Comparative and Functional Analyses Reveal Conserved and Variable Regulatory Systems That Control Lasalocid Biosynthesis in Different Streptomyces Species.

Lasalocid, a representative polyether ionophore, has been successfully applied in veterinary medicine and animal husbandry and also displays promising potential for cancer therapy. Nevertheless, the regulatory system governing lasalocid biosynthesis remains obscure. Here, we identified two conserved (lodR2 and lodR3) and one variable (lodR1, found only in Streptomyces sp. strain FXJ1.172) putative regulatory genes through a comparison of the lasalocid biosynthetic gene cluster (lod) from Streptomyces sp. FXJ1.172 with those (las and lsd) from Streptomyces lasalocidi. Gene disruption experiments demonstrated that both lodR1 and lodR3 positively regulate lasalocid biosynthesis in Streptomyces sp. FXJ1.172, while lodR2 plays a negative regulatory role. To unravel the regulatory mechanism, transcriptional analysis and electrophoretic mobility shift assays (EMSAs) along with footprinting experiments were performed. The results revealed that LodR1 and LodR2 could bind to the intergenic regions of lodR1-lodAB and lodR2-lodED, respectively, thereby repressing the transcription of the lodAB and lodED operons, respectively. The repression of lodAB-lodC by LodR1 likely boosts lasalocid biosynthesis. Furthermore, LodR2 and LodE constitute a repressor-activator system that senses changes in intracellular lasalocid concentrations and coordinates its biosynthesis. LodR3 could directly activate the transcription of key structural genes. Comparative and parallel functional analyses of the homologous genes in S. lasalocidi ATCC 31180T confirmed the conserved roles of lodR2, lodE, and lodR3 in controlling lasalocid biosynthesis. Intriguingly, the variable gene locus lodR1-lodC from Streptomyces sp. FXJ1.172 seems functionally conserved when introduced into S. lasalocidi ATCC 31180T. Overall, our findings demonstrate that lasalocid biosynthesis is tightly controlled by both conserved and variable regulators, providing valuable guidance for further improving lasalocid production. IMPORTANCE Compared to its elaborated biosynthetic pathway, the regulation of lasalocid biosynthesis remains obscure. Here, we characterize the roles of regulatory genes in lasalocid biosynthetic gene clusters of two distinct Streptomyces species and identify a conserved repressor-activator system, LodR2-LodE, which could sense changes in the concentration of lasalocid and coordinate its biosynthesis with self-resistance. Furthermore, in parallel, we verify that the regulatory system identified in a new Streptomyces isolate is valid in the industrial lasalocid producer and thus applicable for the construction of high-yield strains. These findings deepen our understanding of regulatory mechanisms involved in the production of polyether ionophores and provide novel clues for the rational design of industrial strains for scaled-up production.

Streptomyces lasalocidi sp. nov. (formerly 'Streptomyces lasaliensis'), an actinomycete isolated from soil which produces the polyether antibiotic lasalocid.

Strain ATCC 31180T was isolated from soil collected in Hyde Park, Massachusetts (USA), and found to produce the polyether antibiotic lasalocid. The name 'Streptomyces lasaliensis' has been in common use since 1974, without a recognized taxonomic description. The most closely related type cultures determined by rRNA gene sequence similarity were Streptomyces longwoodensis DSM 41677T (100 %) and Streptomyces galbus DSM 40089T (100 %). OrthoANI values with S. longwoodensis and S. galbus were 95.50 and 94.41 %, respectively. Chemotaxonomic characteristics supported inclusion within the genus Streptomyces. The cell wall peptidoglycan contained ll-diaminopimelic acid, and the major whole-cell sugars were glucose and ribose. Polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol, one unidentified lipid and one unidentified glycolipid. The major menaquinones detected were MK9(H4), MK9(H6) and MK9(H8). The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0, iso-C15 : 0 and anteiso-C17 : 1. Its DNA had a G+C content of 72.6 %. Differentiation of ATCC 31180T from the closely related species was evident from digital DNA-DNA hybridization values of 61.80 and 56.90 % for S. longwoodensis and S. galbus respectively. Significant differences were seen in the polyphasic phenotypic analyses. ATCC 31180T produced lasalocid, grew from 10 to 45 °C, pH4-8 and in the presence of 0-10 % NaCl, 0.01 % NaN3 and 1 % phenol. Melanin was produced; H2S and indole were not. Nitrate was not reduced. Spore chains were retinaculum-apertum and spore surfaces were smooth. Spore colour, mycelia colour and soluble pigment production were medium-dependent. The proposed name is Streptomyces lasalocidi sp. nov.; the type strain being ATCC 31180T (=NRRL 3382T=DSM 46487T).

