RESUMO
Adding ionophores to ruminant diets is a strategy to manipulate ruminal fermentation and improve milk yield. This study evaluates the effects of narasin supply to lactating ewes on dry matter intake (DMI), milk yield and composition, and performance of the lambs. Thirty lactating Santa Inês and Santa Inês × Dorper ewes fed a basal diet containing 50 % coastcross hay and 50 % concentrate were randomly assigned to two treatments: control (CON; without ionophores) or NAR (addition of 13 mg narasin kg-1 DM). From the 2nd to 10th week of lactation, DMI of ewes was determined, and once a week, their milk production and composition was measured over a 3-h interval. At the 10th week of lactation, lambs were weaned and their average daily gain (ADG) and starter DMI continued to be evaluated for two more weeks. Narasin supply did not affect weight and DMI of ewes. Ewes fed NAR had greater feed efficiency for milk production and displayed tendency for higher milk yield. Narasin supply reduced milk protein levels, but it did not affect other milk component levels. Ewes fed NAR had greater production of milk urea nitrogen and showed tendency for higher production of fat and total solids. Starter DMI of lambs was not affected by treatments; however, there was a tendency for greater weaning weight for NAR lambs. At the end of experiment, no differences were observed in the performance of lambs. The supply of 13 mg narasin kg-1 to lactating ewes improved milk yield efficiency and tended to increase the weaning weight of their lambs.(AU)
Assuntos
Animais , Feminino , Ovinos/fisiologia , Coccidiostáticos/química , Leite/fisiologia , Animais Lactentes/fisiologia , Lactação , Ionóforos/análiseRESUMO
The diphenylurea 4,4'-dinitrocarbanilide (DNC) is the residue of concern left in edible tissues of broilers fed diets containing the anticoccidial nicarbazin. When chicken meat is submitted to thermal processing, p-nitroaniline (p-NA) is expected from DNC degradation. This work aimed at evaluating whether thermal processing of DNC-containing chicken meat induces p-NA appearance. First, a hydrolysis assay was performed in aqueous solutions at 100 °C in different pH, confirming that DNC cleavage yields p-NA. Then a novel LC-MS/MS method was used to detect traces of this aromatic amine in DNC-containing chicken breast fillets subjected to cooking methods. Our evidence showed p-NA occurrence in such chicken meat samples, which corroborated results from hydrolysis assay. The p-NA appearance in fillets was rather discrete during boiling treatment, but its concentration became pronounced over time for grilling, frying, and roasting, achieving respectively 326.3, 640.0, and 456.9 µg/kg. As far as we are concerned, no other research identified degradation products from DNC residue in heat-processed chicken fillets. Therefore, this study leads to additional approaches to assess impacts on food safety.
Assuntos
Compostos de Anilina/química , Carbanilidas/química , Coccidiostáticos/química , Resíduos de Drogas/química , Carne/análise , Nicarbazina/química , Compostos de Anilina/metabolismo , Animais , Carbanilidas/metabolismo , Galinhas/metabolismo , Coccidiostáticos/metabolismo , Culinária , Resíduos de Drogas/metabolismo , Temperatura Alta , Nicarbazina/metabolismo , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Avian coccidiosis is a disease caused worldwide by several species of parasite Eimeria that causes significant economic losses. This disease affects chickens development and production, that most of times is controlled with anticoccidial drugs. Although efforts have been made to address this disease, they have been made to control Eimeria sporozoites, although enteric stages are often vulnerable, however; the parasite oocyst remains a problem that must be controlled, as it has a resistant structure that facilitates dispersion. Despite some commercial products based on chemical compounds have been developed as disinfectants that destroy oocysts, the solution of the problem remains to be solved. RESULTS: In this work, we assessed in vitro anticoccidial activity of a compound(s) secreted by yeast isolated in oocysts suspension from infected chickens. The yeast was molecularly identified as Meyerozyma guilliermondii, and its anticoccidial activity against Eimeria tenella oocysts was assessed. Here, we report the damage to oocysts walls caused by M. guilliermondii culture, supernatant, supernatant extract and intracellular proteins. In all cases, a significant decreased of oocysts was observed. CONCLUSIONS: The yeast Meyerozyma guilliermondii secretes a compound with anticoccidial activity and also has a compound of protein nature that damages the resistant structure of oocyst, showing the potential of this yeast and its products as a feasible method of coccidiosis control.
