Maintaining flippase activity in procoagulant platelets is a novel approach to reducing thrombin generation.

BACKGROUND: During thrombosis, procoagulant platelets expose phosphatidylserine (PS), which enhances local thrombin generation. Reducing platelet PS exposure could be a novel anti-thrombotic approach. PS is confined to the inner leaflet of the plasma membrane in unstimulated platelets by ATP-dependent "flippase" activity. Ca2+ ionophores trigger all platelets to expose a high level of PS by activating a scramblase protein and inactivating the flippase. Although R5421 was previously shown to reduce Ca2+ ionophore-induced PS exposure, its mechanism of action is unknown. OBJECTIVES: To determine the mechanism by which R5421 reduces platelet PS exposure. METHODS: Washed human platelets were stimulated with the Ca2+ ionophore, A23187, to induce procoagulant platelet formation while bypassing proximal receptor signalling. Platelets PS exposure was detected using annexin V or lactadherin in flow cytometry. NBD (7-nitro-2-1,3-benzoxadiazol-4-yl)-PS was used to assess scramblase and flippase activity. Thrombin generation was monitored using a fluorogenic substrate. RESULTS AND CONCLUSIONS: R5421 reduced the extent of A23187-stimulated platelet PS exposure, as demonstrated with annexin V or lactadherin binding. R5421 also maintained flippase activity in procoagulant platelets. Although R5421 appeared to inhibit scramblase activity in procoagulant platelets, it did not once the flippase had been inhibited, demonstrating that scramblase activity is not directly inhibited. Furthermore, R5421 inhibited the contribution of A23187-stimulated platelets to thrombin generation. Together these data demonstrate that R5421 reduces the extent of PS exposure in procoagulant platelets by maintaining flippase activity. Maintaining flippase activity in procoagulant platelets is a novel and effective approach to reducing thrombin generation.

Evidence of nigericin as a potential therapeutic candidate for cancers: A review.

Emerging studies have shown that nigericin, an H+, K+ and Pb2+ ionophore, has exhibited a promising anti-cancer activity in various cancers. However, its anti-cancer mechanisms have not been fully elucidated. In this review, the recent progresses on the use of nigericin in human cancers have been summarized. By exchanging H+ and K+ across cell membranes, nigericin shows promising anti-cancer activities in in vitro and in vivo as a single agent or in combination with other anti-cancer drugs through decreasing intracellular pH (pHi). The underlying mechanisms of nigericin also include the inactivation of Wnt/ß-catenin signals, blockade of Androgen Receptor (AR) signaling, and activation of Stress-Activated Protein Kinase/c-Jun N-terminal Kinase (SAPK/JNK) signaling pathways. In many cancers, nigericin is proved to specifically target putative Cancer Stem Cells (CSCs), and its synergistic effects on photodynamic therapy are also reported. Other mechanisms of nigericin including influencing the mitochondrial membrane potentials, inducing an increase in drug accumulation and autophagy, controlling insulin accumulation in nuclei, and increasing the cytotoxic activity of liposome-entrapped drugs, are also discussed. Notably, the potential adverse effects such as teratogenic effects, insulin resistance and eryptosis shall not be ignored. Taken together, these reports suggest that treatment of cancer cells with nigericin may offer a novel therapeutic strategy and future potential of translation to clinics.

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.

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.

Salinomycin and other ionophores as a new class of antimalarial drugs with transmission-blocking activity.