Trypanosoma brucei: trypanocidal and cell swelling activities of lasalocid acid.

Chemotherapeutic treatment of human and animal trypanosomiasis is unsatisfactory because only a few drugs are available. As these drugs have poor efficacy and cause adverse reactions, more effective and tolerable medications are needed. As the polyether ionophore antibiotic lasalocid acid is used as medicated feed additive in cattle, the compound was tested for its trypanocidal and cytotoxic activity against bloodstream forms of Trypanosoma brucei and human myeloid HL-60 cells. The concentrations required of lasalocid acid to reduce the growth rate of trypanosomes by 50% and to kill the parasites were 1.75 and 10 µM, respectively. The ionophore displayed also cytotoxic activity against HL-60 cells but the human cells were about 10 to 14 times less sensitive indicating moderate selectivity. As the trypanocidal mechanism of action of polyether ionophore antibiotics is due to a sodium influx-induced cell swelling, the effect of lasalocid acid on cell volume change in bloodstream-form trypanosomes was investigated. Interestingly, lasalocid acid induced a much faster cell swelling in trypanosomes than the more trypanocidal related ionophore salinomycin. These results support further investigations of lasalocid acid and derivatives thereof as potential agents against African trypanosomiasis.

Synergistic inhibition of Aß production by combinations of γ-secretase modulators.

Alzheimer's disease is associated with the accumulation of amyloid-ß (Aß) in the brain. In particular, the 42-amino acid form, Aß1-42, is thought to play a key role in the disease. It is therefore of interest that diverse compounds, known as γ-secretase modulators (GSM), can selectively decrease Aß1-42 production without inhibiting the production of other forms of Aß. Here we describe the novel discovery of synergistic inhibition of Aß by certain combinations of GSMs. Cell cultures were treated with pairwise combinations of GSMs to determine how Aß peptide production was affected. Analysis of isobolograms and calculation of the combination index showed that BMS-869780 and GSM-2 were highly synergistic. Additional combinations of GSMs revealed that inhibition of Aß occurred only when one GSM was of the "acid GSM" structural class and the other was of the "non-acid GSM" class. A total of 15 representative acid/non-acid GSM combinations were shown to inhibit Aß production, whereas 10 pairwise combinations containing two acid GSMs or containing two non-acid GSMs did not inhibit Aß. We also discovered that lasalocid, a natural product, is a potent GSM. Lasalocid is unique in that it did not synergize with other GSMs. Synergism did not translate in vivo perhaps because of biochemical differences between the cell culture model and brain. These findings reinforce the pharmacological differences between different structural classes of GSMs, and may help to exploit the potential of γ-secretase as a drug target.

Lasalocid induces cytotoxic apoptosis and cytoprotective autophagy through reactive oxygen species in human prostate cancer PC-3 cells.

Lasalocid is an antibiotic from the group of carboxylic ionophores, produced by Streptomyces lasaliensis. But there was limited information of lasalocid on human prostate cancer cells. In the present studies, to better understand its effect in human prostate cancer cells, apoptosis and autophagy associated with possible signal pathways in vitro was examined. Our study showed that lasalocid mediated cell cycle arrest in G0/G1 phase by reducing G1 phase dependent proteins, indicating entering into apoptotic cell death pathway. Lasalocid-induced apoptosis was involved with reactive oxygen species (ROS) production, and mitochondrial hyperpolarization. In addition, lasalocid induced autophagy through microtubule-associated protein 1 light chain 3 (LC-3)-II conversion, acidic vesicular organelles formation and GFP-LC-3 punctuate, which was inhibited by 3-methyladenine (3-MA), a widely used pharmacological inhibitor of autophagy. Furthermore, the autophagic phenomena were mediated by production of ROS, confirming that inhibition of ROS with N-acetyl-l-cysteine, a ROS inhibitor, attenuated lasalocid-triggered autophagy. Inhibition of autophagy with 3-MA enhanced the lasalocid-induced apoptosis through enhanced ROS generation. Taken together, lasalocid should be useful in the search for new potential chemotherapeutic agents for understanding the molecular mechanisms of anticancer in prostate cancer cells.

