Embrapa Pecuaria Sudeste CPPSE

São Carlos, Brazil

Embrapa Pecuaria Sudeste CPPSE

São Carlos, Brazil
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Chagas A.C.S.,Embrapa Pecuaria Sudeste CPPSE | Domingues L.F.,Embrapa Pecuaria Sudeste CPPSE | Barioni-Junior W.,Embrapa Pecuaria Sudeste CPPSE | Esteves S.N.,Embrapa Pecuaria Sudeste CPPSE | Niciura S.C.M.,Embrapa Pecuaria Sudeste CPPSE
Parasitology Research | Year: 2016

Levamisole phosphate, chosen based on its 100 % efficacy demonstrated by a previous fecal egg count reduction test (FECRT), was used as the exclusive anthelmintic treatment in the Embrapa Southeast Livestock sheep flock from 2009 to 2014 in a target selected treatment scheme. In the present study, the effectiveness of this nematode control scheme was evaluated after 5 years by FECRT, larval development test (LDT), and a molecular test to assess the development of levamisole resistance in Haemonchus contortus. Animals were submitted to treatments with albendazole, levamisole, closantel, ivermectin, moxidectin, and monepantel. Eggs per gram of feces (EPG) counts and fecal cultures were performed, and anthelmintic efficacy was calculated by the RESO 4.0 program. The helminths of the flock (GIN Embrapa2014) were compared to susceptible (McMaster) and resistant (Embrapa2010) H. contortus isolates in the LDT to estimate the LC50 and LC90 of levamisole and in a molecular test to evaluate the 63-bp indel in the acr8 gene associated with levamisole resistance. In the FECRT, parasites were susceptible to monepantel (99.6 %) and closantel (98.3 %), but resistant to moxidectin (93.8 %), levamisole (70.4 %), ivermectin (48.1 %), and albendazole (0 %). In the coproculture on D14, and the control group presented 80 % H. contortus and 20 % Trichostrongylus sp., while in the monepantel group L1 were observed as well as Oesophagostomum sp. L3. LDT and resistance factors provided good separation between susceptible and resistant parasites. The genotypic frequencies of the 63-bp insertion in the acr8 gene in H. contortus were 11.9, 6.7, and 0 % in GIN Embrapa2014, Embrapa2010, and McMaster isolates, respectively. After 5 years of exclusive use, the nematodes developed resistance to levamisole, detected by FECRT and by increase in LC50 and LC90 for levamisole in the LDT. The 63-bp indel was not confirmed as a molecular marker of levamisole resistance in our isolates. The target selected treatment scheme was effective to control helminths in the sheep flock for 5 years, when levamisole’s inefficacy was perceived because of no change in the clinical situation of treated animals. Through this scheme, it was possible to promote reversion towards susceptibility or increase of efficacy for other chemical classes. Thus, this is a valid recommendation to control worms and to delay the development of resistance, preserving other anthelmintic classes for future use. © 2015, Springer-Verlag Berlin Heidelberg.


Domingues L.F.,Embrapa Pecuaria Sudeste CPPSE | Rabelo M.D.,Embrapa Pecuaria Sudeste CPPSE | Lopez E.R.,Agricultural University of Havana | Chagas A.C.S.,Embrapa Pecuaria Sudeste CPPSE
Industrial Crops and Products | Year: 2015

Gastrointestinal nematodes are an important problem in the small ruminant production and their control nearly exclusively depends on commercial anthelmintics. However, parasite resistance to those chemical formulations is an increasing global problem, so new plant-derived compounds are being studied for their potential use against gastrointestinal nematodes. Citrus sinensis and Melaleuca quinquenervia essential oils were evaluated against Haemonchus contortus Embrapa2010 resistant isolate, through the egg hatch test (EHT) and larval development test (LDT) at concentrations ranging from 0.02mg/mL to 50mg/mL and from 0.04mg/mL to 3.12mg/mL, respectively. All concentrations, positive controls (thiabendazole in EHT or ivermectin in LDT), and negative controls (2% Tween 80 in EHT or 0.5% DMSO in LDT) were performed in six replicates and in three independent experiments, for a total of 18 repetitions involving approximately 1800 parasites per treatment. Significant differences (P≤0.01) of inhibition percentages were identified by one-way analysis of variance followed by the Tukey test. Inhibitory concentration values (IC50 and IC90) were analyzed by the Probit procedure of SAS. The essential oils evaluated showed that C. sinensis presented limonene as major component (96.0%), while M. quinquenervia contained longifolene (32.95%) and 1,8-cineole (25.43%) as major components. In the EHT, the IC50 and IC90 of the essential oils were respectively 0.27 and 0.99mg/mL for C. sinensis and, 1.52 and 5.63mg/mL for M. quinquenervia. In the LDT, the IC50 and IC90 were 0.97 and 2.32mg/mL for C. sinensis and, 0.44 and 0.94mg/mL for M. quinquenervia. C. sinensis was five times more effective on eggs than M. quinquenervia. However, it was twice more effective on larvae than C. sinensis, indicating that bioactive essential oils have different modes of action. The results suggest that these compounds are good candidates for nematode control. However, alternative anthelmintics have to be safe to the host, promote parasite control in vivo and not generate residues. So, all these key points need to be elucidated before using any plant extract and/or components. © 2015.


