Andreia Volpato

Trichostrongylus and Haemonchus anthelmintic resistance in naturally infected sheep from southern Brazil

The anthelmintic resistance in small ruminants is a common problem and concern worldwide. The aim of this study was to verify anthelmintic treatment efficacy in naturally infected sheep. This study was conducted on nine herds that used the same anthelmintic management for over a year. In each farm, the animals were divided into two groups: untreated control group (n = 5) and treated (n = 10) according to the number of eggs per gram of feces (EPG). The treatment effect was checked based on EPG results and larval culture performed before treatment and 10 days after treatment. Significant differences were not observed (P > 0.05) on EPG results between untreated and treated groups. The coproculture showed that the animals were infected primarily by Haemonchus spp., Trichostrongylus spp., Teladorsagia spp., Cooperia spp. and Oesophagostomum spp. In all farms, anthelmintic resistance by genera Haemonchus and Trichostrongylus was found, but this resistance varied greatly between farms. Haemonchus spp. showed resistance to closantel, levamisole, and albendazole. Trichostrongylus spp. was shown to be resistant to closantel, levamisole, and albendazole. The drugs tested showed to be efficient against the genera Teladorsagia, Cooperia, and Oesophagostomum. Based on these results, we conclude that the anthelmintic resistance to the tested drugs is a problem present in the farms evaluated.

In vitro effect of seven essential oils on the reproduction of the cattle tick Rhipicephalus microplus.

Graphical abstract

Occurrence of gastrointestinal parasites in goats from the Western Santa Catarina, Brazil

This study aims to investigate the occurrence of gastrointestinal parasites in goats from the Western Santa Catarina, Brazil. Twenty four farms were analyzed in 17 different municipalities. Animals (n=217) from different production purposes (milk and meat) and age were randomly chosen. Fecal samples were collected from the rectum stored in plastic bottles and transported to the laboratory in portable coolers at 10 °C. The technique of centrifugal flotation with saturated sugar solution was carried out in order to investigate the presence of eggs, cysts, and oocysts of gastrointestinal parasites. In 88.9% of the investigated animals, it was observed that the presence of nematode eggs which belongs to the Strongylida order, after cultivation and larvae identification were identified as Haemonchus spp., Trichostrongylus spp., Teladorsagia spp., Cooperia spp., and Oesophagostomum spp. Eggs of Thysanosoma, Trichuris, Moniezia, and Neoascaris genus were also observed. Additionally, the presence of oocysts of Eimeria spp. and Cryptosporidium spp. as well as cysts of Giardia spp., and Entamoeba spp. were verified. In all the farms evaluated, the animals showed a single or mixed infection, with the highest occurrence of helminths belonging to the Haemonchus and Trichostrongylus genus, as well as the protozoan Eimeria.

Melaleuca alternifolia Essential Oil against the Lesser Mealworm (Alphitobius diaperinus) and Its Possible Effect on the Soil Fauna

The aim of this study was to evaluate the in vitro bioactivity of tea tree (Melaleuca alternifolia) essential oil against larvae and adult forms of lesser mealworms (Alphitobius diaperinus) and its influence on the soil fauna. Tests were performed in triplicate using pure tea tree oil (TTO; 1, 5, 10, 25, 50, and 100%), TTO nanoparticles (1, 3, and 7.5%), or terpinen-4-ol, the main compound of the tea tree oil, at the same concentrations of TTO. Larvae and adult mortality occurred at concentrations up to 10 and 50% of TTO, respectively. No larvicidal or insecticidal effect of TTO nanoparticles was observed. Terpinen-4-ol showed insecticidal and larvicidal effect at concentrations higher than 25%. The evaluation of TTO effect on soil organisms was performed by standard ecotoxicological tests (ISO) with the springtail species Folsomia candida. Only TTO was used for ecotoxicological tests in doses of 1, 5, 10, 25, 50, and 100 mg kg-1 of soil. TTO had no negative effects on F. candida survival or reproduction. Therefore, it was concluded that M. alternifolia oil may be a new alternative for control of the lesser mealworm.