The drug target profile proposed by the Medicines for Malaria Venture for a malaria elimination/eradication policy focuses on molecules active on both asexual and sexual stages of Plasmodium, thus with both curative and transmission-blocking activities. The aim of the present work was to investigate whether the class of monovalent ionophores, which includes drugs used in veterinary medicine and that were recently proposed as human anticancer agents, meets these requirements. The activity of salinomycin, monensin, and nigericin on Plasmodium falciparum asexual and sexual erythrocytic stages and on the development of the Plasmodium berghei and P. falciparum mosquito stages is reported here. Gametocytogenesis of the P. falciparum strain 3D7 was induced in vitro, and gametocytes at stage II and III or stage IV and V of development were treated for different lengths of time with the ionophores and their viability measured with the parasite lactate dehydrogenase (pLDH) assay. The monovalent ionophores efficiently killed both asexual parasites and gametocytes with a nanomolar 50% inhibitory concentration (IC50). Salinomycin showed a fast speed of kill compared to that of standard drugs, and the potency was higher on stage IV and V than on stage II and III gametocytes. The ionophores inhibited ookinete development and subsequent oocyst formation in the mosquito midgut, confirming their transmission-blocking activity. Potential toxicity due to hemolysis was excluded, since only infected and not normal erythrocytes were damaged by ionophores. Our data strongly support the downstream exploration of monovalent ionophores for repositioning as new antimalarial and transmission-blocking leads.

Intoxicação espontânea de bovinos por Senna obtusifolia no Estado do Paraná / Spontaneous Senna obtusifolia poisoning in cattle in the state of Parana, Brazil

Plantas do gênero Senna causam miopatia degenerativa em bovinos e o maior número de relatos envolve Senna occidentalis. O objetivo desse trabalho é relatar, pela segunda vez no Brasil, um surto de intoxicação natural por Senna obtusifolia. É descrito um surto de intoxicação por Senna obtusifolia, na região noroeste do estado do Paraná, que aconteceu em um lote de 200 vacas, com idade entre 45 e 152 meses, introduzidas em um confinamento para melhorar a condição corporal antes do parto. A área do confinamento estava invadida pela planta e as vacas permaneceram no local durante oito dias. Entre quatro e nove dias após a entrada no confinamento 20 vacas adoeceram e somente uma se recuperou. Os sinais consistiram em mioglobinúria, incoordenação e decúbito esternal permanente. As vacas doentes apresentaram aumento das atividades das enzimas creatina quinase, aspartato aminotransferase, gamaglutamiltransferase e fosfatase alcalina. Lesões na musculatura esquelética dos membros posteriores caracterizadas por áreas pálidas representaram as principais alterações observadas à necropsia de quatro bovinos. Necrose segmentar multifocal da musculatura esquelética e necrose paracentral multifocal no fígado foram as alterações histopatológicas mais relevantes. As evidências epidemiológicas, clínicas e patológicas indicam o diagnóstico de intoxicação por Senna obtusifolia. A planta demonstrou ter efeito miotóxico e hepatotóxico nos animais intoxicados e a doença foi quase sempre fatal.

Plants of the genus Senna cause a degenerative myopathy in cattle and most of the reports refer to Senna occidentalis. The aim of this paper is to report, for the second time in Brazil, an outbreak of natural poisoning by Senna obtusifolia. It happened in the northwestern Paraná in a herd of 200 cows, 45 to 152 months of age, which had been placed into a feedlot to improve their nutritional status before the calving period. The cows stayed for eight days in this feedlot infested by the plant. Four to nine days after they got into the feedlot, 20 cows became ill and only one recovered. The clinical signs consisted of myoglobinuria, incoordination and permanent sternal recumbency. The affected cows showed increased activity of creatine phosphokinase, aspartate aminotransferase, gama-glutamyltransferase, and alkaline phosphatase. The main postmortem changes were in skeletal muscles of the hind limbs characterized by pale areas. The histological alterations were multifocal segmental necrosis of skeletal muscles and hepatic multifocal paracentral necrosis. Epidemiological, clinical and pathological data led to the diagnosis of Senna obtusifolia poisoning. The plant showed miotoxic and hepatotoxic effects on the poisoned animals and the disease was almost always lethal.

A phase I study of the metal ionophore clioquinol in patients with advanced hematologic malignancies.

