Population Pharmacokinetics and Absolute Oral Bioavailability of Lasalocid after Single Intravenous and Intracrop Administration in Laying Hens.

Lasalocid sodium is a polyether carboxylic ionophore agent authorized by the EU for use as a coccidiostat in broilers, turkeys, and pullets up to 16 weeks of age, except for laying hens. However, laying hens are the most common nontarget species exposed to lasalocid sodium, mainly due to cross-contamination from feed mills. This exposure may result in potential drug residue deposition in eggs, which could potentially expose consumers to the drug. The breeds commonly used for commercial egg production in Poland are Isa Brown and Green-legged Partridge hens, which have been found to significantly differ in egg-laying performance. This variability may also affect the pharmacokinetics of lasalocid. Data on lasalocid plasma pharmacokinetics in laying hens are lacking. In this study, we aimed to determine typical population pharmacokinetic parameters, absolute oral bioavailability, and how breed may influence the pharmacokinetics of lasalocid. Twenty-layer hens of the two breeds were used in this study. Lasalocid was administered orally at a single dose of either 1 mg or 5 mg/kg body weight or intravenously at a dose of 1 mg/kg body weight, in a crossover design with a three-week washout period between study periods. Blood samples were collected for 72 h, and lasalocid concentrations were measured using high-performance liquid chromatography with fluorescence detection. A population pharmacokinetic analysis was conducted using nonlinear mixed effects modeling. Standard numerical and graphical criteria were used to select the best model, and a stepwise covariate modeling approach was used to determine any influencing factors. The best model was a three-compartment mammillary model with first-order absorption, transit compartments, and linear elimination. The estimated absolute oral bioavailability was low (36%). It was found that breed significantly influenced not only absorption but also the elimination of lasalocid. This study revealed that lasalocid absorption and elimination varied between the two breeds. This variability in pharmacokinetics may result in breed-related differences in drug residue accumulation in eggs, and ultimately, the risk associated with consumer exposure to drug residues may also vary.

In vitro comparison of the dermal penetration of three different topical formulations containing lasalocid.

BACKGROUND: Topical antimicrobial preparations are of utmost importance in treating suspected and confirmed meticillin-resistant Staphylococcus pseudintermedius (MRSP) infections due to the increasing incidence of widespread resistance to systemic antimicrobials. Lasalocid is active against MRSP in vitro and this may become an important topical antimicrobial for the treatment of canine pyoderma. HYPOTHESIS/OBJECTIVES: To determine effects of various formulation types on penetration and retention of lasalocid applied to canine skin in vitro. ANIMALS: Normal canine skin was collected from the thorax of five dogs that had been euthanized on the basis of health and/or intractable behavioural issues. METHODS: Solution, lotion and ointment containing 2% lasalocid were applied to ex vivo canine skin. Transdermal penetration was assessed for a 24 h period and retention of lasalocid was assessed at the conclusion of the study. RESULTS: The solution had significantly higher skin retention of lasalocid and proportion of applied dose retained in skin than lotion and ointment (Tukey-Kramer Honest Significant Difference test, P < 0.01). Lasalocid could not be detected in the receptor fluid of any Franz cell at any time point. CONCLUSIONS AND CLINICAL IMPORTANCE: Lasalocid was not identified in the receptor fluid of any sample, indicating that systemic absorption of the active ingredient in vivo is unlikely. Lasalocid may be useful in the treatment of MRSP infections if in vivo studies support safety and efficacy.

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)

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.

Development of Intramammary Drug Delivery Systems: Novel In Vitro Release Method.

The aim of this study was to develop an in vitro release model for intramammary drug delivery system (IMDS) evaluation. This study was the first to establish an in vitro-in vivo correlation with investigation of an IMDS containing lasalocid. Three different methods including the standard United States Pharmacopeia dissolution method with apparatus 2, a modified United States Pharmacopeia method using a dialysis bag, or a specifically designed enhancer cell system, were assessed for the release study. Full cream milk and water were selected as the release media. In vivo evaluation was carried out by administering lasalocid IMDS into the udder of lactating Holstein dairy cows. Milk samples were collected and analyzed at selected time points after treatment. Dissolution data were fitted to various kinetic models. The results indicated that the release rate of lasalocid from IMDS was controlled by factors other than diffusion, which could include the sedimentation of lasalocid to the interface and the wetting of lasalocid particles by water at the interface of oil in the formulation and release media. The results obtained in vivo and in vitro were consistent. The in vitro assessment supports formulation design for early stage development and potentially for in vivo performance analysis.

