RESUMO
Varicellovirus bovinealpha 1 (formerly bovine alphaherpesvirus type 1, BoAHV-1) is associated with several syndromes in cattle, including respiratory disease and is one of the main agents involved in the bovine respiratory disease complex (BRDC). Its infectious cycle is characterized by latent infections with sporadic virus reactivation and transmission. Although the acute disease can be prevented by the use of vaccines, specific therapeutic measures are not available. Ivermectin (IVM) is a semi-synthetic avermectin with a broad-spectrum antiparasitic activity, which has previously shown to have potential as an antiviral drug. In this study, IVM antiviral activity against BoAHV-1 was characterized in two cell lines (MDBK [Madin Darby bovine kidney] and BT [bovine turbinate]), including the measurement of intracellular drug accumulation within virus-infected cells. IVM antiviral activity was assessed at three different drug concentrations (1.25, 2.5 and 5 µM) after incubation for 24, 48 and 72 h. Slight cytotoxicity was only observed with 5 µM IVM. Even the lowest IVM dose was able to induce a significant reduction in virus titers in both cell lines. These findings indicate that the antiviral effects of IVM were evident in our experimental model within the range of concentrations achievable through therapeutic in vivo administration. Consequently, additional in vivo trials are necessary to validate the potential utility of these results in effectively managing BoAHV-1 in infected cattle.
Assuntos
Ivermectina , Varicellovirus , Animais , Bovinos , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Antiparasitários/farmacologia , Antiparasitários/uso terapêutico , Antivirais/farmacologiaRESUMO
In vitro systems are useful tools for unravelling species differences in xenobiotic metabolism.The current work aimed to validate the technique of precision-cut liver slices (PCLS) for comparative studies on xenobiotic metabolism in swine and cattle.PCLS from swine (n = 3) and cattle (n = 3) were produced using a Brendel-VitronTM Tissue Slicer and cultured for 6 h. Tissue viability was preserved throughout the whole culture period.Metabolic viability was evaluated using the anthelmintics albendazole (ABZ) and fenbendazole (FBZ) as model drugs, as well as other substrates of hepatic monooxygenases: benzydamine (BZ) N-oxygenase (FMO-dependent), and the O-dealkylations of 7-ethoxyresorufin (EROD, CYP1A1-dependent) and 7-methoxyresorufin (MROD, CYP1A2-dependent).ABZ S-oxygenation resulted 6-fold (cattle) and 13.6-fold (swine) higher (p = 0.001) compared to FBZ S-oxygenation.Similar BZ N-oxygenation and EROD activities were observed in PCLS cultures from both species. MROD was 2.5-fold higher (p = 0.033) in swine than in cattle. Similarly, ABZ S-oxygenation was 1.7-fold higher (p = 0.0002) in swine than in cattle. Conversely, a 82% higher (p = 0.0003) rate of FBZ S-oxygenation was evidenced in PCLS cultures from cattle compared to those from swine.Overall, this work shows that PCLS cultures are useful to obtain relevant information on species differences in xenobiotic metabolism.
Assuntos
Fígado , Xenobióticos , Animais , Bovinos , Xenobióticos/metabolismo , Fígado/metabolismo , SuínosRESUMO
Ivermectin (IVM) resistance in parasitic nematodes such as Haemonchus contortus has spurred a search for substances that help to recover its efficacy. One potential agent is the natural product curcumin (CUR). In this study, CUR was combined with polyvinylpyrrolidone (PVP) (CUR/PVP) to improve its solubility and biological applicability. This study determined the effect of CUR preincubation on the effective concentration 50% (EC50) of IVM in three H. contortus isolates with different susceptibilities to IVM. The IVM EC50 was determined for three H. contortus isolates with different IVM susceptibilities using the larval migration inhibition (LMI) test. The three isolates were (i) PARAISO (IVM resistant), (ii) FMVZ-UADY (IVM susceptible), and (iii) CENID-SAI INIFAP (reference IVM susceptible). The L3 of each isolate were preincubated for 3 h with one of three concentrations of CUR (µg curcumin/mL): CONC-1 (3.67), CONC-2 (5.67), or CONC-3 (8.48). Corresponding controls were performed without CUR. The EC50 of IVM was determined for each isolate after they were exposed to the different CUR concentrations. The EC50 of IVM differed between the isolates PARAISO > FMVZ-UADY > CENID-SAI INIFAP (P < 0.05). The CUR preincubation at CONC-1 did not decrease the EC50 of IVM for any of the three isolates, suggesting a hormetic effect. By contrast, CUR preincubation at CONC-2 or CONC-3 decreased the IVM EC50 for the PARAISO isolate (P < 0.05) compared with the reference isolate and reduced the EC50 of IVM for the FMVZ-UADY and CENID-SAI INIFAP isolates below the EC50 for the CENID-SAI INIFAP isolate without CUR preincubation. In conclusion, preincubation of H. contortus L3 with CUR reduced the EC50 of IVM for field isolates classified as resistant and susceptible to IVM. The CUR preincubation reduced the IVM resistance factor in the different isolates tested.