Avaliação da eficácia da lasalocida e de alguns fatores epidemiológicos de Eimeria spp. parasitando bezerros Nelore mantidos em regime de pastejo / Effectiveness of lasalocid assessment and some epidemiological factors of Eimeria spp. parasitizing Nellore calves kept on pasture

A principal importância da eimeriose em bovinos, se deve ao baixo desempenho produtivo que os animais demonstram quando esta enfermidade apresenta-se sob a forma sub-clínica. Como objetivos, o presente trabalho avaliou a eficácia do uso da lasalocida sódica contra espécies de Eimeria spp. parasitando bezerros; avaliou também o desempenho ponderal dos animais submetidos aos diferentes tratamentos e analisou alguns fatores epidemiológicos que possam interferir na infecção por Eimeria nos bezerros. Foram utilizados 288 bezerros no dia 0 do estudo. Os animais pertencentes ao tratamento 01 receberam sal mineral proteinado de baixo consumo sem adição de lasalocida, enquanto que os bezerros do Tratamento 02 sal mineral proteinado de baixo consumo, com adição de lasalocida sódica, administrado via oral para bezerros dos quatro/cinco/seis meses até dez meses de idade. Colheita de fezes e pesagem dos animais foram realizadas nos dias 0 (antes do início do experimento), na desmama, 30 e 60 dias após desmama (DPD). A avaliação de alguns fatores epidemiológicos que pudessem ser relacionados com a infecção por Eimeria spp nos bezerros, como o desmame, sexo e época do ano, foram analisados neste estudo, levando-se em consideração os resultados encontrados durante todo estudo, para os 144 animais pertencentes ao grupo controle. Foram identificadas nove espécies de Eimeria nos bezerros em ordem decrescente: E. brasiliensis, E. wyomingensis, E. bovis, E. canadenses, E. zuernii, E. auburnensis, E. ellipsoidalis, E. pellita e E. cylindrica. Inesperadamente, diminuição na carga parasitária dos animais pode ser observada após o desmame. Mesmo a fazenda não adotando medidas de manejo que visam maior produtividade como a Inseminação Artificial em Tempo Fixo, que por sua vez acaba aumentando o número de nascimentos e unidade animal/hectare em uma determinada época do ano, elevado parasitismo pelo coccídio em questão foi diagnosticado nos bezerros pertencentes ao grupo controle. Talvez a época do ano em que o estudo foi realizado pode ter influenciado neste aspecto. As contagens de oocistos por grama (OoPG) de fezes para Eimeria dos animais tratados com lasalocida foram estatisticamente inferiores (P≤ 0,05) as do grupo controle após o início do estudo. O composto alcançou eficácia ≥ 95% contra o parasito em questão. No final do estudo, os animais que receberam lasalocida ganharam em média, 7,2kg a mais (P≤ 0,05) que os bezerros pertencentes ao grupo controle. Em propriedades que tem como objetivo a venda de bezerros logo após a desmama, recomenda-se o início do tratamento com a lasalocida, junto ao creep-feeding, a partir de três messes de idade, uma vez que diferencial no ganho em peso médio dos bezerros tratados foi significativamente (P≤ 0,05) mais elevado, em comparação ao grupo controle, após cinco meses de tratamento com o referido composto. Apesar de a lasalocida ser utilizada como um aditivo alimentar para animais, a diferença no ganho em peso vivo médio entre animais tratados com a lasalocida, em comparação a animais pertencentes ao grupo controle, também pode ser relacionada, em partes, a infecção dos animais por Eimeria spp., conforme discutido neste artigo, entretanto, futuros estudos devem ser conduzidos para comprovar esta hipótese.(AU)