Development of intramammary delivery systems containing lasalocid for the treatment of bovine mastitis: impact of solubility improvement on safety, efficacy, and milk distribution in dairy cattle.

BACKGROUND: Mastitis is a major disease of dairy cattle. Given the recent emergence of methicillin-resistant Staphylococcus aureus as a cause of bovine mastitis, new intramammary (IMA) treatments are urgently required. Lasalocid, a member of the polyether ionophore class of antimicrobial agents, has not been previously administered to cows by the IMA route and has favorable characteristics for development as a mastitis treatment. This study aimed to develop an IMA drug delivery system (IMDS) of lasalocid for the treatment of bovine mastitis. METHODS: Minimum inhibitory concentrations (MICs) were determined applying the procedures recommended by the Clinical and Laboratory Standards Institute. Solid dispersions (SDs) of lasalocid were prepared and characterized using differential scanning calorimetry and Fourier transform infrared spectroscopy. IMDSs containing lasalocid of micronized, nano-sized, or as SD form were tested for their IMA safety in cows. Therapeutic efficacy of lasalocid IMDSs was tested in a bovine model involving experimental IMA challenge with the mastitis pathogen Streptococcus uberis. RESULTS: Lasalocid demonstrated antimicrobial activity against the major Gram-positive mastitis pathogens including S. aureus (MIC range 0.5-8 µg/mL). The solubility test confirmed limited, ion-strength-dependent water solubility of lasalocid. A kinetic solubility study showed that SDs effectively enhanced water solubility of lasalocid (21-35-fold). Polyvinylpyrrolidone (PVP)-lasalocid SD caused minimum mammary irritation in treated cows and exhibited faster distribution in milk than either nano or microsized lasalocid. IMDSs with PVP-lasalocid SD provided effective treatment with a higher mastitis clinical and microbiological cure rate (66.7%) compared to cloxacillin (62.5%). CONCLUSION: Lasalocid SD IMDS provided high cure rates and effectiveness in treating bovine mastitis with acceptable safety in treated cows.

Effects of lasalocid and intermittent feeding of chlortetracycline on the growth of prepubertal dairy heifers.

Forty Holstein heifers entered the 12-wk study at approximately 12 wk of age. At enrollment, heifers were blocked by birth date and assigned to 1 of 4 treatments: (1) carrier (30 g; control); (2) lasalocid + carrier (1 mg/kg of body weight; L); (3) chlortetracycline + carrier (22 mg/kg of body weight; CTC); (4) L + CTC + carrier (CTCL). Heifers on CTC and CTCL were provided treatment Monday through Friday and carrier only on Saturday and Sunday. These heifers were provided their respective treatment during wk 1 to 4, 6, and 10; wk 5, 7 to 9, and 11 to 12 heifers were provided the nonmedicated carrier. Heifers were individually fed a total mixed ration with treatments top-dressed at 1200 h daily. Dry matter intake was monitored for each heifer and feed provided was adjusted according to individual intakes. Skeletal measurements were taken weekly and blood samples were obtained every Monday, Wednesday, and Friday. Blood samples were analyzed for thyroxine concentration via radial immunoassay. Heifers supplemented with L had lower average daily gain , overall body weight gain, and trends for lower daily body length gain and overall girth gain compared with CTC heifers, but similar to control and CTCL heifers. Heifers fed L had lower hip height gain and overall hip height gain compared with CTCL heifers, but similar to control and CTC heifers. Heifers fed L had lower overall withers height gain compared with control heifers, but similar to CTC and CTCL heifers. No treatment effect on thyroxine concentrations was observed. These data indicate that L did not increase growth. Results from this experiment indicate that supplementing heifers with L was not beneficial and no benefits to supplementing heifers with CTC or the combination of CTC and L were evident compared with control heifers. Heifers in this study experienced minimal health problems and were regarded to be under low stress levels. Supplementing CTC and L may be beneficial to growing heifers under conditions where disease exposure and stressors are greater.

Transfer of the coccidiostats monensin and lasalocid from feed at cross-contamination levels to whole egg, egg white and egg yolk.