Assuntos
Anti-Helmínticos , Curcumina , Hemoncose , Haemonchus , Animais , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Curcumina/farmacologia , Curcumina/uso terapêutico , Povidona/farmacologia , Povidona/uso terapêutico , Resistência a Medicamentos , Larva , Hemoncose/tratamento farmacológico , Hemoncose/veterináriaRESUMO
Ivermectin is one of the most widely used drugs for parasite control. Previous studies have shown a reduction in the abundance and diversity of "non-target" coprophilous organisms due to the presence of ivermectin (IVM) in bovine faecal matter (FM). Due to its breadth of behavioural habits, Calliphora vicina is a suitable dipteran species to evaluate the effects of IVM in FM. The aim of this work was to evaluate the effect of five concentrations of IVM in FM (3000, 300, 100, 30, and 3 ng/g) on the development of C. vicina. The following endpoints were evaluated: survival (between the first larval stage and emergence of new adults), larval development times to pupation and pupation times to adult, and adult emergence (% sex) and LC50. Sampling was performed from larval hatching at 60 and 120 min and at 3, 4, 5, and 12 h, and every 24 h specimens were weighed until pupae were observed. Data were analysed by ANOVA using a non-parametric Kruskal-Wallis test and as a function of elapsed development time and accumulated degree hours (ADH). Mortality at 3000 and 300 ng/g was 100% and 97%, respectively. There were statistically significant delays in adult emergence time (p = 0.0216) and in the ADH (p = 0.0431) between the control group (C) and 100 ng/g. The LC50 was determined at 5.6 ng/g. These results demonstrate the lethal and sub-lethal effects of IVM on C. vicina, while highlighting the usefulness of this species as a bioindicator for ecotoxicological studies.
Assuntos
Calliphoridae , Fezes , Ivermectina , Larva , Animais , Ivermectina/farmacologia , Calliphoridae/efeitos dos fármacos , Calliphoridae/crescimento & desenvolvimento , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Fezes/parasitologia , Bovinos , Análise de Sobrevida , Pupa/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Feminino , Antiparasitários/farmacologia , Masculino , Dose Letal Mediana , Dípteros/efeitos dos fármacos , Dípteros/crescimento & desenvolvimentoRESUMO
Organophosphates (OPs), pyrethrins and fipronil, are acaricides commonly used in cattle, mainly as pour on formulations. Scant information is available on their potential interactions with hepatic xenobiotic metabolizing enzymes. This work aimed to evaluate in vitro the potential inhibitory effects of widely employed acaricides on catalytic activities mediated by hepatic cytochrome P450 (CYP) and flavin-monooxygenase (FMO) enzymes in cattle. Bovine (n = 4) liver microsomes were incubated in the absence (control assays) and in presence of different OPs (fenthion, chlorpyrifos, ethion, diazinon and dichlorvos), fipronil and cypermethrin at 0.1-100 µm. Five oxidative enzyme activities were assayed by spectrofluorimetric or HPLC methods: 7-ethoxyresorufin O-deethylase (for CYP1A1), methoxyresorufin O-demethylase (for CYP1A2), benzyloxyresorufin O-debenzylase (for CYP2B), testosterone 6-beta hydroxylase (for CYP3A) and benzydamine N-oxidase (for FMO). All acaricides, particularly phosphorothionate-containing OPs, inhibited to some extent more than one enzyme activity. The most frequent inhibitor was fenthion, which inhibited (p < .05) all enzyme activities tested (from 22% at 1 µm to 72% at 100 µm). However, low inhibitory potencies (IC50s higher than 7 µm) of all acaricides studied were observed against the catalytic activities assayed. Therefore, the risk of in vivo metabolic interactions due to inhibition of monooxygenases would be low under common husbandry conditions.
