Effect of the Combination of Synthetic Anthelmintics with Carvacryl Acetate in Emulsions with and without a Sodium Alginate Matrix on Haemonchus contortus.

The present study aimed to evaluate the effect of nanoemulsions using combined synthetic anthelmintics, thiabendazole (TBZ), levamisole (LEV), and ivermectin (IVM), with carvacryl acetate (CA) against Haemonchus contortus, and also tested the presence and absence of alginate (ALG). The anthelmintic effect of the CA/TBZ nanoemulsion was evaluated in the egg hatch test (EHT). The effects of CA/IVM and CA/LEV nanoemulsions were evaluated in the larval development test (LDT). The emulsions CA/TBZ/ALG and CA/TBZ showed a multimodal profile, with most particles on the nanometric scale. The encapsulation efficiency in CA/TBZ/ALG was 80.25%, and that in CA/LEV/ALG was 89.73%. In the EHT, CA/TBZ and CA/TBZ/ALG showed mean combination indices (CIs) of 0.55 and 0.36, respectively, demonstrating synergism in both. In LDT, CA/IVM had an average CI of 0.75, and CA/LEV and CA/LEV/ALG showed CI values of 0.4 and 0.93, respectively. It was concluded that CA/TBZ showed a synergistic interaction, and CA/TBZ/ALG showed an enhanced effect. In addition, the matrix brought stability to the product, encouraging its improvement to obtain higher efficacy.

Anthelmintic resistance in sheep in the semiarid region of Minas Gerais, Brazil.

Anthelmintic efficacy was evaluated among sheep that had become naturally infected with gastrointestinal nematodes in 17 flocks located in the semiarid region of Minas Gerais, Brazil. Feces were collected individually from 1021 hairy sheep to determine the number of eggs per gram of feces (EPG) and for coprocultures to identify nematode genera the nematodes. Only the animals that presented EPG counts greater than or equal to 200 were included in the study (totaling 381 sheep). The animals were divided into three treatment groups: albendazole, ivermectin and levamisole. Fourteen days after the administration of anthelmintics, fecal samples were taken from all animals. In each flock, the pre-treatment and post-treatment arithmetic mean EPG were used to calculate the efficacy (FECR) for each of the treatment groups and the lower 95% confidence limit. Data were analyzed with the "eggCounts 2.3" package in RStudio, using a Bayesian model for paired design. The anthelmintics were classified as being efficacious (when the FECR was both equal to or above 95% and the lower 95% confidence limit was equal to or above 90%) or as encountering anthelmintic resistance (when the FECR was below 95% and the lower 95% confidence limit was below 90%) or inconclusive (when none of the other criteria were fulfilled). Albendazole and ivermectin were not effective in any of the flocks. Levamisole was effective against gastrointestinal nematodes in 25% of the flocks studied. Haemonchus, Trichostrongylus and Oesophagostomum genera were identified in this study in a semiarid region of Minas Gerais, Brazil. The genus Haemonchus was the most prevalent, followed by Trichostrongylus and Oesophagostomum. After anthelmintic treatment, the most prevalent genus was Haemonchus, followed by Trichostrongylus; the genus Oesophagostomum was not detected. The highest percentage of Haemonchus larvae was observed after treatment with ivermectin, followed by albendazole and levamisole. This study revealed the existence of gastrointestinal nematodes in sheep that present multiple resistance to all three main classes of anthelmintic drugs.

Anthelmintic resistance against gastrointestinal nematodes in beef cattle: effect on liveweight gain of grazing livestock in Mexico / Resistência anti-helmíntica em nematódeos gastrointestinais de bovinos de corte em uma região tropical: efeito no ganho de peso vivo do gado de pasto no México

This study aimed to determine the efficacy and parasite resistance of levamisole (LV) and ivermectin (IVM) in beef cattle naturally infected with gastrointestinal nematodes, as well as the effect on the liveweight gain in a tropical wet region of Oaxaca, Mexico. From November 2019 to January 2020, sixty-six grazing calves were randomly allocated into three groups of twenty-two animals each, treated with LV or IVM or an untreated control group (day 0). Feces were collected 1 day before treatment and 15 days after treatment. The liveweight gain from each animal was recorded at days 0, 15, 30 and 45 post treatment. The LV group presented the highest reduction of eggs per gram (EPG) of feces, followed by the IVM group. Resistance to IVM was detected, although LV resistance was also suspected. The IVM group had significantly higher effective treatment at 93.5%, resulting in an increase (P<0.05) of liveweight gain of 16.1kg, followed by the LV group (92.4%) with 17.1kg, compared to the untreated control group. A significant (P < 0.05) negative correlation was observed between EPG and weight gain for the LV (r = -0.46) and IVM groups (r = -0.32). LV and IVM showed a lack of efficacy against gastrointestinal nematodes, as well as an adequate capacity for EPG reduction but with IVM resistance and detrimental effects on growth performance in grazing beef cattle.

