging, even through intact peritoneum. Here we present the first-in-human experience of this chemical class, as well as the efficacy study in patients undergoing laparoscopic abdominopelvic surgery. The zwitterionic near-infrared fluorophore ZW800-1 is safe, has pharmacokinetic properties consistent with an ideal blood pool agent, and rapid elimination into urine after a single low-dose intravenous injection. Visualization of structure and function of the ureters starts within minutes after ZW800-1 injection and lasts several hours. Zwitterionic near-infrared fluorophores add value during laparoscopic abdominopelvic surgeries and could potentially decrease iatrogenic urethral injury. Moreover, ZW800-1 is engineered for one-step covalent conjugatability, creating possibilities for developing novel targeted ligands.

Safety, efficacy, and biomarker findings of PBT2 in targeting Abeta as a modifying therapy for Alzheimer's disease: a phase IIa, double-blind, randomised, placebo-controlled trial.

BACKGROUND: PBT2 is a metal-protein attenuating compound (MPAC) that affects the Cu2(+)-mediated and Zn2(+)-mediated toxic oligomerisation of Abeta seen in Alzheimer's disease (AD). Strong preclinical efficacy data and the completion of early, clinical safety studies have preceded this phase IIa study, the aim of which was to assess the effects of PBT2 on safety, efficacy, and biomarkers of AD. METHODS: Between December 6, 2006, and September 21, 2007, community-dwelling patients over age 55 years were recruited to this 12-week, double-blind, randomised trial of PBT2. Patients were randomly allocated to receive 50 mg PBT2, 250 mg PBT2, or placebo. Inclusion criteria were early AD (mini-mental state examination [MMSE] score between 20 and 26 points or Alzheimer's disease assessment scale-cognitive subscale (ADAS-cog) score between 10 and 25 points), taking a stable dose of acetylcholinesterase inhibitor (donepezil, galantamine, or rivastigmine) for at least 4 months, a modified Hachinski score of 4 points or less, and CT or MRI results that were consistent with AD. The principal outcomes were safety and tolerability. Secondary outcomes were plasma and CSF biomarkers and cognition. Analysis was intention to treat. The trial is registered with ClinicalTrials.gov, number NCT00471211. FINDINGS: 78 patients were randomly assigned (29 to placebo, 20 to PBT2 50 mg, and 29 to PBT2 250 mg) and 74 (95%) completed the study. 42 (54%) patients had at least one treatment emergent adverse event (10 [50%] on PBT2 50 mg, 18 [62%] on PBT2 250 mg, and 14 [48%] on placebo). No serious adverse events were reported by patients on PBT2. Patients treated with PBT2 250 mg had a dose-dependent (p=0.023) and significant reduction in CSF Abeta(42) concentration compared with those treated with placebo (difference in least squares mean change from baseline was -56.0 pg/mL, 95% CI -101.5 to -11.0; p=0.006). PBT2 had no effect on plasma biomarkers of AD or serum Zn(2+) and Cu(2+) concentrations. Cognition testing included ADAS-cog, MMSE, and a neuropsychological test battery (NTB). Of these tests, two executive function component tests of the NTB showed significant improvement over placebo in the PBT2 250 mg group: category fluency test (2.8 words, 0.1 to 5.4; p=0.041) and trail making part B (-48.0 s, -83.0 to -13.0; p=0.009). INTERPRETATION: The safety profile is favourable for the ongoing development of PBT2. The effect on putative biomarkers for AD in CSF but not in plasma is suggestive of a central effect of the drug on Abeta metabolism. Cognitive efficacy was restricted to two measures of executive function. Future trials that are larger and longer will establish if the effects of PBT2 on biomarkers and cognition that are reported here translate into clinical effectiveness.

Efficiency of assisted oocyte activation as a solution for failed intracytoplasmic sperm injection.

Failed fertilization after intracytoplasmic sperm injection (ICSI) can occur due to an oocyte activation defect. In these cases assisted oocyte activation (AOA) may help but efficiency is still unknown. Prior to AOA, the mouse oocyte activation test (MOAT) can be carried out by injecting human spermatozoa into mouse oocytes to evaluate their activating capacity. According to the MOAT activation percentage achieved, patients were classified into three groups: 0-20% (16 patients); 20-85% (seven patients); 85-100% (seven patients). For AOA, CaCl(2) was injected together with spermatozoa followed by a double Ca(2+) ionophore treatment. The fertilization rates before application of AOA in 50 cycles were 6%, 22% and 14% in, respectively, groups 1, 2 and 3 without any pregnancy. Fertilization and pregnancy rates after AOA in 61 cycles were significantly increased to 75% and 34% for group 1, 73% and 43% for group 2, and 75% and 17% for group 3 (P < 0.0001 and P < 0.004, respectively). Application of AOA results in normal fertilization and pregnancy rates in patients whose spermatozoa show deficient activation. When MOAT reveals no activation deficiency in spermatozoa, AOA still allows for high fertilization and acceptable pregnancy rates. The obstetric and neonatal outcomes after AOA were normal as no malformations were observed.

Production and health of cows given monensin prepartum and a high-energy diet postpartum.