Assuntos
Acaricidas , Microssomos Hepáticos , Bovinos , Animais , Microssomos Hepáticos/metabolismo , Acaricidas/metabolismo , Acaricidas/farmacologia , Fention/metabolismo , Fention/farmacologia , Fígado/metabolismo , OxirreduçãoRESUMO
This study aimed at determining the plasma disposition kinetics of eprinomectin (EPM) and EPM excretion pattern through milk after topical administration to dairy cattle at the recommended dose of 0.5 mg/kg and at 1 and 1.5 mg/kg. A high variability in the plasma concentration profiles was observed among animals, particularly in the Cmax values, with a coefficient of variation between 39 and 53%. The Cmax and AUC values were significantly affected by the dose administered at 1.5 mg/kg. However, such differences did not seem to follow a linear pattern among treatments. These parameters did not differ among dose rates after dose normalization; nevertheless, the simulation of a linear kinetic disposition showed a mean plasma AUC value of 254 ng.d/ml instead of the observed value of 165 ng.d/ml. EPM concentration profiles in milk were significantly lower than those measured in plasma. The Cmax and AUC milk-to-plasma ratios ranged from 0.14 to 0.26 and 0.16 to 0.21, respectively (p>0.05). The low milk-to-plasma ratio of EPM accounted for a low percentage of the fraction of the administered dose excreted through milk, being significantly higher at a dose rate of 0.5 mg/kg (0.07%) of EPM than at 1.5 mg/kg (0.04%) (p<0.05). The topical administration of EPM to lactating dairy cows at higher doses than that recommended for gastrointestinal nematodes showed a milk excretion pattern with a zero milk withdrawal period. In conclusion, the administration of topical EPM formulation at 1 or 1.5 mg/kg may be a valuable tool to be used in regional strategic deworming programs aimed to control ectoparasite infections in dairy production systems.
Assuntos
Lactação , Leite , Administração Tópica , Animais , Bovinos , Feminino , Ivermectina/análogos & derivados , Ivermectina/análise , Leite/químicaRESUMO
Combinations of bioactive phytochemicals with synthetic compounds have been suggested as promissory tools for the improvement of nematode control in livestock. Bioactive phytochemicals may interfere with the activity of drug-metabolizing enzymes and delay the metabolic conversion of anthelmintics into less potent metabolites.This research assessed the effect of the monoterpene thymol (TML) on the in vitro hepatic metabolism of the anthelmintic albendazole (ABZ) in sheep.Liver microsomes from four (4) Texel lambs were incubated with ABZ (50 µM) in absence or in presence of TML (0.05-10 mM).The concentration of TML producing a 50% decrease in ABZ S-oxygenation (IC50) was 13.5 mM. The FMO-dependent S-oxygenation of ABZ was markedly inhibited by the monoterpene (54.1 ± 11.6%, p < .01). In agreement with this observation, TML produced a marked inhibition of benzydamine (BZ) N-oxidase, a specific FMO activity.The CYP-dependent production of the sulfoxide metabolite (ABZSO) was less affected, being 25.3 ± 17.5 lower (p < .05) in presence of TML. Additionally, TML completely abolished the specific CYP1A1-dependent enzyme activity 7-ethoxyresorufin O-deethylase.Overall, the results presented here show that, in addition to its own anthelmintic affect, TML may potentiate ABZ anthelmintic activity by preventing its metabolic conversion into a less active metabolite.
Assuntos
Albendazol/metabolismo , Anti-Helmínticos/metabolismo , Timol/metabolismo , Albendazol/química , Animais , Anti-Helmínticos/química , Citocromo P-450 CYP1A1/metabolismo , Fígado/metabolismo , Taxa de Depuração Metabólica , Microssomos Hepáticos/metabolismo , Monoterpenos , OvinosRESUMO
The combination of the organophosphate (OP) chlorpyrifos (CPF) and the pyrethroid cypermethrin (CPM) is commonly marketed as pour-on formulations for the control of sheep lice, ked, and blowflies. CPF irreversibly inhibits acetylcholinesterases (AChE), while pyrethroids are not AChE inhibitors. However, combinations of pyrethroids with OPs showed a highly synergistic effect on AChE inhibition. Thus, the aim of the current work was to evaluate in vitro and in vivo the inhibitory potency of both pesticides, alone and in combination with AChE and butyrylcholinesterase (BChE) activities in sheep blood. In vitro, IC50 values were similar after CPF or CPF plus CPM incubations. The pour-on coadministration of recommended doses of CPF and CPM did not cause a significant inhibition of AChE and BChE in sheep blood. Only slight percentages of inhibition of their catalytic activities were observed when both drugs were given at 4-fold higher dose rates. The lower systemic availability of topical administration of OPs in sheep may help to explain the lower degree of inhibition of blood AChE and BChE in vivo. The results emerged from this research are a further contribution to the knowledge of the risks of implementing higher dosage regimens of OPs-containing antiparasitic formulations.