The main importance of eimeriosis in cattle is due to lower performance shown with the disease in its sub-clinical form. This study evaluated the efficacy of lasalocid used against Eimeria spp. parasitizing calves. We also evaluated the weight gain of calves submitted to different treatments and analyzed some epidemiological factors that might interfere with Eimeria infection; 288 calves were used in the study. The calves of treatment 1 received protein mineral salt in low consumption without lasalocid, while the calves of treatment 2 received protein mineral salt on low consumption with lasalocid, administered orally to 4 to 10-month-old calves. Harvest of feces and weight control was made on days 0 (before the start of the experiment), at weaning, and 30 and 60 days after weaning (DAW). Evaluation of some epidemiological factors which could be related to infection by Eimeria spp. of the calves, such as weaning, sex and time of year, were analyzed, taking into account the results regarding the 144 calves of the control group. Nine species of Eimeria were identified in descending order: E. brasiliensis, E. wyomingensis, E. bovis, E. canadian, E. zuernii, E. auburnensis, E. ellipsoidalis, E. pellita and E. cylindrica. Unexpectedly, decrease in parasite load could be observed after weaning. Even the farm did not adopt management measures aimed for greater productivity, as Artificial Insemination in Fixed Time, which in turn ends up with increase of the number of births and animal unit per hectare at a certain period of year, high parasitism of coccidia was diagnosed in calves of the control group. Oocyst counts per gram (OPG) of calves treated with lasalocid were significantly lower (P≤ 0.05) in the control group. The compound achieved ≥ 95% efficacy against the parasite in question. At the end of the study, calves fed lasalocid gained on average 7.2kg (p≤ 0.05) more than calves in the control group. For a farm that aims to sell calves soon after weaning, is recommended to start treatment with lasalocid, with the creep-feeding, from an age of three months on, since the weight gain calves treated with lasalocid was significantly (p=0.05) higher compared with the weight gain of the control group after five months of treatment. The difference in weight gain of calves treated with lasalocid compared with caves in the control group may also be partially related to the infection by Eimeria spp., as discussed in this paper.(AU)

Safety and efficacy of Avatec® 150G (lasalocid A sodium) for chickens for fattening and chickens reared for laying, and modification of the terms of authorisation for chickens for fattening, chickens reared for laying, turkeys for fattening, minor avian species (pheasants, guinea fowl, quails and partridges) except laying birds.

Avatec® 150G is an anticoccidial feed additive containing 15% of the active ingredient lasalocid A sodium, an ionophore anticoccidial. The tolerance study indicated that oral administration of lasalocid A sodium via feed at doses at and above the targeted lasalocid dose of 125 mg/kg was not tolerated in chickens for fattening. Consequently, no safe dose can be established by the tolerance study submitted. Concurrent administration of lasalocid with tiamulin and certain other medicinal substances should be avoided. Lasalocid sodium has a selective antimicrobial activity against Gram-positive bacterial species while many Enterobacteriaceae are naturally resistant. Induction of resistance and/or cross-resistance was not observed in experimental conditions. Lasalocid A sodium is not genotoxic and not carcinogenic. The newly conducted cardiovascular study in dogs indicated an acute no-observed-adverse-effect level (NOAEL) of 1 mg/kg body weight (bw) per day. Since this NOAEL is above the lowest NOAEL of 0.5 mg/kg bw per day previously identified in a 2-year toxicity study in rats and a developmental study in rabbits, it is concluded that this NOAEL (0.5 mg/kg bw per day) is an appropriate base for establishing an Acceptable Daily Intake of 0.005 mg lasalocid sodium/kg bw. The use of lasalocid A sodium from Avatec® 150G at the highest proposed level of 125 mg/kg complete feed is safe for the consumer. For compliance with the maximum residue limits (MRLs), a withdrawal period of 3 days is necessary. No risk for the user is expected from the use of Avatec® 150G. The use of lasalocid A sodium from Avatec® 150G in feed for chickens for fattening and chickens reared for laying up to the highest proposed dose does not pose a risk for the environment. Insufficient evidence of the anticoccidial efficacy of lasalocid A sodium was provided in chickens for fattening/reared for laying.