UNLABELLED: Clioquinol is a small-molecule metal ionophore that inhibits the proteasome through a metal-dependent mechanism. Here, we report a phase I study of clioquinol in patients with refractory hematologic malignancies. Neuropathy and abdominal pain were dose-limiting toxicities. Minimal pharmacodynamic effects were observed, and there were no clinical responses. BACKGROUND: Clioquinol is a small-molecule metal ionophore that inhibits the enzymatic activity of the proteasome and displays preclinical efficacy in hematologic malignancies in vitro and in vivo. Therefore, we conducted a phase I clinical trial of clioquinol in patients with refractory hematologic malignancies to assess its safety and determine its biological activity in this patient population. METHODS: Patients with refractory hematologic malignancies were treated with increasing doses of oral clioquinol twice daily for 15 doses. Plasma and intracellular levels of clioquinol were measured. Enzymatic activity of the proteasome was measured before and after drug administration. RESULTS: Sixteen cycles of clioquinol were administered to 11 patients with 5 patients reenrolled at the next dose level as per the permitted intrapatient dose escalation. Dose-limiting neurotoxicity and abdominal pain were observed at a dose of 1600 mg twice daily. Intracellular drug levels were low. Minimal inhibition of the proteasome was observed. No clinical responses were observed. CONCLUSION: In patients with refractory hematologic malignancies, the maximal tolerated dose of clioquinol was determined. Minimal inhibition of the proteasome was observed at tolerable doses, likely due to low intracellular levels of the drug.

Surtos de intoxicação por salinomicina em chinchilas (Chinchilla lanigera) / Outbreaks of salinomycin toxicosis in Chinchillas (Chinchilla lanigera)

Quatro surtos de intoxicação por salinomicina são descrito em chinchilas de três municípios do Estado do Rio Grande do Sul. Uma semana após a ingestão de ração contendo 37 ppm de salinomicina, aproximadamente duas mil chinchilas de quatro fazendas expostas diminuíram o consumo da ração. Quatrocentos e vinte sete chinchilas demonstraram apatia. Dessas, duzentos e setenta e sete desenvolveram decúbito esternal e lateral, dispneia e coma, seguidos de morte. As primeiras mortes ocorreram oito dias após a ingestão da ração. A evolução dos sinais clínicos até a morte ou eutanásia foi de 2-5 dias. Os exames bioquímicos do soro sanguíneo em quatro chinchilas revelaram níveis aumentados da alanina aminotransferase, aspartato transaminase, fosfatase alcalina, creatina cinase, glicose, triglicerídeos e colesterol total. Quarenta e cinco chinchilas foram submetidas à necropsia. Os achados macroscópicos consistiam de marcada lipidose hepática em todas as chinchilas necropsiadas; fetos em estado de decomposição em doze chinchilas que estavam prenhes. Microscopicamente, múltiplas fibras musculares esqueléticas estavam hipereosinofílicas, tumefeitas e com perda das estriações. Nas chinchilas que sobreviveram por mais dias era possível observar segmentos fragmentados de miofibras afetadas (necrose flocular) e regeneração de miofibras. No fígado foi observada marcada degeneração gordurosa. Não foram observadas anormalidades microscópicas nos demais órgãos analisados. Análises à procura de aflatoxinas, resíduos de pesticidas e isolamento bacteriano foram negativos. A análise da ração por cromatografia líquida revelou 37ppm de salinomicina na ração. A ração suspeita foi administrada a 12 chinchilas, três das quais (25 por cento) morreram apresentando lesões semelhantes às observadas nas chinchilas com a doença natural. O diagnóstico de intoxicação por salinomicina foi baseado na epidemiologia, lesões histológicas características e na presença de salinomicina na ração administrada nas quatro criações envolvidas.