Assuntos
Acetilcolinesterase/metabolismo , Butirilcolinesterase/metabolismo , Clorpirifos/efeitos adversos , Inibidores da Colinesterase/efeitos adversos , Piretrinas/efeitos adversos , Ovinos/sangue , Administração Tópica , Animais , Clorpirifos/administração & dosagem , Clorpirifos/uso terapêutico , Inibidores da Colinesterase/administração & dosagem , Inibidores da Colinesterase/uso terapêutico , Combinação de Medicamentos , Inseticidas/administração & dosagem , Inseticidas/efeitos adversos , Inseticidas/uso terapêutico , Masculino , Piretrinas/administração & dosagem , Piretrinas/uso terapêuticoRESUMO
The aim of the current study was to evaluate the in vivo pharmacokinetic of ivermectin (IVM) after the administration of a long-acting (LA) formulation to sheep and its impact on potential drug-drug interactions. The work included the evaluation of the comparative plasma profiles of IVM administered at a single therapeutic dose (200 µg/kg) and as LA formulation at 630 µg/kg. Additionally, IVM was measured in different gastrointestinal tissues at 15 days posttreatment with both IVM formulations. The impact of the long-lasting and enhanced IVM exposure on the disposition kinetics of abamectin (ABM) was also assessed. Plasma (IVM and ABM) and gastrointestinal (IVM) concentrations were analyzed by HPLC with fluorescent detection. In plasma, the calculated Cmax and AUC0-t values of the IVM-LA formulation were 1.47- and 3.35-fold higher compared with IVM 1% formulation, respectively. The T1/2ab and Tmax collected after administration of the LA formulation were 2- and 3.5-fold longer than those observed after administration of IVM 1% formulation, respectively. Significantly higher IVM concentrations were measured in the intestine mucosal tissues and luminal contents with the LA formulation, and in the liver, the increase was 7-fold higher than conventional formulation. There was no drug interaction between IVM and ABM after the single administration of ABM at 15 days post-administration of the IVM LA formulation. The characterization of the kinetic behavior of the LA formulation to sheep and its potential influence on drug-drug interactions is a further contribution to the field.
Assuntos
Anti-Helmínticos/farmacocinética , Ivermectina/farmacocinética , Ovinos/metabolismo , Animais , Anti-Helmínticos/análise , Anti-Helmínticos/sangue , Cromatografia Líquida de Alta Pressão/veterinária , Preparações de Ação Retardada , Interações Medicamentosas , Injeções Subcutâneas , Intestinos/química , Ivermectina/administração & dosagem , Ivermectina/análise , Ivermectina/sangue , Fígado/química , Masculino , Ovinos/parasitologiaRESUMO
The aim of this work was to evaluate the fate of ivermectin (IVM) at two concentrations in cattle feces and its movement to the nearby soil and plants. Feces were spiked with IVM at two levels: 3000 ng g-1 (high group, HG) and 300 ng g-1 (low group, LG). Artificial dung pats were prepared and deposited in an experimental field area. Feces and underlying soil were sampled up to 60 days post-deposition (dpd). As an additional analysis, grasses growing around the pats were sampled at 30 and 60 dpd. Ivermectin concentrations in all matrices were determined by HPLC. Mean IVM fecal concentrations were in the range between 3901.9 ng g-1 and 2419.2 ng g-1 (high group) and 375.3 ng g-1 and 177.49 ng g-1 (low group). Mean times for 50% and 90% dissipation were 88.23 and 293.03 days (HG) and 39.1 and 129.9 days (LG). Soil concentrations ranged from 26.1 ng g-1 to 71.1 ng g-1 (HG) and 3.4 to 5.9 ng g-1 (LG); in plants, concentrations were between 71.4 and 380.8 ng g-1 and 5.40 and 51.8 ng g-1 in HG and LG, respectively. These results confirm that IVM moves from feces to the underlying soil as well as to nearby plants. The potential risk of detrimental effects on soil organisms and the impact on herbivorous animals should be further evaluated.
Assuntos
Fezes/química , Ivermectina/análise , Poluentes do Solo/análise , Animais , Argentina , Bovinos , Cromatografia Líquida de Alta Pressão , Ivermectina/química , Solo/química , Poluentes do Solo/químicaRESUMO
1. Precision-cut liver slices (PCLS) from food-producing animals have not been extensively used to study xenobiotic metabolism, and thus information on this field of research is sparse. 2. The aims of the present work were to further validate the technique of production and culture of bovine PCLS and to characterize the metabolic interaction between the anthelmintic albendazole (ABZ) and the flavin-monooxygenase (FMO) inhibitor methimazole (MTZ). 3. Nine steers were used as donors. PCLS were produced and incubated under two methods: a dynamic organ culture (DOC) incubator and a well-plate (WP) system. 4. Tissue viability, assessed through both structural and functional markers, was preserved throughout 12 h of incubation. ABZ was metabolized to its (+) and (-) albendazole sulfoxide stereoisomers (ABZSO) in bovine PCLS. The interaction between ABZ and MTZ resulted in a reduction (p < 0.001) in the rates of appearance of (+) ABZSO. Conversely, in presence of MTZ, the rates of appearance of (-) ABZSO increased under both systems (p < 0.05). 5. Both culture systems were suitable for assessing the interaction between ABZ and MTZ. 6. Overall, the results presented herein show that PCLS are a useful and reliable tool for short-term studies on metabolic drug-drug interactions in the bovine species.