Efficacy of treatments with toltrazuril 7. 5% and lasalocid sodium in sheep naturally infected with Eimeria spp / Eficácia de tratamentos com toltrazuril 7, 5% e lasalocida sódica em ovinos naturalmente infectados com Eimeria spp

Abstract The objective of this study was to evaluate the efficacy of an experimental formulation of toltrazuril 7.5% + Trimix™ on a naturally acquired infection of Eimeria spp. in suckling lambs kept on pasture and, in another trial, evaluate the comparative efficacy between lasalocid and toltrazuril 7.5% + Trimix™ in newly weaned sheep under feedlot conditions that had been naturally infected with Eimeria spp. In the first experiment, 30 suckling lambs were divided into two groups: A - treated with toltrazuril 7.5% + Trimix™ and B- control. In experiment 2, 30 weaned sheep were divided into three groups: I - treated with toltrazuril 7.5% + Trimix™, II - treated with lasalocid and III - control. Treatment group A showed an efficacy of 90, 99.4 and 87.3% on days 5, 10 and 20, respectively. Treatment group I had an efficacy of 98.2, 92.6 and 94.5%, while group II had an efficacy of 72.7, 81.6 and 95.9% on days 7, 21 and 42, respectively. Eight Eimeria species were identified; E. ovinoidalis was the most common. Treatment with the toltrazuril 7.5% +Trimix ™ formulation was effective against Eimeria spp. in suckling lambs in field conditions and lambs weaned in under feedlot conditions.

Resumo O objetivo deste estudo foi avaliar a eficácia de uma formulação experimental de toltrazuril 7,5% + Trimix™ em cordeiros mantidos em pastagem com infecção naturalmente adquirida por Eimeria spp. e, em outro teste, a eficácia comparativa entre lasalocida sódica e toltrazuril 7,5% + Trimix™ em ovinos recém-desmamados, naturalmente infectados com Eimeria spp. em condições de confinamento. No primeiro experimento, 30 cordeiros lactantes foram divididos em dois grupos: A - tratados com toltrazuril 7,5% + Trimix™; e B - controle. No experimento 2, 30 ovinos desmamados foram divididos em três grupos: I - tratados com toltrazuril 7,5% + Trimix™; II - tratados com lasalocida sódica; e III - controle. O grupo A (tratado) obteve uma eficácia de 90, 99,4 e 87,3% nos dias 5, 10 e 20, respectivamente. O grupo I teve eficácia de 98,2, 92,6 e 94,5%, enquanto o grupo II teve uma eficácia de 72,7, 81,6 e 95.9% nos dias 7, 21 e 42, respectivamente. Foram identificadas oito espécies de Eimeria sendo E. ovinoidalis a mais comum. O tratamento com a formulação de toltrazuril 7,5% + Trimix™ foi eficaz contra Eimeria spp. em cordeiros em lactação em condições de campo e em ovinos desmamados em confinamento.

One-pot synthesis and cytotoxicity studies of new Mannich base derivatives of polyether antibiotic--lasalocid acid.

Seven Mannich base derivatives of polyether antibiotic Lasalocid acid (2a-2g) were synthesized and screened for their antiproliferative activity against various human cancer cell lines. A novel chemoselective one-pot synthesis of these Mannich bases was developed. Compounds 2a-2c and 2g with sterically smaller dialkylamine substituent, displayed potent antiproliferative activity (IC50: 3.2-7.3 µM), and demonstrated higher than twofold selectivity for specific type of cancer. The nature of Mannich base substituent on C-2 atom at the aromatic ring may be critical in the search for selectivity towards a particular cancer cell.