Os nematódeos gastrointestinais do gado de pastoreio causam perdas econômicas substanciais em todo o mundo. Este estudo teve como objetivo determinar a eficácia e a resistência parasitária do levamisol (LV) e da ivermectina (IVM) em bovinos de corte naturalmente infectados com nematódeos gastrointestinais, bem como o efeito no ganho de peso vivo, em uma região tropical úmida de Oaxaca, México. De novembro de 2019 a janeiro de 2020, 66 bezerros de pasto foram distribuídos aleatoriamente em três grupos de 22 animais cada um, tratados com LV ou IVM, ou em um grupo controle sem tratamento (dia 0). As fezes foram coletadas 1 dia antes do tratamento e 15 dias após o tratamento. O ganho de peso vivo de cada animal foi registrado nos dias 0, 15, 30 e 45 pós-tratamento. O grupo do LV apresentou a maior redução de ovos por grama de fezes (EPG), seguido pelo grupo IVM. A resistência à IVM foi detectada, embora também se suspeitasse de resistência ao LV. O grupo IVM teve um tratamento eficaz significativamente maior, com 93,5%, resultando em um aumento (P < 0,05) do ganho de peso vivo de 16,1kg, seguido pelo grupo LV (92,4%), com 17,1kg, em comparação com o grupo controle sem tratamento. Foi observada uma correlação negativa (P < 0,05) entre o EPG e o ganho de peso para os grupos LV (r = -0,46) e IVM (r = -0,32). LV e IVM mostraram falta de eficácia contra nematódeos gastrointestinais, assim como uma capacidade adequada de redução de EPG, mas com resistência IVM e efeitos prejudiciais no desempenho de crescimento em gado de corte em pastagem.

Vasculitis inducida por cocaína-levamisol con manifestaciones cutáneas, articulares y abdominales: reporte de caso / Cocaine-levamisole induced vasculitis with cutaneous, joint and abdominal manifestations: case report

Resumen El levamisol es un antiparasitario de uso veterinario que actualmente es empleado para aumentar el volumen y la potencia de la cocaína. La mezcla de estas dos sustancias puede causar un cuadro caracterizado por lesiones propias de la cocaína, como la afección del cartílago septal con perforación del tabique nasal, y vasculitis cutánea de pequeños vasos con afectación de los pabellones auriculares y del cartílago nasal, afección conocida como vasculitis inducida por cocaína-levamisol (VICOL) que puede avanzar a necrosis e incluso ulceraciones cutáneas, asociadas a agranulocitosis, artralgias y glomerulonefritis . En el presente artículo se describe el caso de un paciente con historia de consumo de sustancias en quien se encontraron lesiones purpúricas palpables en miembros superiores, tronco, pabellones auriculares y miembros inferiores. Se consideró una clínica sugestiva de VICOL dado el antecedente de consumo de sustancias. En el proceso diagnóstico se descartaron entidades como la vasculitis por anticuerpos contra el citoplasma de los neutrófilos (ANCAs) y crioglobulinemia, entre otras posibles afecciones. Se llevó a cabo un tratamiento con esteroides y con ello presentó una respuesta adecuada, pero luego recurrieron los síntomas, particularmente abdominales, los cuales se consideraron asociados con vasculitis. Se le brindó manejo adicional con ciclofosfamida y nuevos pulsos de esteroides, con que se logró el control total de los síntomas. A través este caso se resaltan entonces los ejercicios diagnósticos y clínicos en la vasculitis cocaína- levamisol, y se sugiere la consideración de los síntomas abdominales como posible componente del cuadro vasculítico.

Abstract Levamisole is an antiparasitic agent for veterinary use. Currently it is used to increase the volume and potency of cocaine. Levamisole and cocaine combined result in the septum nasal perforation and small-vessel vasculitis in the ears and nasal cartilage. These findings are known as cocaine levamisole-induced vasculitis and can progress to necrosis and even skin ulceration, which is associated with agranulocytosis, arthralgia, and glomerulonephritis. This article describes the case of a patient with a history of substance abuse in whom palpable purpuric lesions were found in the upper and lower limbs, trunk, and ears. A clinical condition suggestive of vasculitis induced by cocaine-levamisole was considered, given the history of substance consumption. In the diagnostic process, entities such as Anti-neutrophil Cytoplasmic Antibodiy (ANCA) vasculitis and cryoglobulinemia, among other possible conditions, were ruled out. Steroid treatment was carried out, to which the patient had an adequate response, but then symptoms recurred, particularly abdominal, which were associated with vasculitis. Additional management with cyclophosphamide and new steroid pulses were provided, and with those symptom control was achieved. In this case report highlights the diagnostic and clinical exercises in cocaine levamisole vasculitis and is suggested the consideration of abdominal symptoms as a possible component of the vasculitis flare.

Co-administration of morphine and levamisole increases death risk, produces neutropenia and modifies antinociception in mice.