Assuntos
Interações Medicamentosas , Fígado/metabolismo , Administração Oral , Albendazol/análogos & derivados , Albendazol/metabolismo , Animais , Anti-Helmínticos/metabolismo , Bovinos , Metimazol/metabolismo , Microssomos Hepáticos/metabolismo , EstereoisomerismoRESUMO
In human and mice ATP-binding cassette efflux transporter ABCG2 represents the main route for active drug transport into milk. However, there is no detailed information on the role of ABCG2 in drug secretion and accumulation in milk of dairy animals. We therefore examined ABCG2-mediated drug transport in the bovine mammary gland by parallel pharmacokinetic studies in lactating Jersey cows and in vitro flux studies using the anthelmintic drug monepantel (MNP) as representative bovine ABCG2 (bABCG2) drug substrate. Animals received MNP (Zolvix, Novartis Animal Health Inc.) once (2.5 mg/kg per os) and the concentrations of MNP and the active MNP metabolite MNPSO2 were assessed by high-performance liquid chromatography. Compared with the parent drug MNP, we detected higher MNPSO2 plasma concentrations (expressed as area under the concentration-versus-time curve). Moreover, we observed MNPSO2 excretion into milk of dairy cows with a high milk-to-plasma ratio of 6.75. In mechanistic flux assays, we determined a preferential time-dependent basolateral-to-apical (B > A) MNPSO2 transport across polarized Madin-Darby canine kidney II cells-bABCG2 monolayers using liquid chromatography coupled with tandem mass spectrometry analysis. The B > A MNPSO2 transport was significantly inhibited by the ABCG2 inhibitor fumitremorgin C in bABCG2- but not in mock-transduced MDCKII cells. Additionally, the antibiotic drug enrofloxacin, the benzimidazole anthelmintic oxfendazole and the macrocyclic lactone anthelmintic moxidectin caused a reduction in the MNPSO2(B > A) net efflux. Altogether, this study indicated that therapeutically relevant drugs like the anthelmintic MNP represent substrates of the bovine mammary ABCG2 transporter and may thereby be actively concentrated in dairy milk.
Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Glândulas Mamárias Animais/metabolismo , Leite/metabolismo , Drogas Veterinárias/metabolismo , Animais , Antibacterianos/metabolismo , Transporte Biológico Ativo/fisiologia , Bovinos , Linhagem Celular , Cães , Feminino , Humanos , Lactação/metabolismo , Células Madin Darby de Rim CaninoRESUMO
1. Precision-cut liver slices are one of the in vitro models used in studies concerning xenobiotic metabolism. Sparse information on this field is actually available for cattle and other veterinary species. 2. The aim of the current work was to study the effect of dexamethasone (DEX) on the gene expression and function of CYP3A23 (in rat), CYP3A28 (in cattle) and the transcriptional factors involved in their regulation. 3. DEX (at 100 µM) up-regulated CYP3A23 mRNA (3.2-fold, p = 0.028) in rat liver slices after 12 h culture, whereas the gene expression profiles of transcriptional factors involved in CYP3A regulation were unaffected. A CYP3A-dependent enzyme activity (triacetyl-oleandomycin N-demethylase) increased 3.4-fold (p < 0.05) in rat liver slices cultured in the presence of DEX. 4. The protocol used for rat liver slices was used as reference to study the expression of a CYP3A isoenzyme in cattle liver slices. Oppositely, DEX did neither affect the gene expression profile of CYP3A28 nor the CYP3A activity tested in cattle liver slices. 5. The data reported here are a further contribution to demonstrate the usefulness of liver slices as an in vitro tool for studies on the expression and function of xenobiotic metabolizing enzymes in cattle and in other ruminant species.