Suplemento múltiplo com ionóforos para novilhos em pasto: desempenho / Multiple supplement with ionophores for grazing steers: performance

Avaliou-se o desempenho de 25 novilhos Holandês x Zebu, castrados, com média de peso vivo inicial de 265±50 kg, sob pastejo emBrachiaria decumbens, distribuídos em cinco grupos e em cinco piquetes, segundo os tratamentos: controle - suplementação múltipla sem ionóforos (CONT); suplementação múltipla com 100mg/cab/dia de monensina (M100); suplementação múltipla com 200mg/cab/dia de monensina (M200); suplementação múltipla com 100mg/cab/dia de lasalocida (L100); suplementação múltipla com 200mg/cab/dia de lasalocida (L200). O período experimental foi de 105 dias, com rotação dos grupos nos piquetes a cada 21 dias. A suplementação foi fornecida ad libitum. A avaliação de desempenho ocorreu mediante a pesagem dos animais, em jejum de alimento e água de 14 horas, no início de cada período e término do experimento. Os animais alimentados com suplementos com ionóforos apresentaram maior ganho de peso em relação aos do controle (0,357 vs. 0,268; P = 0,0068). Entre os ionóforos, os animais alimentados com lasalocida ganharam mais peso (0,398 vs. 0,333; P=0,0175). O melhor desempenho pode ser explicado pelo maior consumo dos suplementos pelos animais alimentados com lasalocida (0,53 vs. 0,42; P<0,0001).

The performance of 25 castrated Holstein x Zebu crossbred steers averaging 265±50kgBW, grazing on Brachiaria decumbens, during dry season was evaluated. The experiment was carried out in a completed randomized design and the animals were grouped in five different paddocks with the following treatments: control - multiple supplement without ionophores (CONT); multiple supplement with 100mg of monensin/animal/day (M100); multiple supplement with 200mg of monensin/animal/day (M200); multiple supplement with 100mg of lasalocid/animal/day (L100); and multiple supplement with 200mg of lasalocid/animal/day (L200). The experimental period was 105 days, with changing groups on paddocks every 21 days. The multiple supplement was offered ad libitum. Body weight was evaluated after 14 hours of fasting. Animals fed multiple supplement with ionophores showed higher average daily weight gain than control (0.357 vs 0.268; P= 0.0068), as well as steers fed lasalocid in comparision to monensin (0.398 vs. 0.333; P= 0.0175). Animals suplemented with lasalocid had higher intakes and higher average daily gain (0.53 vs. 0.42; P<0.0001).

Suplemento múltiplo com ionóforos para novilhos em pasto: consumo, fermentação ruminal e degradabilidade in situ / Multiple supplement with ionophores for grazing steers: intake, ruminal fermentation, and in situ degradability

Avaliou-se o efeito do suplemento múltiplo com ionóforos sobre o consumo, a fermentação ruminal e a degradabilidade in situ da matéria seca da forragem. Utilizaram-se cinco novilhos Holandês x Zebu fistulados no rúmen, com peso vivo médio de 350kg, em delineamento em quadrado latino. Os tratamentos foram: suplementação múltipla sem ionóforos (CONT); suplementação múltipla com 100mg/cab/dia de monensina (M100); suplementação múltipla com 200mg/cab/dia de monensina (M200); suplementação múltipla com 100mg/cab/dia de lasalocida (L100); e suplementação múltipla com 200mg/cab/dia de lasalocida (L200). O uso de ionóforos no suplemento não influenciou o consumo de forragem, que foi, em média, 7,24kg MS/dia. A presença de ionóforos resultou em ligeiro aumento do pH ruminal em relação à ausência desses aditivos (P<0,05). Houve diferença na concentração do N-NH3 apenas para os teores de ionóforos em que 200mg/cab/dia reduziu a quantidade de N-NH3. As concentrações de acetato, propionato e butirato não foram influenciadas pela inclusão, pelo tipo ou pelos teores de ionóforos. A fração solúvel média (A) da Brachiaria decumbens foi igual a 22 por cento, e a fração insolúvel potencialmente degradável média (B) igual a 65 por cento, resultando em degradação potencial média de 87 por cento. A taxa de degradação média (c) foi de 0,03/hora. Os ionóforos não alteraram a degradação in situ da matéria seca.