Levamisole is a veterinary anthelmintic drug and a common adulterant of misused drugs. This study analyses the lethal, antinociceptive and haematological effects produced by acute or repeated levamisole administration by itself or combined with morphine. Independent groups of male Swiss Webster mice were i.p. injected with 100 mg/kg morphine, 31.6 mg/kg levamisole (lethal doses at 10%, LD10 ) or the same doses combined. Naloxone pretreatment (10 mg/kg, i.p.) prevented morphine-induced death, as did 2.5 mg/kg, i.p. mecamylamine with levamisole. Co-administration of levamisole and morphine (Lvm + Mor) increased lethality from 10% to 80%. This augmented effect was prevented by 30 mg/kg, i.p. naloxone and reduced with 10 mg/kg naloxone plus 2.5 mg/kg, i.p. mecamylamine. In independent groups of mice, 17.7 mg/kg, i.p. levamisole antagonized the acute morphine's antinociceptive effect evaluated in the tail-flick test. Repeated 17.7 mg/kg levamisole administration (2×/day/3 weeks) did not affect tolerance development to morphine (10 mg/kg, 3×/day/1 week). Blood samples obtained from mice repeatedly treated with levamisole showed leukopenia and neutropenia. Morphine also produced neutropenia, increased erythrocyte count and other related parameters (e.g. haemoglobin). Lvm + Mor had similar effects on leukocyte and neutrophil counts to those seen with levamisole only, but no erythrocyte-related alterations were evident. Blood chemistry analysis did not indicate liver damage but suggested some degree of electrolyte balance impairment. In conclusion, Lvm + Mor increased death risk, altered morphine-induced antinociceptive effects and produced haematologic abnormalities. The importance of studying combinations of drugs of abuse lies in the fact that drug users frequently combine drugs, which are commonly adulterated.

Glomerulonefritis crescéntica secundaria a consumo de cocaína adulterada con levamisol. Reporte de un caso y revisión de la literatura / Crescentic glomerulonephritis associated with levamisole-adulterated cocaine. A case report and a review of the literature

Resumen Introducción: El levamisol es un fármaco antihelmíntico, también conocido por su uso como inmunomodulador el cual, como consecuencia de sus efectos tóxicos, fue retirado a finales del siglo XX. En el 2005, producto de un incremento en el diagnóstico de vasculitis pauci-inmune entre la población usuaria de sustancias psicoactivas, se documentó la adulteración con fines comerciales de la cocaína combinándola con levamisol, a partir de un aumento de manifestaciones reumáticas asociadas al consumo de dicha droga. Reporte de caso: Se presenta el caso de un adulto joven con antecedentes de síndrome de Alport y consumo reciente de sustancias psicoactivas. Se realiza biopsia renal demostrándose la presencia de glomerulonefritis crescéntica pauci-inmune. Por lo anterior, se relacionó este tipo de vasculitis de pequeño vaso con afectación renal y depósito de complejos inmunes al consumo de cocaína adulterada con levamisol. Discusión: El levamisol, medicamento aprobado por la FDA en 1991, actúa como inmunomodulador, antiparasitario y coadyuvante en quimioterapia. El levamisol produce un síndrome reumático caracterizado por la presencia de glomerulonefritis, hemorragia alveolar, púrpura retiforme, neutropenia y agranulocitosis en relación con la presencia de anticuerpos anticitoplasma de neutrófilo. Conclusión: El levamisol es conocido por sus propiedades antihelmínticas e inmunomoduladoras, adicionalmente puede producir efectos tóxicos ostensibles. Dado el alto consumo de cocaína entre la población indigente, la presencia de este adulterante constituye un problema de salud pública creciente.

Abstract Introduction: Levamisole is an anthelmintic drug, also known for its use as an immunomodulator which, because of its toxic effects, was withdrawn at the end of the 20th century. In 2005, because of an increase in the diagnosis of pauci-immune vasculitis among the population that uses psychoactive substances, the adulteration of cocaine for commercial purposes by combining it with levamisole was documented. Case Report: The case of a young adult with a history of Alport Syndrome and recent consumption of psychoactive substances is presented. A renal biopsy is performed, demonstrating the presence of pauci-immune crescentic glomerulonephritis. Therefore, this type of small vessel vasculitis with kidney involvement and immune complex deposition was associated with the use of cocaine adulterated with levamisole. Discussion: Levamisole, a drug approved by the FDA in 1991, acts as an immunomodulator, antiparasitic and adjuvant in chemotherapy. Levamisole produces a rheumatic syndrome characterized by the presence of glomerulonephritis, alveolar hemorrhage, retinal purpura, neutropenia, and agranulocytosis in association with the presence of anti-neutrophil cytoplasmic antibodies. Conclusion: levamisole is known for its anthelmintic and immunomodulatory properties, additionally it can produce ostensible toxic effects. Given the high consumption of cocaine among the indigent population, the presence of this adulterant constitutes a growing public health problem.

Pyoderma gangrenosum associated to the use of cocaine/levamisole. Series of three cases and literature review

Abstract Pyoderma gangrenosum associated to the use of cocaine/levamisole is a rare condition associated to their consumption. Cocaine use is frequent in Colombia, and the substance is contaminated with levamisole, an anthelmintic that increases the psychotropic effects and enhances its side effects. We present three clinical cases of patients with ulcerated lesions, in which the diagnosis was pyoderma gangrenosum secondary to the use of cocaine contaminated with levamisole. This called the attention of the health staff to investigate the abuse of substances in gangrenous pyoderma and also evidence that the interruption of consumption was the basis of management.

Pyoderma gangrenosum associated to the use of cocaine/levamisole. Series of three cases and literature review.