PubMed | Embrapa Pecuaria Sudeste CPPSE
Type: Journal Article | Journal: Parasitology research | Year: 2016

Levamisole phosphate, chosen based on its 100 % efficacy demonstrated by a previous fecal egg count reduction test (FECRT), was used as the exclusive anthelmintic treatment in the Embrapa Southeast Livestock sheep flock from 2009 to 2014 in a target selected treatment scheme. In the present study, the effectiveness of this nematode control scheme was evaluated after 5 years by FECRT, larval development test (LDT), and a molecular test to assess the development of levamisole resistance in Haemonchus contortus. Animals were submitted to treatments with albendazole, levamisole, closantel, ivermectin, moxidectin, and monepantel. Eggs per gram of feces (EPG) counts and fecal cultures were performed, and anthelmintic efficacy was calculated by the RESO 4.0 program. The helminths of the flock (GIN Embrapa2014) were compared to susceptible (McMaster) and resistant (Embrapa2010) H. contortus isolates in the LDT to estimate the LC50 and LC90 of levamisole and in a molecular test to evaluate the 63-bp indel in the acr8 gene associated with levamisole resistance. In the FECRT, parasites were susceptible to monepantel (99.6 %) and closantel (98.3 %), but resistant to moxidectin (93.8 %), levamisole (70.4 %), ivermectin (48.1 %), and albendazole (0 %). In the coproculture on D14, and the control group presented 80 % H. contortus and 20 % Trichostrongylus sp., while in the monepantel group L1 were observed as well as Oesophagostomum sp. L3. LDT and resistance factors provided good separation between susceptible and resistant parasites. The genotypic frequencies of the 63-bp insertion in the acr8 gene in H. contortus were 11.9, 6.7, and 0 % in GIN Embrapa2014, Embrapa2010, and McMaster isolates, respectively. After 5 years of exclusive use, the nematodes developed resistance to levamisole, detected by FECRT and by increase in LC50 and LC90 for levamisole in the LDT. The 63-bp indel was not confirmed as a molecular marker of levamisole resistance in our isolates. The target selected treatment scheme was effective to control helminths in the sheep flock for 5 years, when levamisoles inefficacy was perceived because of no change in the clinical situation of treated animals. Through this scheme, it was possible to promote reversion towards susceptibility or increase of efficacy for other chemical classes. Thus, this is a valid recommendation to control worms and to delay the development of resistance, preserving other anthelmintic classes for future use.


Torres T.R.,University of Pernambuco | Do C.m.m. Ludke M.,University of Pernambuco | Do C.m.m. Ludke M.,Brazilian National Council for Scientific and Technological Development | MacIel M.I.S.,University of Pernambuco | And 4 more authors.
Revista Brasileirade Ciencias Agrarias | Year: 2011

The sensory attributes of the meat from broilers fed from the seventh to the 42nd day of age with diets containing different replacement levels of the soybean meal protein by the extruded cottonseed meal protein (FAE) were evaluated by the Simplified Quantitative Descriptive Analysis and the Triangle Test. Five treatments (0, 10, 20, 30 and 40 % replacement, respectively, T1, T2, T3, T4 and T5) and five replications per treatment containing 12 birds per plot were used. At the 42nd day of age, three broilers per plot, weighing close to the plot average, were slaughtered, and the 75 carcasses were frozen. In the sensorial evaluation, 15 g portions of the breast and thigh+drumstick cuts from the carcasses obtained in T1, T2, T3 and T5 were removed, cut into cubes of 2.5 cm2 and cooked. The samples were evaluated by a team of ten trained tasters. The analyzed attributes showed no significant difference (P>0.05) between treatments. However, by the difference test (Triangle Test), significant difference was observed (P<0.05) in the attribute thigh+drumstick meat taste when the treatments of lower (T2) and higher FAE inclusion level (T5) were compared to the treatment without FAE (T1).


de Souza Chagas A.C.,Embrapa Pecuaria Sudeste CPPSE | de Sena Oliveira M.C.,Embrapa Pecuaria Sudeste CPPSE | Giglioti R.,Embrapa Pecuaria Sudeste CPPSE | Santana R.C.M.,Embrapa Pecuaria Sudeste CPPSE | And 3 more authors.
Ticks and Tick-borne Diseases | Year: 2016