Recent legislation has addressed the unavoidable carry-over of coccidiostats and histomonostats in feed, which may lead to the presence of residues of these compounds in eggs. In this study, laying hens received cross-contaminated feed at a ratio of 2.5%, 5% and 10% of the therapeutic dose of monensin and lasalocid for broilers. The eggs were collected during the treatment and depletion period and were analysed using liquid chromatography-tandem mass spectrometry. The different egg matrices were separated and analysed during the plateau phase. High lasalocid concentrations, which exceeded the maximum residue level, and low monensin concentrations were found in whole egg. Plateau levels were reached at days 7-9 for lasalocid and at days 3-5 for monensin. For lasalocid, the highest concentrations were measured in egg yolk; residue concentrations in egg white were very low.

Tissue Doppler imaging and 2-dimensional speckle tracking of left ventricular function in horses exposed to lasalocid.

BACKGROUND: Tissue Doppler imaging (TDI) and 2-dimensional speckle tracking (2DST) can quantify left ventricular (LV) function in horses. OBJECTIVES: To evaluate LV function by TDI and 2DST in horses with myocardial dysfunction after accidental ionophore intoxication. ANIMALS: Sixty-seven horses exposed to lasalocid in feed. METHODS: Prospective study. Horses were included in the study if a full cardiac examination was performed, consisting of determination of cardiac troponin I (cTnI), electrocardiography, and echocardiography. By TDI, radial systolic velocity and strain were measured. By 2DST, circumferential (SC) and radial (SR) strain at papillary muscle and chordal level and longitudinal (SL) strain were measured. RESULTS: Twenty horses showed signs of myocardial injury. Forty-nine examinations were performed on these horses between day 30 and 490 after suspected onset of exposure. Five horses had increased cTnI and ventricular tachycardia and 15 had increased cTnI without ventricular tachycardia. Horses with mild myocardial damage showed few significant differences compared with a control group. Horses with severe myocardial damage showed severely decreased TDI, 2DST and fractional shortening measurements (P < .05), indicating impaired LV function. Long-term follow-up of 2 surviving horses demonstrated full recovery in 1 horse and permanent myocardial fibrosis in the other. The lowest measurements per horse (n = 20) for all TDI measurements, SL, SR at chordal level, and FS correlated significantly with maximal cTnI (P < .05). Over all examinations (n = 49), TDI and 2DST measurements correlated well with FS (P < .05). CONCLUSIONS AND CLINICAL IMPORTANCE: The TDI and 2DST measurements allowed accurate detection and quantification of LV dysfunction in horses exposed to lasalocid.

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)

Effects of lasalocid or monensin supplementation on digestion, ruminal fermentation, blood metabolites, and milk production of lactating dairy cows.

Six ruminally fistulated midlactating multiparous Holstein cows were used in a double 3 x 3 Latin square design (35-d periods) to study the effects of lasalocid (LAS) and monensin (MON) supplemented at 24 mg/ kg of dry matter on digestion, ruminal fermentation, blood metabolites, and milk production. Cows were blocked according to milk production and fed a red clover silage-based total mixed ration (17.8% crude protein) without supplementation or supplemented with LAS or MON. Daily dry matter intake, milk production, and milk fat and protein concentrations were similar among treatments and averaged 23.5 kg, 36.6 kg, 3.36%, and 3.38%, respectively. Rumen lipogenic:glucogenic volatile fatty acids and NH(3)-N concentration were lower, and apparent digestibility of dry matter, organic matter, crude protein, and gross energy were higher with than without ionophore supplementation. Compared with LAS, MON increased concentrations of plasma urea-N and milk urea-N, and excretion of urinary urea-N and total N. Monensin also decreased N retention and tended to reduce plasma concentration of nonessential AA in comparison with LAS. Both ionophores reduced daily fecal excretion of N by 13 g compared with the control, but MON increased daily losses of urinary N by 36 g compared with LAS. Results from this study suggest that postabsorptive metabolism of N might be altered by the type of ionophore fed.

Changes in net portal nutrient flux in response to weaning transition and ionophore supplementation in dairy calves.