Pyoderma gangrenosum associated to the use of cocaine/levamisole is a rare condition associated to their consumption. Cocaine use is frequent in Colombia, and the substance is contaminated with levamisole, an anthelmintic that increases the psychotropic effects and enhances its side effects. We present three clinical cases of patients with ulcerated lesions, in which the diagnosis was pyoderma gangrenosum secondary to the use of cocaine contaminated with levamisole. This called the attention of the health staff to investigate the abuse of substances in gangrenous pyoderma and also evidence that the interruption of consumption was the basis of management.

Combined moxidectin-levamisole treatment against multidrug-resistant gastrointestinal nematodes: A four-year efficacy monitoring in lambs.

Nematicide combinations may be a valid strategy to achieve effective nematode control in the presence of drug resistance. The goal of the current trial was to evaluate the pharmaco-parasitological performance of the moxidectin (MOX) and levamisole (LEV) combination after four years of continuous use in lambs naturally parasitized with multi-resistant gastrointestinal nematodes. At the beginning of the trial, 40 lambs were divided into four groups (n = 10), which were untreated (control) or subcutaneously treated with MOX (0.2 mg/kg), LEV (8 mg/kg) or with the combination MOX + LEV (administered separately at 0.2 and 8 mg/kg, respectively). Blood samples were collected at different times post-treatment and LEV and MOX plasma concentrations were measured by HPLC. The clinical efficacy of the continuous use of MOX + LEV combination was assessed with the controlled efficacy test (CET), performed at the beginning and end of the study, and with the faecal egg count reduction (FECR) test, performed over the four-year study period. No significant adverse pharmacokinetic changes were observed either for MOX or LEV after their co-administration to infected lambs. The CET (first year) showed efficacies of 84.3 % (Haemonchus contortus), 100 % (Teladorsagia circumcincta and Trichostrongylus axei), and 97.4 % (T. colubriformis). After the repetitive use of the combined treatment for four years, those efficacies remained high (100 %) and only decreased to 58 % against T. colubriformis. The evaluation of the FECR over the study period showed fluctuations in the performance of the combined administration. The initial FECR (2014) was 99 % (MOX), 85 % (LEV) and 100 % (MOX + LEV). The co-administration of MOX + LEV during the four-year experimental period resulted in a significantly higher anthelmintic effect (87 %) than that of MOX (42 %) or LEV (69 %) given alone. The combined use of MOX + LEV to control resistant gastrointestinal nematodes appears to be a valid strategy under specific management conditions. A high initial therapeutic response to the combination would be a relevant feature for the success of this tool.

Combination of synthetic anthelmintics and monoterpenes: Assessment of efficacy, and ultrastructural and biophysical properties of Haemonchus contortus using atomic force microscopy.

The resistance of Haemonchus contortus to synthetic anthelmintics is of increasing concern; and different strategies are being evaluated to improve parasite control. The present study investigated the in vitro effects of combinations of synthetic compounds and monoterpenes. Additionally, the chemical association of the best combinations and their impact on the ultrastructural and biophysical properties of H. contortus eggs was evaluated. We assessed the efficacy of the monoterpenes, carvacrol, thymol, r-carvone, s-carvone, citral, and p-cymene and the anthelmintics, albendazole and levamisole using the egg hatch test (EHT) and the larval migration inhibition test (LMIT), respectively. The minimum effective concentrations of the monoterpenes, according to the EHT (efficacy ranging from 4.4%-11.8%) and LMIT (efficacy ranging from 5.6%-7.4%), were used in combination with different concentrations of synthetic compounds, and the IC50 and synergism rate (SR) were calculated. Fourier-transform infrared spectroscopy (FTIR) was used to analyze the chemical association between the best combinations as revealed by the in vitro tests (albendazole and levamisole with r-carvone or s-carvone). Atomic force microscopy (AFM) was used to assess the ultrastructural and biophysical properties of H. contortus eggs treated with the albendazole and r-carvone combination. Among the monoterpenes, the highest efficacies were exhibited by carvacrol (IC50 = 185.9 µg/mL) and thymol (IC50 = 187.0 µg/mL), according to the EHT, and s-carvone and carvacrol (IC50 = 1526.0 and 1785.3 µg/mL, respectively), according to the LMIT. According to the EHT, albendazole showed a slight statistically significant synergism in combination with r-carvone (SR = 3.8) and s-carvone (SR = 3.0). According to the LMIT, among the monoterpenes, r-carvone (SR = 1.7) and s-carvone (SR = 1.7) showed an increase in efficacy with levamisole; however, this was not statistically significant. The FTIR spectra of albendazole and levamisole, in association with r-carvone and s-carvone, indicated the presence of chemical interactions between the synthetic and natural molecules, contributing to the possible synergistic effects of these associations. Eggs treated with albendazole and r-carvone showed an increase in roughness and a decrease in height, suggesting that the treatment induced damage to the egg surface and an overflow of its internal contents. Overall, the combination of albendazole with r-carvone and s-carvone was efficacious against H. contortus, demonstrating a chemical association between the compounds; the significant changes in the egg ultrastructure justify this efficacy.