Assuntos
Coccidiose/veterinária , Coccidiostáticos/química , Coccidiostáticos/farmacologia , Eimeria/efeitos dos fármacos , Leveduras/classificação , Leveduras/metabolismo , Animais , Galinhas , Coccidiose/prevenção & controle , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Oocistos/efeitos dos fármacos , Filogenia , Reação em Cadeia da Polimerase , RNA Fúngico/genética , RNA Ribossômico 18S/genéticaRESUMO
Toxoplasma gondii, the causative agent of toxoplasmosis, is an obligate intracellular protozoan that can infect a wide range of vertebrate cells. Here, we describe the cytotoxic effects of the dinuclear iron compound [Fe(HPCINOL)(SO4)]2-µ-oxo, in which HPCINOL is the ligand 1-(bis-pyridin-2-ylmethyl-amino)-3-chloropropan-2-ol, on T. gondii infecting LLC-MK2 host cells. This compound was not toxic to LLC-MK2 cells at concentrations of up to 200 µM but was very active against the parasite, with a 50% inhibitory concentration (IC50) of 3.6 µM after 48 h of treatment. Cyst formation was observed after treatment, as indicated by the appearance of a cyst wall, Dolichos biflorus lectin staining, and scanning and transmission electron microscopy characteristics. Ultrastructural changes were also seen in T. gondii, including membrane blebs and clefts in the cytoplasm, with inclusions similar to amylopectin granules, which are typically found in bradyzoites. An analysis of the cell death pathways in the parasite revealed that the compound caused a combination of apoptosis and autophagy. Fluorescence assays demonstrated that the redox environment in the LLC-MK2 cells becomes oxidant in the presence of the iron compound. Furthermore, a reduction in superoxide dismutase and catalase activities in the treated parasites and the presence of reactive oxygen species within the parasitophorous vacuoles were observed, indicating an impaired protozoan response against these radicals. These findings suggest that this compound disturbs the redox equilibrium of T. gondii, inducing cystogenesis and parasite death.
Assuntos
Antioxidantes/metabolismo , Coccidiostáticos/farmacologia , Inibidores Enzimáticos/farmacologia , Compostos Férricos/farmacologia , Toxoplasma/efeitos dos fármacos , Animais , Catalase/antagonistas & inibidores , Catalase/metabolismo , Linhagem Celular , Coccidiostáticos/química , Inibidores Enzimáticos/química , Compostos Férricos/química , Macaca mulatta , Microscopia Eletrônica de Transmissão , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Toxoplasma/crescimento & desenvolvimento , Toxoplasma/metabolismoRESUMO
Here we evaluate the effects of BpLec, a C-type lectin isolated from Bothrops pauloensis snake venom, on Toxoplasma gondii parasitism. BpLec (0.195-12.5 µg/mL) did not interfere with HeLa (host cell) viability by MTT assay, whereas higher doses decreased viability and changed HeLa morphology. In addition, the host cell treatment before infection did not influence adhesion and proliferation indexes. BpLec did not alter T. gondii tachyzoite viability, as carried out by trypan blue exclusion, but decreased both adhesion and parasite replication, when tachyzoites were treated before infection. Galactose (0.4 M) inhibited the BpLec effect on adhesion assays, suggesting that BpLec probably recognize some glycoconjugate from T. gondii membrane. Additionally, we performed cytokine measurements from supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BpLec. MIF and IL-6 productions by HeLa cells were increased by BpLec treatment. Also, TGF-ß1 secretion was diminished post-infection, although this effect was not dependent on BpLec treatment. Taken together, our results show that BpLec is capable of reducing T. gondii parasitism after tachyzoite treatment and may represent an interesting tool in the search for parasite antigens involved in these processes.