The object of this study was to evaluate the impact of monensin administration on the early lactation performance of cows maintained on a high-energy diet, and on health traits during the transition period. Cows (n = 168; parity 3.3 +/- 1.4, initial body condition score 3.1 +/- 0.08, and milk yield of 34.3 kg/d +/- 0.9 for multiparous cows in the preceding lactation) were divided into control and monensin treatment groups. A controlled-release capsule supplying 335 mg of monensin/d for 95 d was inserted into the rumen of monensin-treated cows 30 d before the expected calving. Blood samples were obtained 2 h after feeding on d 14 prepartum and on d 7, 14, and 50 postpartum. Plasma glucose concentration was 3% higher (58 +/- 0.5 vs. 56.4 +/- 0.5 mg/dL) and beta-hydroxybutyric acid was 17% (6.7 +/- 0.3 vs. 8.0 +/- 0.3 mg/dL) lower in monensin-treated than control cows. Plasma glucose was 10% higher (60.0 +/- 0.6 vs. 54.5 +/- 0.3 mg/dL) and beta-hydroxybutyric acid was 16% lower (6.8 +/- 0.3 vs. 7.9 +/- 0.2 mg/dL) in primiparous than multiparous cows. Plasma nonesterified fatty acid concentration (measured only in primiparous cows) was 17% lower (287 +/- 15 vs. 336 +/- 17 muEq/L) in treated than in control cows. Rate of ketosis incidence was 60% lower (8 vs. 21%) in monensin-treated than in control cows, and the proportion of control cows that required a supply of glucogenic precursors was 3-fold higher than in monensin-treated cows. The body condition score was 3.1 +/- 0.05, 2.7 +/- 0.05, and 2.4 +/- 0.05 on d 60 prepartum and d 7 and 50 postpartum, respectively, and was not affected by treatment. During the first 5 mo of lactation, milk yield was 7% higher (37.6 +/- 0.6 vs. 35.2 +/- 0.6 kg/d) in monensin-treated cows than in control cows. Our results showed that monensin administration, as a controlled-release capsule in prepartum cows, can be beneficial, even if these cows are maintained on a high-energy diet during the subsequent lactation.

Effects on health, performance, and tissue residues of the ionophore antibiotic salinomycin in finishing broilers (21 to 38 d).

A study was conducted to evaluate the effects of feeding salinomycin at the recommended prophylactic level, and at 2 and 3 times this level, to finishing male broilers (d 21 to 38). Four treatment groups were given the experimental diets containing 0, 60, 120, or 180 parts per million (ppm) salinomycin from d 21 to 38. Performance, relative organ weights, selected serum enzymes, and salinomycin residues in liver, muscle, and serum were determined. Salinomycin supplementation had no effect on body weight, feed intake, or feed conversion, and caused no overt signs of toxicity. After a week of being fed the salinomycin diets, the serum activity of aspartate aminotransferase was significantly increased in chickens fed 180 ppm compared with controls. These birds also showed microscopic lesions in breast and thigh muscles, but not in cardiac muscle. Salinomycin residues were not detected by high-performance liquid chromatography coupled to tandem mass spectrometry in liver or muscle samples from the birds fed 0, 60, or 120 ppm salinomycin. However, chickens fed 180 ppm salinomycin had detectable levels in liver and muscle above the maximum residue level of 5 µg/kg established by the European Union. All birds fed salinomycin had salinomycin in their sera with levels ranging from N.D. (not detected) in the controls to 24.4 ± 7.9, 61.4 ± 18.9, and 94.5 ± 9.1 µg/L for salinomycin dietary levels of 60, 120, and 180 ppm, respectively. Serum salinomycin concentration was linearly related with salinomycin content in feed (y = 0.584x - 10, r2 = 0.999). The results showed that even at 3 times the prophylactic level, salinomycin does not induce clinical toxicosis or mortality. No salinomycin residues were found in edible tissues at the recommended dietary level or at 2 times this level. However, salinomycin was detected in serum regardless of the dietary level. A simple method for salinomycin determination in serum is described which can be used as a marker of exposure and/or to predict levels in the diet.

Manganese-52: applications in cell radiolabelling and liposomal nanomedicine PET imaging using oxine (8-hydroxyquinoline) as an ionophore.

The ionophore 8-hydroxyquinoline (oxine) has been used to radiolabel cells and liposomal medicines with 111In and, more recently, 89Zr, for medical nuclear imaging applications. Oxine has also shown promising ionophore activity for the positron-emitting radionuclide 52Mn that should allow imaging of labelled cells and nanomedicines for long periods of time (>14 days). However, to date, the radiometal complex formed and its full labelling capabilities have not been fully characterised. Here, we provide supporting evidence of the formation of [52Mn]Mn(oxinate)2 as the metastable complex responsible for its ionophore activity. The cell labelling properties of [52Mn]Mn(oxinate)2 were investigated with various cell lines. The liposomal nanomedicine, DOXIL® (Caelyx) was also labelled with [52Mn]Mn(oxinate)2 and imaged in vivo using PET imaging. [52Mn]Mn(oxinate)2 was able to label various cell lines with moderate efficiency (15-53%), however low cellular retention of 52Mn (21-25% after 24 h) was observed which was shown not to be due to cell death. PET imaging of [52Mn]Mn-DOXIL at 1 h and 24 h post-injection showed the expected pharmacokinetics and biodistribution of this stealth liposome, but at 72 h post-injection showed a profile matching that of free 52Mn, consistent with drug release. We conclude that oxine is an effective ionophore for 52Mn, but high cellular efflux of the isotope limits its use for prolonged cell tracking. [52Mn]Mn(oxinate)2 is effective for labelling and tracking DOXIL in vivo. The release of free radionuclide after liposome extravasation could provide a non-invasive method to monitor drug release in vivo.