Clinical effectiveness of autohemotherapy as an adjuvant in the control of gastrointestinal nematodes in naturally infected sheep / Eficácia clínica da auto-hemoterapia como adjuvante no controle de nematódeos gastrointestinais em ovinos naturalmente infectados

This study aimed to evaluate the effects of autohemotherapy as an adjuvant in the control of gastrointestinal nematodes in sheep. Four experimental groups were formed: G1, 10 animals receiving autologous venous blood; G2, 10 animals receiving autologous venous blood and vermifuge containing levamisole; G3, 10 animals receiving only vermifuge containing levamisole; and G4, 10 animals as the control group receiving no treatment. We performed fecal egg count (eggs per gram, EPG) of strongyles, larval culture, hemogram, leukogram, and serum protein dosage prior to the start of treatment (D0), and on days 14 (D14) and 42 (D42). There was a significant decrease in the EPG of the groups receiving levamisole (G2 and G3) from D14 to the end of the experimental period. At the end of the evaluations, the mean EPG of G2 and G3 was significantly lower than that of G1 and G4. The most common nematode genus was Haemonchus (88%), and the least common was Trichostrongylus (1%). The Fecal Egg Count Reduction Test (FECRT) of G2 and G3 on D14 were 98.1% and 97.9%, respectively, however, in G1, the FECRT was zero on the two days when evaluation took place. G1 and G2 showed a significant increase in monocyte counts on D14 and D42. There was a significant increase in hematocrit and hemoglobin values in G2 and G3, however, a significant increase in the absolute value of red blood cells was observed only in G2. Two doses of autohemotherapy at 21-day intervals, administered alone or as an adjuvant to levamisole, is ineffective in controlling gastrointestinal nematodes in naturally infected sheep.

Este estudo objetivou avaliar os efeitos da auto-hemoterapia como adjuvante no controle de nematódeos gastrointestinais em ovinos. Quatro grupos experimentais foram formados: G1, 10 animais que receberam sangue venoso autólogo; G2, 10 animais que receberam sangue venoso autólogo e vermífugo contendo levamisol; G3, 10 animais que receberam somente vermífugo contendo levamisol; e G4, 10 animais do grupo controle, que não receberam tratamento. Realizamos contagem de ovos nas fezes (ovos por grama, OPG) de estrongilídeos, cultivo de larvas, hemograma, leucograma e dosagem de proteína sérica antes do início do tratamento (D0) e nos dias 14 (D14) e 42 (D42). Houve uma diminuição significativa no OPG dos grupos que receberam levamisole (G2 e G3) do D14 até o final do período experimental. Ao final das avaliações, o OPG médio de G2 e G3 foi significativamente menor do que G1 e G4. O gênero de nematódeo mais comumente encontrado foi Haemonchus (88%) e o menos foi Trichostrongylus (1%). O teste de Redução na Contagem de Ovos nas Fezes (RCOF) de G2 e G3 no D14 foi 98,1% e 97,9%, respectivamente, entretanto, no G1, o RCOF foi zero nos dois dias avaliados. G1 e G2 mostraram aumento significativo na contagem de monócitos em D14 e D42. Houve um aumento significativo nos valores do hematócrito e hemoglobina em G2 e G3, entretanto, um aumento significativo no valor absoluto de hemácias foi observado somente em G2. Duas doses de auto-hemoterapia em intervalos de 21 dias, administradas isoladamente ou como adjuvante do levamisole, não é eficaz no controle de nematóides gastrintestinais em ovinos naturalmente infectados.

Eficácia clínica da auto-hemoterapia como adjuvante no controle de nematódeos gastrointestinais em ovinos naturalmente infectados / Clinical effectiveness of autohemotherapy as an adjuvant in the control ofgastrointestinal nematodes in naturally infected sheep

Este estudo objetivou avaliar os efeitos da auto-hemoterapia como adjuvante no controle de nematódeos gastrointestinais em ovinos. Quatro grupos experimentais foram formados: G1, 10 animais que receberam sangue venoso autólogo; G2, 10 animais que receberam sangue venoso autólogo e vermífugo contendo levamisol; G3, 10 animais que receberam somente vermífugo contendo levamisol; e G4, 10 animais do grupo controle, que não receberam tratamento. Realizamos contagem de ovos nas fezes (ovos por grama, OPG) de estrongilídeos, cultivo de larvas, hemograma, leucograma e dosagem de proteína sérica antes do início do tratamento (D0) e nos dias 14 (D14) e 42 (D42). Houve uma diminuição significativa no OPG dos grupos que receberam levamisole (G2 e G3) do D14 até o final do período experimental. Ao final das avaliações, o OPG médio de G2 e G3 foi significativamente menor do que G1 e G4. O gênero de nematódeo mais comumente encontrado foi Haemonchus (88%) e o menos foi Trichostrongylus (1%). O teste de Redução na Contagem de Ovos nas Fezes (RCOF) de G2 e G3 no D14 foi 98,1% e 97,9%, respectivamente, entretanto, no G1, o RCOF foi zero nos dois dias avaliados. G1 e G2 mostraram aumento significativo na contagem de monócitos em D14 e D42. Houve um aumento significativo nos valores do hematócrito e hemoglobina em G2 e G3, entretanto, um aumento significativo no valor absoluto de hemácias foi observado somente em G2. Duas doses de auto-hemoterapia em intervalos de 21 dias, administradas isoladamente ou como adjuvante do levamisole, não é eficaz no controle de nematóides gastrintestinais em ovinos naturalmente infectados.