Assuntos
Bothrops/metabolismo , Coccidiostáticos/química , Coccidiostáticos/farmacologia , Lectinas Tipo C/química , Toxoplasma/efeitos dos fármacos , Peçonhas/química , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Coccidiostáticos/isolamento & purificação , Citocinas/metabolismo , Células HeLa , Humanos , Lectinas Tipo C/isolamento & purificaçãoRESUMO
The efficacy of three amino-terpenyl naphthoquinones and the alkaloid liriodenine were examined against tachyzoites and tissues cysts of the RH and EGS strains, respectively. Monolayers of 2C4 fibroblasts infected with tachyzoites of the RH strain were incubated with different concentrations of the compounds for 48 h. Specifically, 7-(4-methyl-3-pentenyl)-2-pyrrolidine-[1,4]-naphthoquinone (QUI-5), 6-(4-methyl-3-pentenyl)-2-pyrrolidine-[1,4]-naphthoquinone (QUI-6), 6-(4-methylpentyl)-2-pyrrolidine-[1,4]-naphthoquinone (QUI-11), and 8 h-benzo[g]-1,3-benzodioxolo[6,5,4-de]quinolin-8-one,9Cl-1,2-methylene dioxiaporfina (liriodenine) inhibited intracellular replication of T. gondii. The IC(50) values obtained for compounds QUI-5 and QUI-6 were 69.35 and 172.81 µM (i.e., 21.4 and 53.4 µg/mL), respectively. The naphthoquinone QUI-11 and liriodenine significantly inhibited intracellular replication of T. gondii. The IC(50) values obtained with these experiments were 0.32 and 0.07 µM (i.e., 0.1 and 0.02 µg/mL), respectively. Compounds QUI-5, QUI-6, QUI-11 and liriodenine demonstrated lower toxicity for 2C4 fibroblasts compared to atovaquone. In addition, cysts isolated from the brains of mice chronically infected with the EGS strain were exposed to the compounds. Infectivity of the cysts after incubation with the compounds was assessed by infection of mice. The data obtained showed that in vitro incubation with QUI-6, QUI-11 and liriodenine inhibited the infectivity of the bradyzoites. This activity was time- and concentration-dependent.
Assuntos
Aporfinas/farmacologia , Coccidiostáticos/farmacologia , Fibroblastos/parasitologia , Naftoquinonas/farmacologia , Toxoplasma/efeitos dos fármacos , Animais , Antimaláricos/química , Antimaláricos/farmacologia , Aporfinas/química , Atovaquona/química , Atovaquona/farmacologia , Células Cultivadas , Coccidiostáticos/química , Feminino , Fibroblastos/efeitos dos fármacos , Prepúcio do Pênis/citologia , Humanos , Concentração Inibidora 50 , Masculino , Camundongos , Naftoquinonas/química , Relação Estrutura-Atividade , Sulfadiazina/química , Sulfadiazina/farmacologia , Toxoplasma/patogenicidade , Toxoplasmose Animal/tratamento farmacológico , Toxoplasmose Animal/parasitologiaRESUMO
Previous studies from our group have demonstrated the high susceptibility of Toxoplasma gondii tachyzoites to the sterol analogues 22,26-azasterol and 24,25-(R,S)-epiminolanosterol. In this work we present data on testing in vitro three novel azasterols as potential agents for the treatment of toxoplasmosis. The three compounds inhibited parasite growth at micromolar concentrations, in a dose-dependent manner. Electron microscopy analysis of intracellular tachyzoites after treatment with the most effective compound showed drastic mitochondrion swelling associated with the appearance of an electron-lucent matrix and disrupted cristae. Parasite lysis also took place. The appearance of electron dense cytoplasmic structures similar to amylopectin granules distributed throughout the parasite suggests that azasterols might be inducing differentiation of those tachyzoites which were not lysed to the bradyzoite stage.