Herbal extracts have been investigated as an alternative for parasite control, aiming to slow the development of resistance and to obtain low-cost biodegradable parasiticides. The goal of this study was to evaluate the efficacy, in vitro, of 11 essential oils from Brazil on reproductive efficiency and lethality of the cattle tick Rhipicephalus (Boophilus) microplus. The effects of oils extracted from Curcuma longa, Zingiber officinale, Lippia alba, Lippia gracilis, Lippia origanoides, Lippia sidoides, Mentha arvensis, Mentha piperita, Croton cajucara (white and red), and Croton sacaquinha on ticks were investigated by the Immersion Test with Engorged Females (ITEF) and the modified Larval Packet Test (LPT). Distilled water and 2% Tween 80 were used as control treatments. Chemical analysis of the oils was done with gas chromatography coupled with mass spectrometry. Analysis of the in vitro tests using Probit (SAS program) allowed the calculation of lethal concentrations (LCs). Lower reproductive efficiency indexes and higher efficacy percentages in the ITEF were obtained with the oils extracted from C. longa (24 and 71%, respectively) and M. arvensis oils (27 and 73%, respectively). Lower LC50 was reached with C. longa (10.24 mg/mL), L. alba (10.78 mg/mL), M. arvensis (22.31 mg/mL), L. sidoides (27.67 mg/mL), and C. sacaquinha (29.88 mg/mL) oils. In the LPT, species from Zingiberaceae and Verbenaceae families caused 100% lethality at 25 mg/mL, except for L. sidoides. The most effective oils were from C. longa, L. gracilis, L. origanoides, L. alba, and Z. officinale. The LC50 and LC90 were, respectively: 0.54 and 1.80 mg/mL, 3.21 and 7.03 mg/mL, 3.10 and 8.44 mg/mL, 5.85 and 11.14 mg/mL, and 7.75 and 13.62 mg/mL. The efficacy was directly related to the major components in each essential oil, and the oils derived from Croton genus presented the worst performance, suggesting the absence of synergistic effect among its compounds. Since C. longa, containing 62% turmerone, was the one most efficient against ticks, this compound may be potentially used for tick control, but further research is needed, especially to assess toxicity of these compounds to the host. These new studies, together with the results presented here, may provide a strong rationale for designing pre-clinical and clinical studies with these agents. © 2016 Elsevier GmbH.


Moreira A.,Embrapa Pecuaria Sudeste CPPSE | de Castro C.,Embrapa Soja CNPSO | de Oliveira F.A.,Embrapa Soja CNPSO
Ciencia e Agrotecnologia | Year: 2010

The objective of this work was to evaluate the efficiency of six extractants and the effect of boron concentration and dry matter yield, cultivated in greenhouse conditions with two Rhodic Hapludox in Mauá da Serra and Londrina, in the State of Paraná, and one Ustoxix Quatzipsamment in Alto Garças, State of Mato Grosso, Brazil. The experiment was conducted in a completely randomized design with six boron rates (0, 0.25, 0.5, 1, 2 and 4 kg ha-1), and three replicates. The available boron was determined with six extractant solutions: Mehlich 1, Mehlich 3, hot water, HCl 0.05 mol L-1, Ca(H2PO4)2.H2O 0.01 mol L-1 and KCl 1.0 mol L-1. The results showed that the B extracted by hot water had the best correlation with the dry matter yield and B content in the plant, whereas the KCl 1.0 mol L-1 was the most efficient in the determination of B level in dry matter. High concentrations of B diminished the efficiency of the hot water extractant in determining the B available in soil. The highest correlations were obtained between the extractant Mehlich 1 and Ca(H2PO4)2.H2O 0.01 mol L-1, KCl 1.0 mol L-1 and HCl 0.05 mol L-1, KCl 1.0 mol L-1 and Ca(H2PO4)2.H2O 0.01 mol L-1, and hot water and Ca(H2PO4)2.H2O 0.01 mol L-1.


Melo P.C.,São Paulo State University | Ferreira L.M.,Medico Veterinario Autonomo | Nader-Filho A.,São Paulo State University | Zafalon L.F.,Embrapa Pecuaria Sudeste CPPSE | Vicente H.I.G.,Secretaria da Agricultura do Estado de Sao Paulo
Bioscience Journal | Year: 2012

Biofilm formation is considered an advantage for Staphylococcus aureus mastitis isolates, facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers and enclosing in a polymeric matrix known as exopolysaccharide. The objective of this study was to evaluate the ability of Staphylococcus aureus isolated from bovine subclinical mastitis for formation of biofilms. A total of 94 Staphylococcus aureus strains obtained from milk samples of cows suffering from subclinical mastitis in dairy herds on two properties in the state of São Paulo were evaluated. These strains were characterized by in vitro biofilm formation, and by the presence of icaA and icaD genes which are responsible for intercellular adhesion. The results revealed that 98.9% of the isolates produced biofilm in vitro by adherence in sterile 96-well "U" bottom polystyrene tissue culture plates; 95.7% of the isolates possessed the icaA and icaD genes. These bacterial isolates biofilm producers may impair eradication of chronic mastitis, rendering antibiotherapy less effective. The detection of biofilm forming ability in mastitis isolates may provide useful information for more adequate therapeutic regimen and for preventive actions in the control of those bacterial isolates in bovine herds.