Four outbreaks of ionophore toxicosis are described in chinchillas from four commercial farms located in three municipalities in the state of Rio Grande do Sul, southern Brazil. Approximately 2,000 chinchillas showed decrease in food intake one week after start ingesting a ration containing 37 ppm of salinomycin. Four hundred and twenty seven chinchillas showed apathy. Of those 277 develop sternal and lateral recumbence, dyspnea and coma followed by death. First deaths occurred eight days after the start on the salinomycin containing ration; clinical course was 2-5 days. Serum chemistry carried out in four chinchillas revealed increased levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, creatinenin kinase, glucose, triglicerids and total cholesterol. Forty five affected chinchillas were necropsied; consistent necropsy findings were marked hepatic lipidosis; additionally twelve pregnant chinchillas had dead decomposing fetuses. Microscopically skeletal muscles had multifocally swollen hypereosinophilic myofibers with loss of cross striations. In those chinchillas that survived longer than a few days, microscopic features in the skeletal muscle included segmental fragmentation of dead fibers (floccular necrosis) and myofiber regeneration. Marked fatty degeneration was observed in the livers of all affected chinchillas. No microscopic changes were observed in other organs. Chemical analysis in the feed consumed by the chinchillas did not detect aflatoxins or pesticides residues; bacterial culture performed in samples of liver and intestinal contents from necropsied chinchillas yielded no significant bacterial growth. Analysis by thin layer chromatography performed in the ration consumed by the chinchillas detected 37 ppm of salinomycin. The suspected ration was fed to 12 chinchillas three of which (25 percent) died with similar lesions to those observed in the natural cases. The diagnosis of salinomycin toxicosis was based in the epidemiology, histology of the lesions, on the detection of significant amounts of salinomycin in the ration used to feed the chinchillas in the four involved farms and on the reproduction of disease by feeding the suspected ration to susceptible chinchillas.

[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).

Intoxicação por antibióticos ionóforos em animais:[revisão] / Ionophore poisoning in animals: [review]

O uso terapêutico de antibióticos ionóforos em medicina veterinária difundiu-se muito nos últimos anos, com conseqüente aumento no risco de intoxicação em animais. Antibióticos ionóforos são usados como coccidiostáticos e como aditivo em alimentos para animais, com o propósito de estimular o desenvolvimento e o ganho de peso. Os ionóforos mais utilizados na alimentação de animais são a monensina, lasalocida, nasarina e salinomicina. Há uma grande variação na susceptibilidade dos efeitos tóxicos dos ionóforos de acordo com a espécie animal. A intoxicação pode ocorrer quando dosagens elevadas de ionóforos são adicionadas aos alimentos, ou quando ionóforos são incluídos inadvertidamente ou acidentalmente em dosagens não corretas para determinada espécie animal. Casos de intoxicação têm sido descritos em bovinos, ovinos, suínos, eqüinos, cães e aves. Para os eqüinos os ionóforos são extremamente tóxicos. São considerados seguros quando usados nas espécies-alvo, dentro das dosagens recomendadas pelo fabricante.

The therapeutic use of ionophores in veterinary medicine has grown in the last years, with resultant increase in the risk of poisoning in animals. Ionophores are used as food additives as coccidiostacts in several animal species and growth promoter and bloat prevention in ruminants. The most often used ionophores are monensin, lasalocid, narasin and salinomycin. There is a great variation in the susceptibility to the toxic effect of ionophores in different animal species. Poisoning can occur when the dosage is too high or when not correct doses for a certain animal species are given. Cases of poisoning have been described in sheep, swine, horses, dogs and poultry. For horses ionophores are extremely toxic. The use of ionophores is only safe when used accordingly to the instructions of the manufacturer and especially for each animal species. In this paper the most important data regarding clinical-pathological and pathogenic aspects, and also the conditions in which the poisoning may occur are critically reviewed.

Parâmetros digestivos e produção de proteína microbiana em novilhas em crescimento compensatório / Digestive parameters and protein microbial production in heifers under compensatory growth