Assuntos
Coccidiostáticos/farmacologia , Esteróis/farmacologia , Toxoplasma/efeitos dos fármacos , Animais , Coccidiostáticos/química , Estrutura Molecular , Esteróis/químicaRESUMO
The aim of this study was to evaluate the anticoccidial efficacy of a product containing coumestans from Eclipta alba. Experimental conditions were set up as to reproduce the environment conditions for husbandry adopted in commercial broiler farms. Broilers were raised in broiler chicken shed provided with feeders, drinkers, illumination and temperature control systems and floor covering to afford an adequate nourishing environment. Male Cobb broilers (240) were assigned to four experimental groups being each experimental group set apart in rice straw-covered shed isolated with wire mesh. One-day-old broilers were reared in a coccidian-free environment with ad libitum supply of filtered water and freely available standard feed, from the 1st to the 35th day of life. The T1 group received standard feed (negative control); T2 was treated with standard feed supplemented with 66 ppm of salinomycin (positive control); groups T3 and T4 had standard feed supplemented with the ethyl acetate fraction from methanolic extract of E. alba aerial parts, which contains the coumestans WL and DWL (120 and 180 ppm, respectively). The chicken broilers were individually infected with 2 × 104 oocysts of Eimeria tenella when they were 14 days old and were monitored weekly to evaluate zootechnical parameters such as weight gain and food conversion ratio. Counting of coccidial oocyst in chicken feces was assessed from random samples, from the 21st to 28th days of life, which corresponded to 7-14 days after the infection. Five chickens selected at random from each experimental group were subsequently euthanized at 21, 28 or 35 days of life to determine the lesion score in the cecal region and to excise a cecum portion for histopathological evaluation. The group treated with coumestans from E. alba presented an average weight gain and food conversion ratio higher than the negative control group and similar to the mean value of the positive control group. Coumestan-treated groups showed a significant decrease in the oocyst counting since the 21 th day of life and displayed a reduced number of macroscopic lesions. Histopathological evaluations of cecum fragments showed that both treatments induced the migration of defense cells at the site of infection. A severe destruction of the cecal lining was found in the intestinal tract of broilers fed with a coumestans dose of 180 ppm. Overall, our results validate the use of a phytotherapy containing E. alba coumestans at a dose of 120 ppm as a therapeutic or prophylactic agent against avian coccidiosis.
Assuntos
Galinhas , Coccidiose/veterinária , Cumarínicos/química , Eclipta/química , Eimeria tenella , Extratos Vegetais/farmacologia , Animais , Ceco/patologia , Coccidiose/parasitologia , Coccidiose/patologia , Coccidiose/prevenção & controle , Coccidiostáticos/química , Coccidiostáticos/farmacologia , Cumarínicos/farmacologia , Masculino , Extratos Vegetais/química , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/patologia , Doenças das Aves Domésticas/prevenção & controleRESUMO
The bottleneck for the complete understanding of the structure-function relationship of flexible membrane-acting peptides is its dynamics. At the same time, not only the structure but also the dynamics are the key points for their mechanism of action. Our model is PW2, a TRP-rich, cationic peptide selected from phage display libraries that shows anticoccidial activity against Eimeria acervulina. In this manuscript we used a combination of several NMR techniques to tackle these difficulties. The structural features of the membrane-acting peptide PW2 was studied in several membrane mimetic environments: we compared the structural features of PW2 in SDS and DPC micelles, that were reported earlier, with the structure properties in different lipid vesicles and the peptide free in water. We were able to unify the structural information obtained in each of these systems. The structural constraints of the peptide free in water were fundamental for the understanding of plasticity necessary for the membrane interaction. Our data suggested that the WWR sequence is the region responsible for anchoring the peptide to the interfaces, and that this same region displays some degree of conformational order in solution. For PW2, we found that affinity is related to the aromatic region, by anchoring the peptide to the membrane, and specificity is related to the N- and C-termini, which are able to accommodate in the membrane due to its plasticity.
Assuntos
Coccidiostáticos/química , Espectroscopia de Ressonância Magnética/métodos , Peptídeos/química , Membrana Celular/metabolismo , Coccidiostáticos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Bicamadas Lipídicas/metabolismo , Modelos Moleculares , Peptídeos/metabolismo , Conformação Proteica , SoluçõesRESUMO
PW2 is an anticoccidial peptide active against Eimeria acervulina and Eimeria tenella. We determined the structure of PW2 in dodecylphosphocholine micelles. The structure showed two distinct regions: an amphipathic N-terminal 3(10) helix and an aromatic region containing WWR interface-binding motif. The aromatic region acted as a scaffold of the protein in the interface and shared the same structure in both DPC and SDS micelles. N-terminal helix interacted with DPC but not with SDS interface. Chemical shift change was slow when SDS was added to PW2 in DPC and fast when DPC was added to PW2 in SDS, indicating that interaction with DPC micelles was kinetically more stable than with SDS micelles. Also, DPC interface was able to accommodate PW2, but it maintained the conformational arrangement in the aromatic region observed for SDS micelles. This behavior, which is different from that observed for other antimicrobial peptides with WWR motif, may be associated with the absence of PW2 antibacterial activity and its selectivity for Eimeria parasites.