The effect of multiple supplement with ionophores was evaluated on intake, ruminal fermentation, and in situ degradability of dry matter (DM) of the pasture forage. Five rumen fistulated Holstein x Zebu steers averaging 350kg of BW were used. The animals were grouped in five different paddocks under Latin Square experimental design. The treatments were multiple supplement without ionophores (CONT); multiple supplement with 100mg of monensin/animal/day (M100); multiple supplement with 200mg of monensin/animal/day (M200); multiple supplement with 100mg of lasalocid/animal/day (L100); multiple supplement with 200mg of lasalocid/animal/day (L200). The pasture intake was 7.24kg DM/day and it was not affected by ionophores. The average pH was influenced (P<0.05) by the presence of the ionophores in the supplements. There rumen N-NH3 concentration was negatively influenced by the ionophores levels in the multiple supplement. The molar concentrations of acetate, propionate, and butirate in the rumen were not affect by the presence, type, or level of ionophores. The mean soluble fraction A of Brachiaria decumbens was 22 percent, the mean potential degradable insoluble fraction (B) was 65 percent, and the degradability was 87 percent. The mean degradation rate (c) was 0.03/h. The ionophores did not affect DM in situ degradability.

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.

Desenvolvimento de novilhas de corte sob alternativas de mineralização em pastagem de azevém / Development of beef heifers under mineralization alternatives, on Italian ryegrass pasture

Foi avaliado o desenvolvimento corporal de bezerras de corte em pastagem de azevém (Lolium multiflorum Lam.), recebendo diferentes alternativas de suplementação mineral, associadas ou não à adição de ionóforo. Os tratamentos testados foram: "Sal comum": mineralização com cloreto de sódio; "Sal 40P": mineralização com sal mineral contendo 40g de fósforo por kg de produto; "Sal 40P + ionóforo": mineralização com 40g de fósforo + 2.000mg de lasalocida sódica por kg de produto. Os valores de proteína bruta, fibra em detergente neutro, Ca e P na forragem aparentemente consumida, consumo de sal, escore de condição corporal, relação peso: altura e GMD foram semelhantes (P>0,05) entre animais que receberam "Sal comum", "Sal 40P" e "Sal 40P + ionóforo". Em pastagem de azevém, sem restrição ao consumo voluntário, a lasalocida sódica adicionada ao sal mineral proporciona maior peso vivo de bezerras aos 12 meses de idade e um retorno financeiro positivo.

Body development of beef heifers grazing Italian ryegrass pasture (Lolium multiflorum Lam.) was evaluated. The animals received different alternatives of mineral supplement: associated or not to ionophore addition and the treatments were: 'Common salt': mineralization with sodium chloride; '40P salt': mineralization with mineral salt plus 40g of phosphorus per kilo of product; 40P salt + ionophore: mineralization with 40g of phosphorus plus 2000mg of lasalocid per kg of product. The values of crude protein, neutral detergent fiber and Ca and P of the apparently consumed forage, salt intake, body condition and body weight:height relation were similar (P>0.05) between animals receiving 'common salt', '40P salt' and '40P salt+ionophor'. For beef heifers grazing ryegrass pasture without intake restriction, the lasalocid associated to mineral salt provides a greater body weight at twelve months age and a positive financial.

Avaliação de ionóforos pela técnica da perda do potássio celular e produção de gases in vitro / Ionophores evaluation by intracellular potassium depletion and in vitro gas production

Dois estudos foram realizados com vacas lactantes utilizadas como unidade experimental e doadoras de líquido ruminal, sendo as populações de bactérias utilizadas para avaliar a ação de níveis crescentes de lasalocida e monensina na resistência à perda de potássio intracelular, e para produção de gases in vitro. A perda de potássio (Kmax) da lasalocida foi menor para a população de bactérias obtidas do líquido de rúmen de vacas submetidas a dietas com monensina, óleo de soja e monensina mais óleo de soja (19,4 a 25,4 por cento) quando comparada com a perda de potássio em vacas submetidas a dietas sem ionóforo e óleo de soja (30,1 por cento). O mesmo ocorreu para a perda de potássio da monensina, em que o menor valor foi de 6,5 por cento para monensina mais óleo e o maior, de 29,5 por cento, para o controle. Necessita-se de alta concentração de monensina (Kd= 2,3µM), porém baixa de lasalocida (Kd= 0,2µM) para causar a metade da perda máxima de potássio intracelular da população de bactérias do rúmen de vacas submetidas a dietas com monensina. As populações de bactérias de vacas submetidas às dietas com monensina foram sensíveis à lasalocida. As amostras incubadas com própolis produziram menor volume de gases (12,9ml/100g de MS)