Dairy calf weaning is associated with ketone concentrations that exceed the levels occurring in adults, and weaning represents a potential energy loss that may be mitigated by ionophore supplementation. To assess the effects of weaning and ionophore supplementation on net nutrient flux across portal-drained viscera (PDV) tissues in dairy calves, concentrations of glucose, acetoacetate (ACAC), beta-hydroxybutyrate (BHBA), nonesterified fatty acids, volatile fatty acids, lactate, pyruvate, insulin, and glucagon and PDV flux rates were determined in Jersey bull calves (n = 19) at 35, 56, 84, and 112 d of age. Calves were randomly assigned at birth to either a commercial pelleted starter without (CON) or with lasalocid (TRT; 83 mg/kg of dry matter). Calves were fed only milk replacer from d 3 to 34 (d 3 to 20 = 454 g/d; d 21 to 34 = 568 g/d). After blood sampling on d 35, calves received replacer (d 35 to 41 = 454 g/d; d 42 to 48 = 227 g/d) and had free access to the CON or TRT starter, and from d 49 to 112 they received CON or TRT ad libitum. Catheters were implanted in the portal vein and in the mesenteric vein and artery between d 21 and 28. Blood flow was measured by continuous infusion of p-aminohippurate into the mesenteric vein. Six serial samples were taken at 30-min intervals from the arterial and portal vein catheters simultaneously. Portal blood flow increased with age but did not differ between CON and TRT calves. Glucose was released preweaning and was extracted postweaning by PDV, but was not affected by ionophore. The portal flux of nonesterified fatty acids was not different from zero during any of the 4 sample ages. Fluxes of ACAC and BHBA in CON and TRT calves went from no measurable flux preweaning to a postweaning PDV release that peaked at d 84, but the d-84 release of ACAC and BHBA was lower in TRT calves. The portal flux of volatile fatty acids increased with age, and PDV release of both butyrate and propionate was lower at d 84 in TRT than in CON calves. However, TRT calves had a greater PDV release of lactate on d 84, partially compensating for the lower release of propionate. Glucagon was greater in CON than in TRT calves at d 84 and could be a response to the elevated ketogenesis observed in CON calves during this period. Changes in the metabolic profile and nutrient flux of transition calves were demonstrated in response to weaning and ionophore supplementation. Inclusion of an ionophore appeared to moderate alimentary output at a postweaning period (d 84) at which ketone concentrations have the potential to exceed the whole animal capacity for utilization.

Effects of weaning and ionophore supplementation on selected blood metabolites and growth in dairy calves.

Dairy calf weaning results in blood ketone concentrations in excess of mature rates of use and can result in excretion of ketones in urine representing a loss of energy. Lasalocid is frequently supplemented as an anticoccidial agent in calf starters; however, in mature ruminants it is known to alter molar ratios of ruminal volatile fatty acids (VFA). Effects of weaning transition and postweaning ionophore supplementation on body weight, dry matter (DM) intake, average daily gain (ADG), and blood concentrations of glucose, acetoacetate, beta-hydroxybutyrate (BHBA), lactate, pyruvate, nonesterified fatty acids (NEFA), VFA, insulin, and glucagon were examined using Jersey bull calves (n = 24) over 16 wk. Calves were blocked into groups of 2 according to birth date and weight and randomly assigned to receive either a commercial pelleted starter (control), or the same diet containing lasalocid (TRT; 83 mg/kg of DM). Calves were fed milk replacer from d 3 to 34 (d 3 to 20 = 454 g/d at 12% solids; d 21 to 34 = 568 g/d at 15% solids), from d 35 to 48 calves received both replacer (d 35 to 41 = 454 g/d; d 42 to 38 = 227 g/d) and free access to control or TRT starter, and from d 49 to 112 received ad libitum control or TRT. Body weight and jugular blood metabolite concentrations were measured and recorded weekly. Postweaning DM intake, average daily gain, and feed:gain did not differ between control and TRT calves. Glucose and NEFA concentrations did not differ between control and TRT, but declined with age. Insulin and glucagon concentrations did not differ between control and TRT, but glucagon concentrations increased with weaning. Total VFA significantly increased following introduction of solid feed at d 35 in both groups with an apparent 1-wk lag in TRT VFA increases compared with control. Jugular acetate and butyrate concentrations were greater in control calves than TRT calves during wk 7. Propionate concentrations did not differ between control and TRT at any time following weaning. Blood BHBA concentrations were greater in control than TRT during wk 8 and 9. Thus, consumption of starter supplemented with lasalocid delayed peak acetate and butyrate and lowered peak BHBA concentrations. However, supplementation at concentrations currently recommended for control of coccidiosis did not appear to be sufficient to enhance growth or efficiency during the wk 7 to 16 postweaning interval for this sample size.

Efficacy of ionophores in cattle diets for mitigation of enteric methane.

