An evaluation of techniques to diagnose Dioctophyme renale in dogs.

Dioctophyme renale is a nematode with zoonotic potential that affects the kidneys of carnivorous, wild, and domestic mammals. In this study, we sought to evaluate the indirect ELISA method against routine methods used to diagnose dioctophimosis. Hence, 38 dogs parasitized by D. renale, as confirmed by surgery, were selected. The dogs were evaluated by abdominal ultrasound and urinalysis, and their sera were tested by indirect ELISA using D. renale adult secretion and excretion antigen (DES). Five dogs were followed up with serum collections on day 0 (day of surgery) and 30, 60, and 90 days after surgery to evaluate antibody kinetics. Abdominal ultrasound and indirect ELISA successfully diagnosed 37 dogs parasitized by D. renale, while urinalysis diagnosed 29 animals. The positive animals were parasitized with 1-7 parasites; 17 dogs were infected by male and female parasites, 15 only by female parasites, and six were parasitized only by male parasites. When assessing specificity and sensitivity, all techniques showed 100% specificity and 81.6%, 97.4%, and 97.4% sensitivity for urinalysis, ultrasound, and ELISA, respectively (p < 0.001). The five positive dogs that were followed up after surgery showed a progressive decrease in mean absorbances in indirect ELISA (0.644, 0.516, 0.511, and 0.440, respectively). This study demonstrated that the indirect ELISA using the DE antigen could diagnose dioctophimosis regardless of the number, sex, and location of the parasites, with the potential to be used in epidemiological research and implementing immunological and molecular studies, opening new lines of research on D. renale.

Dioctophyme renale é um nematódeo que possui potencial zoonótico e acomete o rim de mamíferos carnívoros, silvestres e domésticos. Neste estudo busca-se avaliar a técnica de ELISA indireto frente metodologias de rotina utilizadas no diagnóstico da dioctofimose. Para isto, 38 cães participaram do estudo, sendo todos parasitados por D. renale, confirmados por cirurgia. Esses cães foram avaliados por ultrassom abdominal, urinálise e seus soros testado por ELISA indireto utilizando antígeno de excreção e secreção (DES) de adultos de D. renale. Destes, cinco cães foram acompanhados com coletas de soro, no dia zero (dia da cirurgia) e após 30, 60 e 90 dias da cirurgia para avaliar a cinética dos anticorpos. O ultrassom abdominal e ELISA indireto apresentaram sucesso no diagnóstico de 37 cães parasitados por D. renale, enquanto que a urinálise diagnosticou 29 animais. Os animais positivos possuíam de um a sete parasitos, 17 cães apresentaram infecções por macho e fêmeas, 15 somente por fêmeas e seis foram parasitados apenas por machos. Na avaliação da especificidade e sensibilidade, todas as técnicas apresentaram 100% de especificidade e 81,6%, 97,4%, 97,4% de sensibilidade para urinálise, ultrassom e ELISA, respectivamente (p < 0,001). Os cinco cães positivos que foram acompanhados após a cirurgia apresentaram diminuição progressiva nas médias de absorbâncias no ELISA indireto (0,644, 0,516, 0,511 e 0,440, respectivamente). O estudo demonstrou que o ELISA indireto utilizando o antígeno DES poderia diagnosticar dioctofimose, independentemente do número, sexo e localização dos parasitos, com potencial para ser utilizada em estudos epidemiológicos e na implementação de estudos imunológicos e moleculares, abrindo novas linhas de pesquisa sobre Dioctophyme renale.

Activity of compounds derived from benzofuroxan in Trichomonasvaginalis.

Trichomoniasis is a sexually transmitted infection caused by the protozoan Trichomonas vaginalis. Currently, trichomoniasis is treated with the class of nitroimidazoles, namely, metronidazole; however, resistant isolates and strains have been reported. The compounds derived from benzofuroxan are biologically active heterocycles. This study evaluated the in vitro antiparasitic activity of these compounds in trophozoites of T. vaginalis and determined the mean inhibitory concentration (IC50), minimum inhibitory concentration (MIC), mortality curve, and cytotoxicity. The compounds were named EH1, EH2, EH3, and EA2 and tested in various concentrations: 100 to 15 µM (EH1 and EH2); 100 to 5 µM (EH3); and 100 to 25 µM (EA2), respectively. The greatest efficacy was observed in the highest concentrations in 24 h, with inhibition of approximately 100% of trophozoites. Compounds EH2 and EH3 had the lowest MIC: EH2 (35 µM) and EH3 (45 µM), with IC50 of 11.33 µM and 6.83 µM, respectively. Compound EA2 was effective at the highest concentrations. The activity of the compounds in T. vaginalis started in the first hour of incubation with 90% inhibition; after 12 h, inhibition >95% was observed. Compound EH1 showed the lowest activity, with the highest activity between 12 and 24 h after incubation. These results demonstrate that benzofuroxan derivatives are promising compounds for the in vitro treatment of T. vaginalis.

