Alfredo Skrebsky Cezar

Multiple resistance of gastrointestinal nematodes to nine different drugs in a sheep flock in southern Brazil

Based on clinical observation of a flock of approximately 5000 sheep (breed Merino) from southern Brazil, the failure of anthelmintic treatments was suspected. The sensitivity of the gastrointestinal nematodes that infected these sheep to nine drugs (Levamisole, Moxidectin, Albendazole, Ivermectin, Nitroxynil, Disophenol, Trichlorphon, Closantel, and a combination of Ivermectin, Levamisole and Albendazole) was evaluated. None of the drugs reduced the count of nematode eggs per gram of feces (EPG) in the sheep to an adequate extent (defined as a 95% reduction in EPG). Therefore, we found multiple parasite resistance to all classes of anthelmintic drugs available for small ruminants in the Brazilian market. The genera Haemonchus spp., Trichostrongylus spp. and Ostertagia spp. had various degrees of resistance (or natural tolerance) to each of the drug treatments.

Controle alternativo de nematódeos gastrintestinais dos ruminantes: atualidade e perspectivas

Infections by gastrointestinal nematodes cause significant losses in the production of ruminants. Intensification of production process tends to enhance the effects of parasitosis. Easy access for producers to anthelmintics, in addition to poor professional assistance, has led to indiscriminate use of these chemicals and, consequently, to parasite resistance, reducing the effectiveness of treatment. Compounding the issue there is a trend towards valorization of organic products, originating from production systems where the use of chemotherapics is restricted or prohibited. This context emphasizes the importance of alternative approaches based on: management of livestock and pastures, biological control of parasites, stimulation of immunological response of host and phytotherapy. There are some promising methods by future utilization and others viable by immediate application, which, combined with conventional anthelmintics, can guarantee success and sustainability in controlling the nematodiasis.

Ação anti-helmíntica de diferentes formulações de lactonas macrocíclicas em cepas resistentes de nematódeos de bovinos

The macrocyclic lactones (MLs) (avermectins and milbemycins) are endectocides broadly used in livestock and in some parasitic diseases of humans. In cattle, parasite resistance to MLs is emerging, and the appearance of formulations that differ in their pharmacological properties become complex the choice of the most appropriate drug to each case. In order to evaluate possible alternatives to restore the effectiveness of MLs on resistant strains of gastrointestinal nematodes, were tested, in this study, ten different treatments based on the MLs on a population of gastrointestinal nematodes of cattle which, known, was under pressure of selection by 1% avermectins. Additionally, was tested a benzimidazole. The efficacy of the drugs was calculated with basis on the reduction of eggs per gram of feces (EPG) of cattle. The resistance of each genus was evaluated by identification of the larvae, obtained from culture in the feces, pre- and post-treatments. The desired efficacy was not obtained using long action avermectins - with high concentration and in association - even with the application of high doses. The genera Cooperia spp., Haemonchus spp. and Trichostrongylus spp. were resistant to avermectins, and Ostertagia spp. to ivermectin. It was observed that, once established parasite resistance to the 1% MLs, the application of drugs, of this same chemical group, even in formulations of high concentration, association or in high doses, may not result in the expected efficacy.

Neosporose equina: ocorrência de anticorpos anti-Neospora spp. e associação entre status sorológico de éguas e de suas crias

Neospora caninum and N. hughesi are protozoa which can infect horses and can cause reproductive and neurological diseases, respectively. The pathogenesis of neosporosis in horses is poorly understood, as well as the sources of horizontal infection of N. hughesi. Furthermore, there are doubts about the role of the vertical transmission of Neospora spp. in maintenance of these parasites in equine populations. In this study, we evaluated: (1) the occurrence of infections by Neospora spp. in a population of mares (in reproductive age) on a farm of Crioula breed horses; and (2) the possible association between the serological status of mares and of their offspring, aiming to investigate, indirectly, the relevance of transplacental transmission for the occurrence of Neospora spp. in these horses. We found a highly significant association between the serological status of mares and their offspring. Although had been exposed to the same environmental risk factors, the descendants of seropositive mares had a higher percentage of seropositivity against Neospora spp. compared to the descendants of seronegative mares. The association between kinship and serological status indicates an influence of vertical (transplacental) infection raising the occurrence of Neospora spp. in the studied equine population.

Combinação de drogas antiparasitárias como uma alternativa para o controle de nematódeos gastrintestinais multirresistentes em ovinos

This study was conducted to find effective anthelmintic treatments on a multi-resistant population of gastrointestinal nematodes of sheep, which was found resisting to all the classes of nematicidal drugs available in the Brazilian market. We took as a starting point the data obtained in a previous study performed in the targeted flock, in which were tested conventional treatments. Thus, using the calculated efficacy of the drugs (tested previously) for the genera of nematodes found in the flock, we selected drugs which, combined or used in higher doses, could improve the efficacy of the treatments. We evaluated the anthelmintic action of each treatment using the percentage of reduction of eggs in the feces, along with larvae cultures. A combination of moxidectin 1% + disophenol 20% showed 99% of efficacy, and combinations of moxidectin 1% + trichlorphon 10% and moxidectin 1% + levamisole phosphate 22.3% showed efficacy over 90%. Combinations of drugs, with different mechanisms of action, can provide effective anthelmintic treatments on multidrug-resistant populations of gastrointestinal nematodes of sheep.