Avaliaram-se o manejo para crescimento compensatório e o efeito da suplementação com ionóforo na dieta sobre os parâmetros digestivos e sobre a produção de proteína microbiana de novilhas leiteiras. Foram utilizadas 20 animais puros da raça Pardo-Suíça, com média de peso inicial de 200kg aos cinco meses de idade. Os tratamentos foram arranjados em um esquema fatorial 2x2x2, e os animais foram alocados, aleatoriamente, em cada uma das combinações. O fator 1 consistiu dos sistemas de alimentação (convencional e crescimento compensatório), o fator 2, da utilização (200mg de monensina/animal/dia) ou não de ionóforo e o fator 3, dos períodos de alimentação (P1 e P2). A inclusão de ionóforo na dieta aumentou os coeficientes de digestibilidade total da matéria seca, da matéria orgânica, dos carboidratos totais e da fibra em detergente neutro. Não houve efeito do sistema de alimentação, da adição de ionóforo à dieta e do período sobre a produção microbiana. A eficiência microbiana (g PB microbiana/kg de NDT consumido) no período de restrição foi maior que no período de realimentação.

The effects of compensatory growth and ionophore supplementation of diet of dairy heifer on digestive parameters and protein microbial production were evaluated. Twenty five-month-old Brown-Swiss heifers averaging 200kg b.w. were used. The treatments were arranged in a factorial design (2x2x2) with the animals randomly allocated to each of the combinations. Factor 1 was based on the feeding systems (conventional and compensatory growth), factor 2 on ionophore supplementation option (200mg of monensin/animal/day or not) and factor 3, on the feeding periods (P1 and P2). The diet supplemented with ionophore increased the total digestibility coefficients of dry matter, organic matter, total carbohydrates, and neutral detergent fiber. No effect of feeding systems, ionophore supplementation, or feeding periods based on microbial production was oberved. The microbial efficiency (g of microbial crude protein/kg of NDT intake) during the restriction period was higher than the re-feeding period.

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.

Efeitos da adição de propilenoglicol ou monensina à silagem de milho sobre a cinética de degradação dos carboidratos e produção cumulativa de gases in vitro / Effects of adding propylene glycol or monensin to corn silage on the degradation kinetics of carbohydrates and in vitro cumulative gas production

Avaliaram-se os efeitos dos aditivos propilenoglicol e/ou monensina sobre a degradabilidade média e efetiva dos carboidratos totais, pH e produção cumulativa de gases da silagem de milho por meio da técnica in vitro semi-automática de produção de gases. Os tratamentos constituíram-se de silagem de milho (SM); SM associada ao propilenoglicol (SM+PG); SM associada à monensina (SM+MO); SM associada ao propilenoglicol e à monensina (SM+PG+MO) avaliados com duas, quatro, seis, 12, 24, 48 e 96 horas. A adição de monensina ou monensina associada ao propilenoglicol aumentou (P<0,05) a degradabilidade dos carboidratos totais às duas horas. SM+MO apresentou maior degradabilidade efetiva dos carboidratos totais em todas as taxas de passagem. A utilização de monensina reduziu a produção cumulativa de gases das 12 às 96 horas. Entre os tratamentos, SM+MO apresentou o menor potencial de produção de gases (221ml/g carboidratos totais) e o menor tempo de colonização (1,08 horas) em relação aos tratamentos SM e SM+PG (1,58 e 1,49 horas, respectivamente). A produção cumulativa de gases e degradabilidade dos carboidratos totais apresentaram elevada correlação, variando de 94 a 97 por cento (P<0,01). O pH do meio foi inversamente correlacionado à degradabilidade dos carboidratos totais (r= -79 por cento, P<0,01). O uso de monensina pode ser uma boa alternativa para se melhorarem os parâmetros da cinética de degradação da silagem de milho.

The effects of the additives propylene glycol and/or monensin on the degradation of total carbohydrates, pH, and cumulative gas production of corn silage by the semi-automated in vitro gas production technique were evaluated. The treatments were corn silage (CS); CS plus propylene glycol (CS+PG); CS plus monensin (CS+MO), and CS plus propylene glycol and monensin (CS+PG+MO), which were evaluated at two, four, six, 12, 24, 48, and 96 hours. The addition of monensin or monensin plus propylene glycol increased (P<0.05) the degradation of total carbohydrates at 2h. The effective degradations of total carbohydrates for CS+MO treatment (55.2; 42.7; and 36.5 percent) were the highest in all passage rates. The use of monensin reduced cumulative gas production from 12 to 96h. CS+MO treatment had the lowest potential of gas production (221ml/g total carbohydrates), and the lowest Lag phase (1.08h), as compared to CS and CS+PG treatment (1.58 and 1.49h, respectively). Cumulative gas production and degradation of total carbohydrates were highly correlated (94 to 97 percent; P<0.01). The pH was inversely correlated to degradability of total carbohydrates (r= -0.79; P<0.01). Thus, monensin may be used for improving the ruminal degradability of corn silage.