Experimental infection by Trypanosoma evansi in sheep: Occurrence of transplacental transmission and mice infection by parasite present in the colostrum and milk of infected ewes.

The aims of this study were to evaluate vertical transmission of Trypanosoma evansi in sheep experimentally infected, in addition to the mammary transmission by colostrum or milk of these infected sheep to mice. Three pregnant sheep were used: one uninfected, four months pregnant (Sheep A); and two (Sheep B and C) infected intravenously by T. evansi trypomastigotes (4.6×10(6) per animal) on the third (Sheep C) and fourth (Sheep B) month of pregnancy. Both infected sheep developed low and oscillating parasitemia measured by blood smears. Hemogram was performed at seven day intervals, showing anemia, leukocytosis, and lymphocytosis on sheep B and C. Three sheep had twins, where sheep A delivered healthy lambs and both infected sheep had delivered at least one stillborn. Additionally, lambs from sheep B and C died 24 and 72 h post-partum, respectively. Before colostrum intake, four lambs from infected sheep were positives for T. evansi according to blood smear evaluation, serology (CATT/T. evansi), and PCR. Sheep colostrum and milk samples collected from the first four days post-partum were positives for T. evansi on PCR, and these samples were able to infect seven mice (out of 10) orally (n=4/5) and intraperitoneally (n=3/5). Therefore, we conclude that the vertical transmission of T. evansi occurs in pregnant sheep, in addition to a strong possibility of the transmission by colostrum and milk.

Influence of rosemary, andiroba and copaiba essential oils on different stages of the biological cycle of the tick Rhipicephalus microplus in vitro

The aim of this study was to verify in vitro the influence of rosemary, andiroba, and copaiba essential oils on different stages of the cattle tick Rhipicephalus microplus. Female ticks were collected from naturally infected cows and treated in vitro with these essential oils at concentrations of 5% and 10%. All tests were performed in triplicate using positive (amitraz 10%) and negative (untreated) controls. It was possible to observe that rosemary, andiroba, and copaiba essential oils inhibited female reproduction at concentrations of 5% (45.0%, 77.5%, and 71.6%, respectively) and 10% (55.7%, 92.0,% and 86.7%, respectively) compared with amitraz (72.9%). Additionally, andiroba (10%) and copaiba (5% and 10%) oil solutions also had an acaricidal effect (100%), similarly to the positive control. The andiroba and copaiba oils showed an ovicidal effect, which effected hatchability. Rosemary oil showed neither acaricidal nor ovicidal effect. Therefore, based on these results, we were able to show that all oils and concentrations tested may affect tick reproduction by inhibiting tick oviposition and hatchability. In addition, andiroba and copaiba oils exhibited ovicidal and acaricidal effects.

A prophylactic protocol to stimulate the immune response also controls infectious disease and, consequently, minimizes diarrhea in newborn heifers

The immunostimulatory and immunomodulatory properties of selenium (Se), an essential trace element for animals, has increase its use because may prevent/or reduce the occurrence of infectious diseases. Thus, the aim of this study was to verify whether Se and vitamins (A and E) applied via subcutaneous associated with secnidazole via oral exert positive effects in the antioxidant and immune systems, as well as whether prevent infections caused by protozoan and bacteria, and consequently, reduce the number of cases of diarrhea in heifers. Thirty-two newborn Holstein heifers were divided into two groups with sixteen animals each: the control group and the treated group that received sodium selenite (0.2 mg/kg) and vitamins A (35 mg/kg) and E (1 mg/kg) with one day of life, and a second application associated with secnidazole (400 mg/animal) on day 10 of life. Sample collection (blood and feces) were performed on days 1, 15, 30, 45 and 60 of life. Heifers from the treated group showed higher hematocrit values compared to the control group on day 60 of life, while total serum protein levels were higher on days 15 and 30. The ceruloplasmin (days 15, 30 and 60), IgG of heavy chain (days 15, 30, 45 and 60), IgG of light chain (days 45 and 60) and haptoglobin (days 15, 30, 45 and 60) were higher in the treated group compared to the control group. Serum levels of glucose decreased in treated animals on day 60 of life, while serum levels of albumin, triglycerides, urea, cholesterol, thiobarbituric acid reactive substances, reactive oxygen species and glutathione S-transferase activity did not differ between groups. Secnidazole was able to prevent infections caused by Giardia duodenalis in the first few days of life, but no difference was observed between groups. Moreover, there was no difference on total bacteria count and the incidence of diarrhea between groups. No difference on weight gain was observed on day 60 of life, but on day 210 of life treated animals had higher weight gain compared to the control group. Based on these evidences, we concluded that the injectable application of Se and vitamins (A and E) associated to secnidazole can improve the immunological system, and consequently, favor animals performance.