Use of ionophores in cattle diets has been proposed as a strategy for mitigation of enteric CH4 emissions. Short- and long-term effects of feeding a single ionophore (monensin) or rotation of 2 ionophores (monensin and lasalocid) on enteric CH4 emissions were evaluated in 36 Angus yearling steers (328 +/- 24.9 kg of BW) over a 16-wk period. Steers were randomly assigned to 6 dietary treatments of 6 steers each. The 6 diets were low-concentrate without ionophore supplementation, low-concentrate with monensin supplementation, low-concentrate with a 2-wk rotation of monensin and lasalocid supplementation, high-concentrate without ionophore supplementation, high-concentrate with monensin supplementation, and high-concentrate with a 2-wk rotation of monensin and lasalocid supplementation. Daily enteric CH4 emissions, as measured using the SF(6) tracer gas technique, ranged from 54.7 to 369.3 L/steer daily. Supplementing ionophores decreased (P < 0.05) enteric CH4 emissions, expressed as liters per kilogram of DMI or percentage of GE intake, by 30% for the first 2 wk and by 27% for the first 4 wk, for cattle receiving the high-concentrate and low-concentrate diets, respectively. Cattle fed a rotation of ionophores did not (P > 0.05) exhibit a greater decrease and did not (P > 0.05) have a longer period of depressed enteric CH4 emissions compared with cattle receiving monensin only. Ionophore supplementation did not (P > 0.05) alter total ruminal fluid VFA concentration; however, the acetate:propionate ratio and ammonia-N concentration in ruminal fluid were decreased (P < 0.001) from the time that ionophores were introduced to the time they were removed from the diets. Both monensin and the rotation of monensin and lasalocid decreased (P < 0.001) total ciliate protozoal populations by 82.5% in the first 2 wk and by 76.8% in the first 4 wk during which they were supplemented in the high-concentrate and low-concentrate diets, respectively. Original ciliate protozoal populations were restored by the fourth and sixth week of supplementation when cattle were fed the high- or low-concentrate diets, respectively. No significant change was observed thereafter. These data suggest that the effects of ionophores on enteric CH(4) production are related to ciliate protozoal populations and that ciliate protozoal populations can adapt to the ionophores present in either low- or high-concentrate diets. Rotation of monensin and lasalocid did not (P > 0.05) prevent ciliate protozoal adaptation to ionophores.

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.

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.

Production and health of pasture-fed dairy cattle following oral treatment with the ionophore lasalocid.

The aim of the study was to evaluate the effect of feeding the ionophore lasalocid on the productivity and health of seasonally calving, pasture-fed dairy cows. Dairy cows (n = 1020) from 4 herds were enrolled in a split-herd, prospective intervention study. Cows were blocked by breed and age, ranked on previous production, and then assigned to 2 treatment groups. Treatment cows were each exposed to 300 to 350 mg of lasalocid/d commencing 3 wk before and ending 18 wk after the start of the seasonal calving period. Milk production was determined on 3 occasions for each cow at approximately monthly intervals (herd tests 1 to 3), body condition score was determined fortnightly, and all disease occurrences were recorded. Lasalocid treatment increased milk volume milk protein and milk fat production by approximately 2%, without altering milk composition. Fewer lasalocid-treated cows than control cows (7.3 vs. 11.6%, respectively) were diagnosed with clinical mastitis. Lasalocid treatment of pasture-fed dairy cows resulted in reduced mastitis incidence and increased milk production without changes in body condition or negative effects on metabolic processes as monitored by metabolite concentrations.

An alternative to antibiotic-based drugs in feed for enhancing performance of broilers grown on Eimeria spp.-infected litter.

Three trials were conducted to evaluate the effects of lasalocid, an anticoccidial feed additive (90.7 kg/ton); bacitracin, a growth-promoter (50 g/ton); and yeast culture residue (YCR) (1 kg/ton) on the performance of broiler chicks reared to 42 d of age on recycled litter. Recycled litter consisted of pine wood shavings containing droppings from chicks infected with 3 select strains of coccidia (Eimeria tenella, Eimeria maxima, and Eimeria acervulina). Response variables (BW, intestinal tract and litter coliform counts, cecal and liver relative weights, and litter moisture content) were recorded biweekly. Mean BW of chicks fed the diet supplemented with YCR was higher than that of the controls (P < 0.05) and comparable to that of the lasalocid-treated birds in all 3 trials. Mean BW of chicks in all treatment groups decreased uniformly as the litter aged and moisture content increased. The mean intestinal coliform population from YCR-treated chicks was lower (P < 0.05) than those of the control and lasalocid populations. The coliform count was consistently lower than that in chicks on a bacitracin-supplemented diet. Coliform counts from the control and lasalocid-treated birds did not differ. The litter coliform counts increased with increased use of the litter. Cecal and liver relative weights calculated from the chicks in trial 3 showed that only the liver was significantly affected by treatments. YCR appeared to be a viable alternative to bacitracin and lasalocid medication in enhancing growth of broiler chicks reared on recycled litter.