Assuntos
Citocromo P-450 CYP3A/genética , Fígado/enzimologia , Animais , Bovinos , Citocromo P-450 CYP3A/metabolismo , Dexametasona/farmacologia , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Isoenzimas/genética , Isoenzimas/metabolismo , L-Lactato Desidrogenase/metabolismo , Fígado/efeitos dos fármacos , Masculino , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/enzimologia , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Sobrevivência de TecidosRESUMO
The activities of different xenobiotic-metabolizing enzymes in liver subcellular fractions from Wistar rats exposed to a glyphosate (GLP)-based herbicide (Roundup full II) were evaluated in this work. Exposure to the herbicide triggered protective mechanisms against oxidative stress (increased glutathione peroxidase activity and total glutathione levels). Liver microsomes from both male and female rats exposed to the herbicide had lower (45%-54%, P < 0.01) hepatic cytochrome P450 (CYP) levels compared to their respective control animals. In female rats, the hepatic 7-ethoxycoumarin O-deethylase (a general CYP-dependent enzyme activity) was 57% higher (P < 0.05) in herbicide-exposed compared to control animals. Conversely, this enzyme activity was 58% lower (P < 0.05) in male rats receiving the herbicide. Lower (P < 0.05) 7-ethoxyresorufin O-deethlyase (EROD, CYP1A1/2 dependent) and oleandomycin triacetate (TAO) N-demethylase (CYP3A dependent) enzyme activities were observed in liver microsomes from exposed male rats. Conversely, in females receiving the herbicide, EROD increased (123%-168%, P < 0.05), whereas TAO N-demethylase did not change. A higher (158%-179%, P < 0.01) benzyloxyresorufin O-debenzylase (a CYP2B-dependent enzyme activity) activity was only observed in herbicide-exposed female rats. In herbicide-exposed rats, the hepatic S-oxidation of methimazole (flavin monooxygenase dependent) was 49% to 62% lower (P < 0.001), whereas the carbonyl reduction of menadione (a cytosolic carbonyl reductase-dependent activity) was higher (P < 0.05). Exposure to the herbicide had no effects on enzymatic activities dependent on carboxylesterases, glutathione transferases, and uridinediphospho-glucuronosyltransferases. This research demonstrated certain biochemical modifications after exposure to a GLP-based herbicide. Such modifications may affect the metabolic fate of different endobiotic and xenobiotic substances. The pharmacotoxicological significance of these findings remains to be clarified.
Assuntos
Glicina/análogos & derivados , Herbicidas/toxicidade , Fígado/efeitos dos fármacos , Intoxicação por Organofosfatos/enzimologia , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Xenobióticos/metabolismo , O-Dealquilase 7-Alcoxicumarina/antagonistas & inibidores , O-Dealquilase 7-Alcoxicumarina/química , O-Dealquilase 7-Alcoxicumarina/metabolismo , Animais , Carbonil Redutase (NADPH)/química , Carbonil Redutase (NADPH)/metabolismo , Citocromo P-450 CYP1A1/antagonistas & inibidores , Citocromo P-450 CYP1A1/química , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A2/química , Citocromo P-450 CYP1A2/metabolismo , Citocromo P-450 CYP2B1/química , Citocromo P-450 CYP2B1/metabolismo , Citocromo P-450 CYP3A/química , Citocromo P-450 CYP3A/metabolismo , Relação Dose-Resposta a Droga , Feminino , Glicina/administração & dosagem , Glicina/toxicidade , Herbicidas/administração & dosagem , Fígado/enzimologia , Fígado/metabolismo , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Microssomos Hepáticos/metabolismo , Intoxicação por Organofosfatos/metabolismo , Oxigenases/antagonistas & inibidores , Oxigenases/metabolismo , Distribuição Aleatória , Ratos Wistar , Caracteres Sexuais , Poluentes Químicos da Água/administração & dosagem , GlifosatoRESUMO
The anthelmintic fenbendazole (FBZ) undergoes hepatic Soxygenation by monooxygenases belonging to the cytochrome P450 (CYP) and flavin-monooxygenase (FMO) families. The in-feed medication with FBZ induced CYP1A-dependent metabolism in pig liver. This fact may alter the metabolism of the anthelmintic itself, and of CYP1A substrates like aflatoxin B1 (AFB1). This work evaluated the effect of the in-feed administration of FBZ on CYP1A-dependent metabolism, on its own pattern of hepatic Soxygenation, and on the metabolism of AFB1. Landrace piglets remained untreated (n = 5) or received a pre-mix of FBZ (n = 6) in feed for 9 days. Pigs were slaughtered for preparation of liver microsomes used for: CYP content determination; monitoring the CYP1A-dependent enzyme activities, 7-ethoxyresorufin O-deethylase (EROD) and 7-methoxyresorufin O-demethylase (MROD); measurement of FBZ (50 µM) Soxygenation, and AFB1 (16 nM) disappearance from the incubation medium. In microsomes of FBZ-treated animals, EROD and MROD increased 19-fold (p = 0.002) and 14-fold (p = 0.003), respectively. An enhanced (3-fold, p = 0.004) participation of the CYP pathway in FBZ Soxygenation was observed in the liver of piglets treated with the anthelmintic (210 ± 69 pmol/min.nmol CYP) compared to untreated animals (68 ± 34 pmol/min.nmol CYP). AFB1 metabolism was 93% higher (p = 0.009) in the liver of FBZ-treated compared to untreated pigs. Positive and significant (p < 0.05) correlations were observed between CYP1A-dependent enzyme activities and FBZ or AFB1 metabolism. The sustained administration of FBZ caused an auto-induction of the CYP1A-dependent Soxygenation of this anthelmintic. The CYP1A induction triggered by the anthelmintic could amplify the production of AFB1 metabolites in pig liver, including the hepatotoxic AFB1-derived epoxide.+.