Comparative performance of macrocyclic lactones against large strongyles in horses

Several formulations of macrocyclic lactones (abamectin, ivermectin, moxidectin), including ivermectin combined with pyrantel (tetrahydropyrimidine) and ivermectin combined with praziquantel (pyrazinoisoquinolin derivative), were tested regarding their efficacy to control gastrointestinal nematodes of horses on a stud farm in southern Brazil. In addition, we tested a pharmaceutically produced generic paste containing ivermectin 4%. Similar formulations of avermectins had different efficacies measured by reduction of EPG. Levels of efficacy of the tested drugs varied against Strongylus edentatus, S. equinus and S. vulgaris. The generic paste (ivermectin 4%) was less effective than the conventional drugs.

Detecção de anticorpos anti-Neospora caninum em amostras individuais e coletivas de leite de bovinos pela reação de imunofluorescência indireta

Neospora caninum is a major causative agent of reproductive losses in cattle and the diagnosis of neosporosis is essential for control and eradication programs. Therefore, this study aimed to: (1) standardize an indirect immunofluorescence assay (IFAT) for detection of antibodies against N. caninum in bovine milk, starting from a standard IFAT used for blood serum samples; (2) check the agreement between antibodies detection by IFAT in blood serum and in milk of cows; (3) evaluate the suitability of IFAT for detection of anti-N. caninum antibodies in bulk milk samples. We tested samples of blood serum and milk collected from 112 lactating cows for detection of antibodies against N. caninum, furthermore, six bulk milk samples, each one corresponding to each dairy farm studied. Agreement between the detection of antibodies in blood serum (with antibody titer >50) and in milk, with 90% of sensitivity and 100% of specificity for the IFAT in milk samples were found in 78% of the animals. However, for cows with antibody titers >100 in blood serum, the agreement, the sensitivity and the specificity of the IFAT in milk were of 100%. It is shown that, considering the dairy farm conditions. The most appropriate diagnostic approach to be adopted regarding the collection of blood serum or milk can elect for search anti-N. caninum antibodies by IFAT. Moreover, detection of antibodies in bulk milk samples can serve for diagnosis and screening of herds with infected animals.

Occurrence of antibodies anti-Neospora spp. in cart horses and Crioula breed horses from Rio Grande do Sul state

O objetivo deste estudo foi avaliar a frequencia de deteccao de anticorpos contra Neospora spp. em cavalos de carroca e em cavalos da raca Crioula. Para tal, 214 amostras de soro foram coletadas e analisadas pela tecnica de imunofluorescencia indireta, das quais 91 eram de cavalos de carroca e 123 de cavalos da raca Crioula, todas provenientes da regiao central do Rio Grande do Sul. As frequencias de anticorpos detectadas foram: 15,9% (34/214) na populacao total estudada, 15,4% (14/91) nos cavalos de carroca e 16,3% (20/123) nos cavalos da raca Crioula. Estes resultados sugerem que a infeccao por Neospora spp. esta presente igualmente nas duas populac oes. Assim, devido a importância e ao padrao da patogenese da neosporose em equinos, Neospora spp. deve ser incluido no diagnostico de problemas reprodutivos em eguas e em casos de problemas neurologicos em equinos.

Anticorpos anti-Neospora caninum em bovinos de leite do sudoeste do estado do Paraná

Since November 2007 until May 2009, 1,778 serum samples of cattle from dairy herds of the Southwest of Parana State, Brazil, were used for search of anti-Neospora caninum antibodies. The frequency of seropositive animals, assessed by IFAT, was 24.2% (431/1,178), showing a relatively high occurrence in the studied population. These results show that Neospora caninum is widely distributed in the dairy cattle in the Southwest of Parana State. The presence of Neospora caninum in a herd is worrisome, since this protozoan is closely related with reproductive disorders and low milk production in cattle.

Effects of Adiponectin Including Reduction of Androstenedione Secretion and Ovarian Oxidative Stress Parameters In Vivo.

Adiponectin is the most abundantly produced human adipokine with anti-inflammatory, anti-oxidative, and insulin-sensitizing properties. Evidence from in vitro studies has indicated that adiponectin has a potential role in reproduction because it reduces the production of androstenedione in bovine theca cells in vitro. However, this effect on androgen production has not yet been observed in vivo. The current study evaluated the effect of adiponectin on androstenedione secretion and oxidative stress parameters in a rodent model. Seven-week-old female Balb/c mice (n = 33), previously treated with equine gonadotropin chorionic, were assigned to one of four different treatments: Group 1, control (phosphate-buffered saline); Group 2, adiponectin 0.1 μg/mL; Group 3, adiponectin 1.0 μg/mL; Group 4, adiponectin 5.0 μg/mL. After 24 h, all animals were euthanized and androstenedione levels were measured in the serum while oxidative stress markers were quantified in whole ovary tissue. Female mice treated with adiponectin exhibited a significant reduction (about 60%) in serum androstenedione levels in comparison to controls. Androstenedione levels decreased from 0.78 ± 0.4 ng/mL (mean ± SD) in controls to 0.28 ± 0.06 ng/mL after adiponectin (5 μg/mL) treatment (P = 0.01). This change in androgen secretion after 24 hours of treatment was associated with a significant reduction in the expression of CYP11A1 and STAR (but not CYP17A1). In addition, ovarian AOPP product levels, a direct product of protein oxidation, decreased significantly in adiponectin-treated mice (5 μg/mL); AOPP (mean ± SD) decreased to 4.3 ± 2.1 μmol/L in comparison with that of the controls (11.5 ± 1.7 μmol/L; P = 0.0003). Our results demonstrated for the first time that acute treatment with adiponectin reduced the levels of a direct oxidative stress marker in the ovary as well as decreased androstenedione serum levels in vivo after 24 h.