Two studies were carried out with lactating cows as experimental units and ruminal fluid donors. The ruminal bacteria population was used to evaluate the action of increasing levels of lasalocid and monensin on resistance of intracellular potassium depletion and in vitro gas production intracellular depletion potassium (Kmax) of lasalocid was lower to ruminal bacteria population obtained from rumen of cows fed diets with monensin, soybean oil and monensin plus soybean oil (19.4 to 25.4 percent) when compared to cows fed with control diet (30.1 percent). The same occurred for intracellular depletion potassium (Kmax) of monensin, in which the lowest value was 6.5 percent to monensin plus soybean oil and the greatest was 29.5 percent to control. High monensin concentration (Kd= 2.3µM) and low lasalocid concentration (Kd= 0.2µM) were necessary to cause half of maximum potassium depletion in ruminal bacteria population from cows fed diet with monensin. The ruminal bacteria population from cows feed diet with monensin were sensible to lasalocid. In vitro gas production showed the lowest volume when diets were incubated with propolis (12,9ml/100g of DM)

Efeito do pH in vitro sobre a resistência de bactérias do rúmen à perda de potássio intracelular e efeito do pH e de ionóforos sobre a produção de amônia e proteína microbiana / In vitro effect of pH on resistance of ruminal bacteria to intracellular potassium depletion, and effect of pH and ionophores on ammonia and microbial protein production

Em dois estudos, o líquido ruminal de bovinos mantidos sob pastagem foi usado para incubação in vitro em diferentes meios artificiais com valores de pH 5,5 e 7,0, para avaliar a ação de níveis crescentes de monensina na resistência à perda de potássio de bactérias do rúmen e verificar o efeito de monensina e lasalocida na produção de amônia e de proteína microbiana em pH 5,5 e 7,0. O meio utilizado para determinar a perda de potássio interferiu nos valores absolutos de potássio. A concentração de monensina necessária para causar a metade da perda máxima de potássio foi de 2,77µM em pH 5,5 e 0,056µM em pH 7,0, evidenciando que as bactérias incubadas em meios com pH 5,5 foram mais resistentes à monensina que aquelas incubadas em meios com pH 7,0. Os ionóforos e a acidez do meio reduziram a produção de amônia, e não se observou interação entre eles. Os ionóforos, independente do pH, inibiram a produção de amônia em 56%. A acidez inibiu a produção de amônia em 50,5%, independente do ionóforo. Os efeitos dos ionóforos e da acidez foram aditivos quando a inibição máxima ocorreu pelo uso de ionóforos com pH baixo (75,2%). A produção de proteína microbiana foi menor quando a lasalocida estava presente no meio de cultura com baixo valor de pH.

Ruminal fluid from steers fed on pasture was incubated with artificial media at pH 5.5 and 7.0 in two experiments. In the first, the effect of monensin level on resistance of ruminal bacteria to potassium depletion was evaluated; in the second, effects of the ionophores monensin and lasalocid on ammonia and protein production were quantified. In experiment 1, culture media affected potassium level. The monensin concentration needed to cause half maximal potassium depletion was 2.77µM at pH 5.5 but was 0.056µM at pH 7.0, showing that bacteria incubated at pH 5.5 were more tolerant to monensin than those incubated at pH 7.0. Both ionophores as well as increased acidity caused decreased ammonia production. Both ionophores inhibited ammonia production by 56%, independently of pH. In cultures incubated at pH 5.5 compared to pH 7.0, ammonia production was decreased by 50.5%, independently of the ionophores. Therefore, effects of ionophores and acidity were additive, and the maximum inhibition occurred in the presence of an ionophore at low pH (75.2%). Microbial protein production was lowest when lasalocid was present in a low pH culture medium, causing inhibition of microbial growth.