This study aimed to evaluate the effects of autohemotherapy as an adjuvant in the control of gastrointestinal nematodes in sheep. Four experimental groups were formed: G1, 10 animals receiving autologous venous blood; G2, 10 animals receiving autologous venous blood and vermifuge containing levamisole; G3, 10 animals receiving only vermifuge containing levamisole; and G4, 10 animals as the control group receiving no treatment. We performed fecal egg count (eggs per gram, EPG) of strongyles, larval culture, hemogram, leukogram, and serum protein dosage prior to the start of treatment (D0), and on days 14 (D14) and 42 (D42). There was a significant decrease in the EPG of the groups receiving levamisole (G2 and G3) from D14 to the end of the experimental period. At the end of the evaluations, the mean EPG of G2 and G3 was significantly lower than that of G1 and G4. The most common nematode genus was Haemonchus (88%), and the least common was Trichostrongylus (1%). The Fecal Egg Count Reduction Test (FECRT) of G2 and G3 on D14 were 98.1% and 97.9%, respectively, however, in G1, the FECRT was zero on the two days when evaluation took place. G1 and G2 showed a significant increase in monocyte counts on D14 and D42. There was a significant increase in hematocrit and hemoglobin values in G2 and G3, however, a significant increase in the absolute value of red blood cells was observed only in G2. Two doses of autohemotherapy at 21-day intervals, administered alone or as an adjuvant to levamisole, is ineffective in controlling gastrointestinal nematodes in naturally infected sheep.

Agranulocitosis febril y artropatía reactiva asociada a consumo de cocaína en un paciente HLA B27 positivo / Febrile agranulocytosis and reactive arthropathy associated with cocaine consumption in an HLA B27 positive patient

Resumen La agranulocitosis asociada al consumo de cocaína es un fenómeno vinculado a la presencia de levamisol, un agente antihelmíntico e inmunomodulador, usado como adulterante de la cocaína. Esta reacción puede presentarse con mayor frecuencia en personas con HLA B27. Además de la agranulocitosis, las personas que consumen cocaína adulterada con levamisol pueden desarrollar fiebre, lesiones en piel, artralgias y, menos frecuentemente, artritis y entesitis inflamatoria. Presentamos el caso de un paciente consumidor de cocaína, con genotipo HLA B27, que desarrolló agranulocitosis febril y artropatía reactiva. En sangre se detectó la presencia de ANCA p, ANCA atípico y MPO, y fueron excluidas otras causas de agranulocitosis. Fue tratado con corticoides y posteriormente metotrexato, terapia de deshabituación, con buena evolución.

Abstract Agranulocytosis associated with cocaine use is a phenomenon linked to the presence of levamisole, an anthelminthic and immunomodulating agent, used as an adulterant to cocaine. This reaction has been associated with the presence of HLA B27. In addition to agranulocytosis, people who use levamisole-adulterated cocaine may develop fever, skin lesions, arthralgias, and less frequently, inflammatory enthesitis and arthritis. We present the case of a cocaine-consuming patient with HLA B27 genotype, who developed febrile agranulocytosis and inflammatory arthropathy. The presence of p ANCA, atypical ANCA and MPO was detected in blood, and other causes of agranulocytosis were excluded. He was treated with corticosteroids and later methotrexate, therapy for addiction, with good evolution.

[Febrile agranulocytosis and reactive arthropathy associated with cocaine consumption in an HLA B27 positive patient]. / Agranulocitosis febril y artropatía reactiva asociada a consumo de cocaína en un paciente HLA B27 positivo.

Agranulocytosis associated with cocaine use is a phenomenon linked to the presence of levamisole, an anthelminthic and immunomodulating agent, used as an adulterant to cocaine. This reaction has been associated with the presence of HLA B27. In addition to agranulocytosis, people who use levamisole-adulterated cocaine may develop fever, skin lesions, arthralgias, and less frequently, inflammatory enthesitis and arthritis. We present the case of a cocaine-consuming patient with HLA B27 genotype, who developed febrile agranulocytosis and inflammatory arthropathy. The presence of p ANCA, atypical ANCA and MPO was detected in blood, and other causes of agranulocytosis were excluded. He was treated with corticosteroids and later methotrexate, therapy for addiction, with good evolution.

La agranulocitosis asociada al consumo de cocaína es un fenómeno vinculado a la presencia de levamisol, un agente antihelmíntico e inmunomodulador, usado como adulterante de la cocaína. Esta reacción puede presentarse con mayor frecuencia en personas con HLA B27. Además de la agranulocitosis, las personas que consumen cocaína adulterada con levamisol pueden desarrollar fiebre, lesiones en piel, artralgias y, menos frecuentemente, artritis y entesitis inflamatoria. Presentamos el caso de un paciente consumidor de cocaína, con genotipo HLA B27, que desarrolló agranulocitosis febril y artropatía reactiva. En sangre se detectó la presencia de ANCA p, ANCA atípico y MPO, y fueron excluidas otras causas de agranulocitosis. Fue tratado con corticoides y posteriormente metotrexato, terapia de deshabituación, con buena evolución.

Impact of levamisole in co-administration with benznidazole on experimental Chagas disease.