Homeopathic treatment as an alternative prophylactic to minimize bacterial infection and prevent neonatal diarrhea in calves

Bovine neonatal diarrhea is common due low immunity in newborn calves, poor management (or absence) of sanitary barriers, and other factors. Newborn calves with diarrhea in the first days of life suffer failure to thrive and may die if left untreated. The aim of this study was to evaluate whether prophylactic administration of a homeopathic product (Dia 100®) can control bovine neonatal diarrhea in calves born on a farm with substantial sanitary challenges. We counted total bacteria and protozoan parasites in fecal samples. We measured serum glucose, total protein, globulin, albumin, cholesterol and triglycerides on days 1, 7 and 14 of life. Twenty newborn calves were maintained in individual stalls, and were divided in two groups: ten untreated animals (control) and ten animals treated with Dia 100®. Fecal consistency was evaluated daily. We diagnosed diarrhea in five animals in the treated group, and in all animals from the control group. Infections with Escherichia coli and Giardia duodenalis were identified as the responsible organisms. The E. coli count was low in the treatment group on day 7 of life compared with the control group. Antibiotics were given to eight animals in the control group, and to two animals in the treatment group. On day of life 7, serum levels of total protein and globulins were higher in the control group, but were lower on day 14. Serum levels of glucose and triglycerides were greater in treated animals on days 7 and 14, suggesting that the homeopathic product contributes to improvement of intestinal health and absorption and nutrients. We conclude that Dia 100® controls diarrhea with 50% of efficacy, and reduces antibiotic utilization.

Ecotoxicology of Glycerol Monolaurate nanocapsules

Glycerol Monolaurate (GML) is a compound with known antimicrobial potential, however it is not much used due to its low solubility in water and high melting point. The nanoencapsulation of some drugs offers several advantages such as improved stability and solubility in water. The present study aimed to produce, characterize, and evaluate the ecotoxicity of GML nanocapsules. The nanocapsules were produced and presented a mean diameter of 210nm, polydispersity index of 0.044, and zeta potential of -23.4mV. The electron microscopy images showed the nanometric size and spherical shape. The assay in soil showed that GML has a high toxicity while the GML nanocapsules showed decreased toxic effects. Nanostructuration also protected the Rhamdia quelen against the toxic effects of GML. Concluding, the formulation shows positive results and is useful to predict the success of development besides not damaging the soil.

Risk factors for Toxoplasma gondii in sheep of southern Brazil

Toxoplasma gondii is responsible for reproductive failure in sheep, and it is considered a zoonotic parasite. Therefore, the aim of this study was to evaluate the prevalence of toxoplasmosis and its risk factors at the farm level. For that, 314 sheep from 18 farms of 7 municipalities of the West part of Santa Catarina State, Southern Brazil were used. To identify possible risk factors associated to the disease, a survey was applied to farmers with questions regarding farm characteristics, type of management, and history of animal reproductive problems. Out of 314 sera samples, 118 (37%, CI 95% 32.3–43.3) were positives for T. gondii. Multivariate analyses showed that allowing cats to have access to sheep feed was significantly associated to toxoplasmosis, with an odds ratio of 2.42. Similarly, keeping cats in the farm increased 1.67 times the chances of T. gondii infection by sheep. Therefore, we concluded that sheep infection by T. gondii in the West of Santa Catarina state is distributed very similarly to other regions of Brazil, and the presence of cats in the property is strongly correlated to the occurrence of the disease.