Effect of lonophore supplementation on selected serum constituents of sheep consuming locoweed.

The effects of ionophore supplementation on selected serum constituents of sheep consuming locoweed were investigated. Sixteen sheep were allotted by weight to a 2x2 factorial arrangement of treatments: 1) no locoweed, no lasalocid, 2) no locoweed, 0.75 mg lasalocid/kg BW, 3) 0.5 mg swainsonine/kg BW, no lasalocid, 4) 0.5 mg swainsonine/kg BW, 0.75 mg lasalocid/kg BW. Swainsonine was provided by locoweed (Oxytropissericea), and sheep were fed a blue grama based diet at 2.5% BW for a 35 d treatment period. Diets were formulated to be isocaloric and isonitrogenous. Blood samples were collected on d 1, 7,14, 21, 31 and 35 to determine serum swainsonine concentration, alkaline phosphatase, total iron, aspartate aminotransferase, g-glutamyltransferase, and lactate dehydrogenase activity and total cholesterol, and triglyceride concentrations. No lasalocid by locoweed interaction (P > 0.4) was noted for any response variable measured. Average daily gains (P = 0.4) and orts (P = 0.7) were not affected by the treatments. No lasalocid treatment (P = 0.7) or day (P = 0.1) effect of serum swainsonine was observed. A locoweed by day interaction (P < 0.0001) of serum alkaline phosphatase was detected. Alkaline phosphatase levels were elevated (P < 0.01) for locoweed treated sheep at 24 h following initial exposure and remained elevated throughout the trail. Total iron was suppressed (P < 0.08) in locoweed fed sheep. A day effect (P < 0.02) was observed for serum iron. However, no linear, quadratic, or cubic effects of day were noted (P >0.2). A locoweed by day interaction (P < 0.0001) of serum aspartate aminotransferase and g-glutamyltransferase was detected. Aspartate aminotransferase levels were elevated (P < 0.0001) by d 7 for locoweed treated animals and remained elevated throughout the trial. g--Glutamyltransferase levels were suppressed (P < 0.0001) by day 7 for locoweed treated animals and remained suppressed throughout the trial. A locoweed by day interaction (P = 0.06) of serum cholesterol was detected. However, no linear, quadratic, or cubic effects of day were detected (P = 0.2). Lasalocid treatment had no effect on any serum constituent measured. Use of lasalocid in grazing animals should not increase the likelihood of locoweed intoxication.

Secretion of the trefoil factor TFF3 from the isolated vascularly perfused rat colon.

The trefoil factor TFF3 is a peptide predominantly produced by mucus-secreting cells in the small and large intestines. It has been implicated in intestinal protection and repair. The mechanisms that govern TFF3 secretion are poorly understood. The aim of this study was, therefore, to evaluate the influence of neurotransmitters, hormonal peptides and mediators of inflammation on the release of TFF3. For this purpose, an isolated vascularly perfused rat colon preparation was used. After a bolus administration of 1 ml isotonic saline into the lumen, TFF3 secretion was induced by a 30-min intra-arterial infusion of the compounds to be tested. TFF3 was evaluated in the luminal effluent using a newly developed radioimmunoassay. TFF3 was barely detected in crude luminal samples. In contrast, dithiothreitol (DTT) treatment of the effluent revealed TFF3 immunoreactivity, which amounted to about 0.3 pmol min(-1) cm(-1) in the basal state. Gel chromatography of DTT-treated luminal samples revealed a single peak that co-eluted with the monomeric form of TFF3. TFF3 was not detected in the portal effluent. Bethanechol (10(-6)-10(-4) M), vasoactive intestinal peptide (VIP, 10(-8)-10(-7) M) or bombesin (10(-8)-10(-7) M) induced a dose-dependent release of TFF3. In contrast, substance P evoked a modest release of TFF3, whereas calcitonin gene-related peptide (CGRP), somatostatin, neurotensin or peptide YY (PYY) did not modify TFF3 secretion. The degranulator compound bromolasalocid, 16,16-dimethyl PGE2 (dmPGE2) or interleukin-1-beta (IL-1-beta) also evoked a marked release of TFF3. In conclusion, TFF3 in the colonic effluent is present in a complex. This association presumably involves a disulfide bond. Additionally, the present results suggest a role for enteric nervous system and resident immune cells in mediation of colonic TFF3 secretion.