Levamisole (Lms) is an anthelminthic drug with immunomodulatory activity. Chagas disease (CD) is caused by Trypanosoma cruzi and there is very low access to the drugs available, benznidazole (Bz) and nifurtimox, both far from ideal. In a drug-repurposing strategy to test potential activity as antiparasitic and immunomodulatory agent for CD, Lms was assayed on acute T. cruzi murine infection, alone and in co-administration with Bz. During protocol standardization, 100 and 10 mpk of Bz given for five consecutive days resulted in parasitaemia suppression and 100% animal survival only with the highest dose. Flow cytometry showed that both optimal (100 mpk) and suboptimal (10 mpk) doses of Bz equally decreased the plasma levels of cytokines commonly elevated in this acute infection model. Lms alone (10-0.5 mpk) did not decrease parasitaemia nor mortality rates. Co-administration was investigated using the suboptimal dose of Bz and different doses of Lms. While Bz 10 mpk did not alter parasitaemia, the combo partially reduced it but only slightly promoted animal survival. This effect could be related to Th1-response modulation since interleukin-6 and interferon-γ were higher after treatment with the combo.

Pharmaco-parasitological evaluation of the ricobendazole plus levamisole nematodicidal combination in cattle.

The goals of the current study were to evaluate the potential pharmacokinetic (PK) interactions and the clinical efficacy occurring after the subcutaneous (s.c.) administration of ricobendazole (RBZ) and levamisole (LEV) given both separately and co-administered to calves naturally infected with susceptible gastrointestinal nematodes. The clinical efficacy was shown in two seasons, winter and spring, with predominance of different nematode populations. Groups of 15 calves were treated with RBZ alone, LEV alone and RBZ + LEV combination, and an untreated group was kept as a Control. RBZ and LEV plasma concentrations were quantified by HPLC. The clinical efficacy was determined by the faecal egg count reduction test. RBZ and LEV have similar plasma persistence, being detected in plasma over 24 hr post-treatment. No PK interactions were observed after the combined treatment, with similar PK parameters (p > .05) obtained for the single-drug and the combination-based strategy. In winter, the observed clinical efficacies were 96%, 99% and 100% for groups treated with RBZ, LEV and RBZ + LEV, respectively; however, in spring, the efficacies were 95%, 93% and 96% for the same groups. Remarkably, the combination was the only treatment that achieved 100% clinical efficacy against both Haemonchus spp and Ostertagia spp in winter; but the increased presence of Ostertagia spp. in spring (28% in untreated group) determined a tendency to reduced efficacies compared to winter time (only 10% of Ostertagia spp. in untreated group), even for the combined treatment. Overall, in a scenario where the nematode population is susceptible, the RBZ + LEV treatment may be a valid combination in cattle to delay the development of resistance, especially in winter when this combination achieved 100% of efficacy. Thus, selection of anthelmintic resistance will never occur. In fact, this is one of the greatest challenges for the whole cattle production system: to be one step ahead of anthelmintic resistance.

In vitro susceptibility of nematophagous fungi to antiparasitic drugs: interactions and implications for biological control

Abstract The fast anthelmintic resistance development has shown a limited efficiency in the control of animals endoparasitosis and has promoted research using alternative control methods. The use of chemicals in animal anthelmintic treatment, in association with nematophagous fungi used for biological control, is a strategy that has proven to be effective in reducing the nematode population density in farm animals. This study aims to verify the in vitro susceptibility of the nematophagous fungi Arthrobotrys oligospora, Duddingtonia flagrans and Paecilomyces lilacinus against the antiparasitic drugs albendazole, thiabendazole, ivermectin, levamisole and closantel by using the Minimum Inhibitory Concentration (MIC). MICs ranged between 4.0 and 0.031 µg/mL for albendazole, thiabendazole and ivermectin, between 0.937 and 0.117 µg/mL for levamisole, and between 0.625 and 0.034 µg/mL for closantel. The results showed that all antiparasitic drugs had an in vitro inhibitory effect on nematophagous fungi, which could compromise their action as agents of biological control. D. flagrans was the most susceptible species to all drugs.

Resumo O desenvolvimento rápido da resistência anti-helmíntica demonstrou a eficiência limitada no controle de endoparasitoses em animais, e promoveu a investigação em métodos de controles alternativos. O uso de produtos químicos no tratamento anti-helmíntico animal, em associação com fungos nematófagos utilizados para o controlo biológico, é uma estratégia que tem provado ser eficaz na redução da densidade da população de nematódeos em animais agrícolas. Este estudo teve como objetivo verificar a suscetibilidade in vitro dos fungos nematófagos Arthrobotrys oligospora, Duddingtonia flagrans e Paecilomyces lilacinus frente aos antiparasitários albendazol, tiabendazol, ivermectina, levamisol e closantel, usando a concentração inibitória mínima (MIC). Os MICs variaram entre 4,0 e 0,031 g/mL para albendazol, tiabendazol e ivermectina, entre 0,937 e 0,117 g/mL para o levamisol, e entre 0,625 e 0,034 g/mL para closantel. Os resultados mostraram que todos os antiparasitários tiveram um efeito inibidor in vitro sobre os fungos nematófagos, o que poderia comprometer suas atividades como agentes de controle biológico. D. flagrans foi a espécie mais sensível a todas as drogas.