PubMed | Embrapa Pecuaria Sudeste CPPSE and University of Ribeirão Preto
Type: | Journal: Veterinary parasitology | Year: 2016

Haemonchus contortus is an important gastrointestinal parasite on sheep farms in tropical regions. The resistance of the parasite against most anthelmintic drugs represents a great economic problem to sheep farming and is a major challenge that needs to be overcome. The searches for new anthelmintic agents that act on different stages of the parasites life cycle are necessary for the development of new therapeutic options. The aim of this study was to evaluate the in vitro and in vivo anthelmintic activity of Thymus vulgaris essential oil against H. contortus and of its main component, the monoterpene thymol. Despite the relative ineffectiveness of the oil in the in vivo test, which may be corrected in the future after technical improvements to increase the oils bioavailability, the in vitro results validated the popular use of T. vulgaris oil as an anthelmintic agent, at least against H. contortus. In fact, both the essential oil and thymol, which accounts for 50.22% of the oil composition, were effective against the three main stages of H. contortus. The oil and thymol were able to inhibit egg hatching by 96.4-100%, larval development by 90.8-100%, and larval motility by 97-100%. Similar to the positive control (levamisole 20mg/mL), the oil and thymol completely inhibited the motility of H. contortus adults within the first 8h of the experiment. Since thymol reproduces the anthelmintic effects of the oil and because it is the main component of the oil, it is reasonable to assume that thymol is the most important compound responsible for the anthelmintic effect of T. vulgaris. These results are of ethnopharmacological importance and may contribute to the development of new drugs and even herbal medicines, increasing treatment options for the farm breeding.


PubMed | Embrapa Amazonia Ocidental, Embrapa Pecuaria Sudeste CPPSE and Embrapa Agroindustria de Alimentos
Type: Journal Article | Journal: Ticks and tick-borne diseases | Year: 2016

Herbal extracts have been investigated as an alternative for parasite control, aiming to slow the development of resistance and to obtain low-cost biodegradable parasiticides. The goal of this study was to evaluate the efficacy, in vitro, of 11 essential oils from Brazil on reproductive efficiency and lethality of the cattle tick Rhipicephalus (Boophilus) microplus. The effects of oils extracted from Curcuma longa, Zingiber officinale, Lippia alba, Lippia gracilis, Lippia origanoides, Lippia sidoides, Mentha arvensis, Mentha piperita, Croton cajucara (white and red), and Croton sacaquinha on ticks were investigated by the Immersion Test with Engorged Females (ITEF) and the modified Larval Packet Test (LPT). Distilled water and 2% Tween 80 were used as control treatments. Chemical analysis of the oils was done with gas chromatography coupled with mass spectrometry. Analysis of the in vitro tests using Probit (SAS program) allowed the calculation of lethal concentrations (LCs). Lower reproductive efficiency indexes and higher efficacy percentages in the ITEF were obtained with the oils extracted from C. longa (24 and 71%, respectively) and M. arvensis oils (27 and 73%, respectively). Lower LC50 was reached with C. longa (10.24 mg/mL), L. alba (10.78 mg/mL), M. arvensis (22.31 mg/mL), L. sidoides (27.67 mg/mL), and C. sacaquinha (29.88 mg/mL) oils. In the LPT, species from Zingiberaceae and Verbenaceae families caused 100% lethality at 25 mg/mL, except for L. sidoides. The most effective oils were from C. longa, L. gracilis, L. origanoides, L. alba, and Z. officinale. The LC50 and LC90 were, respectively: 0.54 and 1.80 mg/mL, 3.21 and 7.03 mg/mL, 3.10 and 8.44 mg/mL, 5.85 and 11.14 mg/mL, and 7.75 and 13.62 mg/mL. The efficacy was directly related to the major components in each essential oil, and the oils derived from Croton genus presented the worst performance, suggesting the absence of synergistic effect among its compounds. Since C. longa, containing 62% turmerone, was the one most efficient against ticks, this compound may be potentially used for tick control, but further research is needed, especially to assess toxicity of these compounds to the host. These new studies, together with the results presented here, may provide a strong rationale for designing pre-clinical and clinical studies with these agents.

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