A meta-analysis of the impact of monensin in lactating dairy cattle. Part 3. Health and reproduction.

A meta-analysis of the impact of monensin on health and reproductive outcomes in dairy cattle was conducted. A total of 16 papers were identified with sufficient data and quality to evaluate health and reproductive outcomes for monensin. The available trials provided approximately 9,500 cows with sufficient data for analysis. This provided good statistical power to examine the effects of monensin on health and reproduction. Over all the trials analyzed, monensin decreased the risk of ketosis [relative risk (RR) = 0.75], displaced abomasums (RR = 0.75), and mastitis (RR = 0.91). No significant effects of monensin were found for milk fever, lameness, dystocia, retained placenta, or metritis. Monensin had no effect on first-service conception risk (RR = 0.97) or days to pregnancy (hazard ratio = 0.93). However, the impact of monensin on dystocia, retained placenta, and metritis was heterogeneous for all 3 outcome measures and random effect models were utilized. Causes of the heterogeneity were explored with meta-regression. Days of treatment with monensin before calving increased the risk of dystocia. Delivery method of monensin influenced the incidence of retained placenta and metritis, with risk being lower with controlled release capsule treatment compared with delivery in either topdress or in a total mixed ration. Days of treatment before calving also influenced retained placenta with an increase in risk with more days treated before calving. Improvements in ketosis, displaced abomasums, and mastitis with monensin were achieved. Exposure to prolonged treatment in the dry period with monensin may increase the risk of dystocia and retained placenta.

Oxidative effects of Na+--specific ionophore monensin on the rat epididymis.

The carboxylic antibiotic ionophore monensin is well-known for the Na+/H+ exchanger activity across the biological membranes. The current study has been designed to investigate the effect of monensin on spermatozoal concentration, motility, and oxidative stress-related parameters in the rat epididymis. Monensin was administered orally at a dose of 3.5 mg/kg body weight daily for 70 days, a duration that coincides with the completion of the spermatogenic cycle. At the end of the respective treatment, the epididymis was isolated into three separate regions--the capitum, corpus, and the cauda--successively away from the head of the testis. Marked changes were noted in the body weight, organ (epididymis) weight, sperm concentration and motility, as well as the morphologic observations of the sperm and the histologic architecture of the epididymal epithelium. Significant alterations were also recorded in the oxidative stress parameters such as the lipid peroxidation product, malonyldialdehyde, and the activity of superoxide dismutase, glutathione sulfotransferase, glutathione reductase, and catalase. The nonenzymatic thiol content such as the total, oxidized, and reduced glutathione showed significant changes and the tissue phosphatases such as alkaline and acid phosphatase were increased, indicative of the interference of the drug in lysosomal and Golgi membrane complex. The findings of the current study indicate interactions during the spermatozoal maturational process in the epididymis, and a significant potential use of monensin in male contraception may be suggested.

Intoxicação experimental por monensina em búfalos e bovinos / Experimental monensin poisoning in water buffaloes (Bubalus bubalis) and cattle