In vitro susceptibility of nematophagous fungi to antiparasitic drugs: interactions and implications for biological control / Suscetibilidade in vitro de fungos nematófagos à antiparasitários: interações e implicações para o controle biológico

Abstract The fast anthelmintic resistance development has shown a limited efficiency in the control of animal's endoparasitosis and has promoted research using alternative control methods. The use of chemicals in animal anthelmintic treatment, in association with nematophagous fungi used for biological control, is a strategy that has proven to be effective in reducing the nematode population density in farm animals. This study aims to verify the in vitro susceptibility of the nematophagous fungi Arthrobotrys oligospora, Duddingtonia flagrans and Paecilomyces lilacinus against the antiparasitic drugs albendazole, thiabendazole, ivermectin, levamisole and closantel by using the Minimum Inhibitory Concentration (MIC). MICs ranged between 4.0 and 0.031 µg/mL for albendazole, thiabendazole and ivermectin, between 0.937 and 0.117 µg/mL for levamisole, and between 0.625 and 0.034 µg/mL for closantel. The results showed that all antiparasitic drugs had an in vitro inhibitory effect on nematophagous fungi, which could compromise their action as agents of biological control. D. flagrans was the most susceptible species to all drugs.

Resumo O desenvolvimento rápido da resistência anti-helmíntica demonstrou a eficiência limitada no controle de endoparasitoses em animais, e promoveu a investigação em métodos de controles alternativos. O uso de produtos químicos no tratamento anti-helmíntico animal, em associação com fungos nematófagos utilizados para o controlo biológico, é uma estratégia que tem provado ser eficaz na redução da densidade da população de nematódeos em animais agrícolas. Este estudo teve como objetivo verificar a suscetibilidade in vitro dos fungos nematófagos Arthrobotrys oligospora, Duddingtonia flagrans e Paecilomyces lilacinus frente aos antiparasitários albendazol, tiabendazol, ivermectina, levamisol e closantel, usando a concentração inibitória mínima (MIC). Os MICs variaram entre 4,0 e 0,031 μg/mL para albendazol, tiabendazol e ivermectina, entre 0,937 e 0,117 μg/mL para o levamisol, e entre 0,625 e 0,034 μg/mL para closantel. Os resultados mostraram que todos os antiparasitários tiveram um efeito inibidor in vitro sobre os fungos nematófagos, o que poderia comprometer suas atividades como agentes de controle biológico. D. flagrans foi a espécie mais sensível a todas as drogas.

Cocaine-Levamisole-Induced Vasculitis/Vasculopathy Syndrome.

PURPOSE OF REVIEW: To understand the clinical spectrum of cocaine-levamisole-induced vasculitis. Worldwide recreational drug consumption is high among the adult population from various social strata. The use of cocaine with levamisole, a frequently added antiparasitic diluent, favors the manifestations of vasculitic lesions, especially in the skin. RECENT FINDINGS: New insights into immunological mechanisms involved in the pathogenesis of the disease. There are still many unknown aspects in the pathogenesis of this disease, such as the immune system interaction with p-ANCAs and the release of inflammatory NETs (neutrophil extracellular traps), which are the origin of auto-antigens and tissue damage, manifesting as vasculitic purpura on the skin. The clinical presentation constitutes a challenge for the clinician to be able to distinguish it from small-vessel vasculitides. This paper intends to improve the understanding of this condition, exhibiting the broad clinical spectrum of local and systemic manifestations of cocaine-levamisole-induced vasculitis, to facilitate a timely diagnosis, in order to take corrective measures and avoid sequelae, along with tissue damage and the consequent deformities and permanent scars.

Anthelmintic effect of Psidium guajava and Tagetes erecta on wild-type and Levamisole-resistant Caenorhabditis elegans strains.

ETHNOPHARMACOLOGICAL RELEVANCE: Psidium guajava and Tagetes erecta have been used traditionally to treat gastrointestinal parasites, but their active metabolites and mechanisms of action remain largely unknown. AIM OF THE STUDY: To evaluate the anthelmintic potential of Psidium guajava and Tagetes erecta extracts on Levamisole-sensitive and Levamisole-resistant strains of the model nematode Caenorhabditis elegans. MATERIALS AND METHODS: Aqueous extracts of Psidium guajava (PGE) and Tagetes erecta (TEE) were assayed on locomotion and egg-laying behaviors of the wild-type (N2) and Levamisole-resistant (CB193) strains of Caenorhabditis elegans. RESULTS: Both extracts paralyzed wild-type and Levamisole-resistant nematodes in a dose-dependent manner. In wild-type worms, TEE 25mg/mL induced a 75% paralysis after 8h of treatment and PGE 25mg/mL induced a 100% paralysis after 4h of treatment. PGE exerted a similar paralyzing effect on N2 wild-type and CB193 Levamisole-resistant worms, while TEE only partially paralyzed CB193 worms. TEE 25mg/mL decreased N2 egg-laying by 65% with respect to the untreated control, while PGE did it by 40%. CONCLUSIONS: Psidium guajava leaves and Tagetes erecta flower-heads possess hydrosoluble compounds that block the motility of Caenorhabditis elegans by a mechanism different to that of the anthelmintic drug Levamisole. Effects are also observable on oviposition, which was diminished in the wild-type worms. The strong anthelmintic effects in crude extracts of these plants warrants future work to identify their active compounds and to elucidate their molecular mechanisms of action.