Assuntos
Anti-Helmínticos , Citocromo P-450 CYP1A1 , Humanos , Animais , Suínos , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A1/farmacologia , Fenbendazol/farmacologia , Fenbendazol/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Anti-Helmínticos/farmacologia , Microssomos Hepáticos/metabolismo , Interações MedicamentosasRESUMO
Precision-cut liver slices (PCLS) are tissue explants extensively used as an ex vivo model for metabolism and toxicity studies. When in vitro assays are conducted, it is imperative to perform a histomorphological evaluation as part of the viability analyses throughout the assay time. It is considered that good quality PCLS histological sections are difficult to obtain because they are hard to manipulate, and may shrink or fold during processing. Moreover, bibliography is not detailed on the embedding processes used. In this article, we propose an adjusted and rapid method for paraffin embedding of PCLS from crossbreed steers. Each PCLS was covered with a piece of gauze and placed into a histological cassette. These cassettes were submitted to a series of baths: 80â¯% ethanol for 10â¯min; 3 baths of 96â¯% ethanol for 10â¯min; 3 baths of butanol for 10â¯min; 1 bath of butanol-paraffin (1:1) for 20â¯min in a 60 °C laboratory oven; and 3 baths of paraffin for 20â¯min in a 60 °C laboratory oven. Folded paper boxes were used to produce paraffin blocks. It was possible to obtain complete sections with preserved cell morphology and no artifacts, and tissue appearance was similar to previous PCLS processed through the routine protocol, demonstrating the adequacy of the method implemented.
RESUMO
This review outlines the current state of anthelmintic resistance (AHR) of gastrointestinal nematodes (GINs) among cattle in Argentina and Brazil, emphasizing the economic repercussions, animal health and welfare. The analysis explores factors associated with AHR and proposes a potential solution: the use of drug combinations. Both countries are grappling with a severe AHR scenario in cattle, having progressed through incipient, established, and advanced phases, leading to extreme cases of animal mortality due to ineffective control strategies. Genera such as Cooperia and Haemonchus have the highest reports of resistance, with Oesophagostomum radiatum also posing significant problems. While oral benzimidazoles and levamisole remain effective in most herds, moxidectin is entering an advanced resistance phase, and avermectins are increasingly deemed ineffective. The review explores the impact ofclimate, mixed grazing, animal movement and other husbandry practices, and the relationship between ectoparasite control and the emergence of resistant helminths. Notably, the discussion includes the strategic use of drug combinations as a valuable approach to address resistant GINs control in livestock, highlighting its significant potential to mitigate the challenges posed by AHR in the cattle industry of these countries.
Assuntos
Anti-Helmínticos , Doenças dos Bovinos , Resistência a Medicamentos , Nematoides , Infecções por Nematoides , Animais , Bovinos/parasitologia , Brasil , Argentina , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/parasitologia , Infecções por Nematoides/veterinária , Infecções por Nematoides/tratamento farmacológico , Infecções por Nematoides/parasitologia , Anti-Helmínticos/uso terapêutico , Nematoides/efeitos dos fármacosRESUMO
Fenbendazole (FBZ), a benzymidazole (BZD) anthelmintic drug, is used for in-feed medication in pigs. BZD-containing drugs may induce cytochrome P450 isozymes (CYPs), particularly those members of the CYP1A subfamily. The current research evaluated the plasma and liver availability and metabolism of FBZ and its metabolites, oxfendazole (OFZ) and fenbendazole sulphone (FBZSO2), after the administration of the parent drug in feed, and characterized the effect of the sustained administration of the anthelmintic on the catalytic activities of xenobiotic metabolizing enzymes in pig liver. Five female Landrace piglets remained untreated (controls), and other six were treated with a pre-mix of FBZ, combined with feed, for 9 consecutive days as usually is recommended. Blood samples were collected from each treated animal up to day 9 and analyzed by HPLC; all animals were slaughtered for preparation of liver microsomes. Plasma concentration ratios OFZ/FBZ and FBZSO2/OFZ increased significantly (p < 0.05) from the beginning to the end of drug exposure, which may indicate an enhanced conversion of FBZ into its metabolites. FBZ represented 45.8 ± 3.4% of the total anthelmintic molecules in liver tissue. Increased CYP1A-dependent 7-ethoxy (24.5-fold, p = 0.0032) and 7-methoxyresorufin (17.2-fold, p = 0.0006) O-dealkylase activities was observed in liver microsomes from FBZ-treated animals. In addition, a 64% increase (p = 0.042) in the rate of FBZ S-oxidation was observed in pigs treated with the anthelmintic drug compared to that measured in untreated animals. Thus, the continuous FBZ administration may accelerate its own in vivo hepatic metabolism through the CYP1A pathway.