Sinais clínicos e lesões característicos de intoxicação por monensina foram induzidos em búfalos dosados (1 dia) com 15, 10, 7,5 e 5mg/kg de monensina. Apenas os búfalos dosados com 2,5 (1 dia) e 1 mg/kg (7 dias) de monensina não morreram. Os sinais clínicos iniciaram cerca de 6 h após dosagem com monensina e incluíram apatia, anorexia, diarréia, sialorréia, fraqueza muscular, taquicardia, dificuldade locomotora, dispnéia, distensão da jugular, decúbito e morte. As dosagens de creatinina quinase (CK) dos búfalos aumentaram acentuadamente após dosagem com monensina. As alterações macroscópicas foram ascite, hidrotórax, hidropericárdio, cardiomegalia, hepatomegalia e áreas pálidas focais no miocárdio e nos músculos esqueléticos. Degeneração e necrose de miofibras foram os principais achados histopatológicos. Por outro lado, nenhuma evidência de doença, nem mesmo alteração nos níveis de CK, foram observados nos bovinos dosados com as mesmas dosagens de monensina, confirmando observações preliminares que esses animais são mais resistentes à monensina que os búfalos.

Monensin is widely used as a feed additive to improve performance of livestock; however accidental poisoning by this ionophore compound has been reported in a number of animal species. Typical clinical signs and lesions of monensin poisoning were induced in water buffaloes dosed with single dosages of 15, 10, 7.5, and 5mg/kg of the compound. Only buffaloes dosed with 2.5 mg/kg (1 day) and 1mg/kg (7 days) survived. Clinical signs initiated about 6 h post-dosing and included apathy, anorexia, diarrhea, drooling, muscular weakness, locomotion disorders, dyspnea, tachycardia, jugular distension and pulse, recumbency and death. The creatine kinase (CK) levels were highly augmented in blood samples of buffaloes dosed with monensin. Most prominent gross changes were ascites, hydrothorax, hydropericardium, cardiomegaly, hepatomegaly, and focal pale areas in the myocardium and in skeletal muscles. Degeneration and necrosis of myofibers were the principal histopathological findings. Conversely, no evidence of disease, neither change in CK levels were observed in the beef cattle steers dosed with same doses, confirming preliminary findings that buffaloes are more susceptible to monensin than cattle. In addition, this communication presents the minimal toxic dosage of monensin to buffaloes and suggests that CK tests may serve as health monitoring tools in the management of buffalo herds supplemented with monensin.

Ionophores: their use as ruminant growth promotants and impact on food safety.

Ionophores (such as monensin, lasalocid, laidlomycin, salinomycin and narasin) are antimicrobial compounds that are commonly fed to ruminant animals to improve feed efficiency. These antimicrobials specifically target the ruminal bacterial population and alter the microbial ecology of the intestinal microbial consortium, resulting in increased carbon and nitrogen retention by the animal, increasing production efficiency. Ionophores transport ions across cell membranes of susceptible bacteria, dissipating ion gradients and uncoupling energy expenditures from growth, killing these bacteria. Not all bacteria are susceptible to ionophores, and several species have been shown to develop several mechanisms of ionophore resistance. The prophylactic use of antimicrobials as growth promotants in food animals has fallen under greater scrutiny due to fears of the spread of antibiotic resistance. Because of the complexity and high degree of specificity of ionophore resistance, it appears that ionophores do not contribute to the development of antibiotic resistance to important human drugs. Therefore it appears that ionophores will continue to play a significant role in improving the efficiency of animal production in the future.

Short communication: Effects of a monensin premix on milk fatty acid content during subacute ruminal acidosis in dairy cows.

The effects of a monensin premix on milk fatty acid content during grain-induced subacute ruminal acidosis (SARA) in Holstein cows receiving a total mixed ration was investigated. Six multiparous, rumen-fistulated Holstein cows were used in a two-treatment, two-period crossover design with 6-wk periods. Experimental treatments were either a monensin premix or a placebo premix. At the beginning of wk 4, SARA was induced in experimental cows for a 10-d period using a grain challenge model. The administration of a monensin premix elevated milk fat proportion of total short-chain saturated fatty acids (sum of C4 to C15). Milk fat proportions of conjugated linoleic acid isomers were unaffected. Linolenic acid (C18:3n3) proportion in milk fat of monensin-treated cows were lower when compared with placebo-treated cows during the SARA period. Results from this study indicate that dietary supplementation with monensin during SARA had little effect on milk fatty acid content.