Assuntos
Anti-Helmínticos , Fenbendazol , Animais , Feminino , Suínos , Fenbendazol/farmacologia , Fenbendazol/metabolismo , Xenobióticos/metabolismo , Anti-Helmínticos/farmacologia , Anti-Helmínticos/metabolismo , Fígado/metabolismoRESUMO
Psoroptic mange causes relevant losses of productivity in cattle. Macrocyclic lactones are one of the main pharmacological tools recommended for controlling it. The aim of the current work was to compare the relationship between the pharmacokinetic behavior and the effectiveness of both ivermectin (IVM) and doramectin (DRM) following their administration as either the traditional (1 %) or long-acting (3.15-3.5 %) injectable formulations to cattle naturally infected with Psoroptes ovis. The overall work involved three trials (1, 2 and 3) carried out on commercial beef cattle farms (grazing systems). In Trial 1, 20 grazing steers with active mange infection were allocated into 2 groups (n = 10) and treated subcutaneously (SC) with either IVM (1 %) or DRM (1%) at 0.2 mg/kg. In Trial 2, 16 grazing steers with active mange divided in 2 groups (n = 8) were treated SC with either IVM 1 % (0.2 mg/kg) or IVM 3.15 % long-acting (0.63 mg/kg). In Trial 3, 2 groups of mange infected steers (n = 8) were treated SC with either IVM 3.15 % (0.63 mg/kg) or DRM 3.5 % (0.7 mg/kg). Blood samples were collected of each experimental group and the drug systemic availability was estimated by measuring of IVM/DRM concentrations by HPLC. Skin scraping samples were collected from each animal and mites were counted at 14, 21 and 28 days post-treatment. In Trial 1, the mite density score on day 14 was significantly lower for DRM (0.60) compared to IVM (1.80) (P = 0.019). Based on the number of animals clinically cured (negative to the presence of mites), the efficacy of DRM was higher (80 %) than that obtained for IVM (10 %) (P < 0.05). DRM systemic exposure measured as AUC was 1.37-fold higher compared to IVM. In Trial 2, even though IVM exposure was significantly greater after the long-acting (3.15 %) compared to the traditional formulation (1 %), none of the treatments significantly reduced the mite density score, with a percentage of animals cured between 0 % and 37.5 % after both IVM treatments. In Trial 3, the 100 % of cured animals were achieved at day 21 (IVM 3.15 %) and at day 28 (DRM 3.5 %) post-treatment. In conclusion, DRM treatment could offer some therapeutic advantages in field situations where IVM fails to control mange. Depending on the level of susceptibility of the mite population, long-acting pharmaceutical formulations can be useful to control Psoroptic mange in cattle. The use of macrocyclic lactones for mange control in cattle should be based on appropriate diagnosis on each individual farm.
Assuntos
Doenças dos Bovinos , Infestações por Ácaros , Ácaros , Animais , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/prevenção & controle , Ivermectina/farmacocinética , Infestações por Ácaros/tratamento farmacológico , Infestações por Ácaros/prevenção & controle , Infestações por Ácaros/veterinária , Distribuição AleatóriaRESUMO
Ivermectin (IVM), one of the most widely used antiparasitics in livestock, could enters into the aquatic environment because the treated animal metabolizes only a small percentage of what is administered and the rest is eliminated through the feces, largely as a parent drug, imposing a risk to aquatic organisms. The aims of this study were to (1) assess the effect of IVM spiked in cattle dung on the survival and emergence of Culex pipiens (Diptera: Culicidae), and to (2) evaluate the accumulation of this drug in the different developmental stages of this taxon. Larvae were exposed to two IVM concentrations (T1: 1000 ng g-1 and T2: 500 ng g-1) for 9 days. At days 3, 6 and 9 survival and adult emergence were recorded and samples of larvae, pupae, pupal exuviae and adults were taken to analyze the IVM accumulation. At these concentrations, a reduction in survival and adult emergence of C. pipiens was recorded. In addition, the IVM accumulation was observed in all samples analyzed, decreasing it throughout the development of this taxon (larvae > pupae > adults). Although a large proportion of the drug was lost during the metamorphosis, being mainly eliminated through pupal exuviae during molting, this process is not enough to eliminate it completely. Thus, part of the drug was transferred to the adult stage and remains available to the aquatic and terrestrial food webs. These results show that IVM represents a risk to aquatic invertebrates and their predators, which deserves further studies, especially in the context of their bioaccumulation and biomagnification through the aquatic and terrestrial trophic webs.