Antiparasitic treatment using herbs and spices: A review of the literature of the phytotherapy.

This study sought to make a literature review of the medicinal plants Origanum majorana, Origanum vulgare L., Thymus vulgaris L., Cuminum cynimum L., and Rosmarinus officinalis L. with antiparasitic potential. Articles and theses were selected from the LILACS, PubMed, and Google Scholar databases, which comprised the period from 2000 to 2021 (22 years). In all, 49 studies were selected, and the majority were with the plant Origanum vulgare L. (oregano), followed by Thymus vulgaris L. (thyme). Twenty-five genera of parasites were detected, which were described being tested with phytotherapic. The nematode Haemonchus spp. was the most evaluated in these studies, followed by the parasite genera Leishmania, Trichostrongylus, and Toxocara. All plants showed antiparasitic effects, with more or less action, therefore with the potential to continue research in the search for biomolecules to control these parasites.

O presente trabalho faz uma revisão bibliográfica das plantas medicinais Origanum majorana, Origanum vulgare L., Thymus vulgaris L., Cuminum cynimum L. e Rosmarinus officinalis L. com potencial antiparasitário. Foram selecionados artigos e teses nos bancos de dados LILACS, PubMed e Google Acadêmico que compreendiam o período de publicação de 2000 a 2021 (22 anos). Ao todo, foram selecionados 49 estudos, sendo que na maioria constava a planta Origanum vulgare L. (orégano), seguido de Thymus vulgaris L. (tomilho). Foram detectados 25 gêneros de parasitos, os quais foram descritos sendo testados frente a algum fitoterápico. O nematoda Haemonchus spp. foi o mais avaliado nestes estudos, seguido dos gêneros dos parasitos Leishmania, Trichostrongylus e Toxocara. Todas as plantas apresentaram efeitos antiparasitários, com maior ou menos ação, portanto com potencial para dar continuidade aos estudos em buscas de biomoléculas para controle destes parasitos.

Sheep immune-stimulated with Saccharomyces boulardii show reduced prolificacy of Haemonchus contortus.

Haemonchus contortus is the most pathogenic parasite for sheep. The objective was to evaluate immunomodulation of the probiotic Saccharomyces boulardii in sheep experimentally infected with H. contortus. Twenty-four sheep were divided into three groups: one infected with 500 H. contortus larvae/day for 26 days and supplemented with S. boulardii (40 ml with 1 × 108 CFU/ml/day); a control group only infected with H. contortus but not supplemented; and a naïve group that never came into contact with either parasites or S. boulardii. To assess the humoral immune response, production of specific serum IgG anti-somatic H. contortus antigen was evaluated through indirect ELISA. To assess the cellular immune response, cell populations and cytokine (IL-4, IL-5 and IL-10) production were evaluated through flow cytometry. For parasitological analyses, the counts of eggs per gram of faeces (EPG) and larvae per faecal culture were assessed. At all the study points, the concentration of IgG anti-H. contortus was higher (p < .05) in the S. boulardii group than in the other groups. The cell analysis revealed that there were significantly higher numbers (p < .05) of cells expressing MHC-II and significantly higher numbers (p < .05) of eosinophils in the mucosa in the S. boulardii group. Significant expression of IL-10 was observed only in the control infected group. There were significant reductions (p < .05) in EPG and larval counts in the S. boulardii supplemented group. These results show that S. boulardii supplementation modulated the immune response against H. contortus, thereby reducing its infection.

Ovicidal activity of the hydroalcoholic extract of Brazilian peppertree (Schinus terebinthifolia Raddi) against Ancylostoma spp. from naturally parasitized dogs.

This study aimed to evaluate the ovicidal activity of the hydroalcoholic extract of Schinus terebinthifolia (SCH; T1) against Ancylostoma spp. and its influence of storage time in the extract stored for 36 months (T36). Eggs of Ancylostoma spp. were obtained from naturally parasitized dogs, and used for the larval hatchability test, where the eggs were exposed to T1 and T36 extracts of SCH (15-0.625 mg/mL). In T1, all concentrations inhibited more than 80% of the eggs, being 100% at concentrations between 15 and 5 mg/mL (p > 0.05). At T36, all concentrations were active, even the ones between 2.5 and 0.625 mg/mL, with 100% inhibition (p < 0.05), revealing that the storage time maintained the ovicidal action. By LC-MS, T36 presented ethyl gallate, myricitrin, and gallic acid as major compounds. These findings support the promising use of SCH extract as an ovicide against Ancylostoma spp., even stored for 36 months of shelf life.

Quantification of Toxocara canis DNA by qPCR in mice inoculated with different infective doses.

The nematode Toxocara canis is of public health importance and is the main causative agent of toxocariasis in humans. This disease is difficult to diagnose due to several factors, including the possibility of cross-reactions with other nematodes in the ELISA. To overcome this problem, molecular tests have been recommended as an alternative to identify the parasite. The quantitative real-time polymerase chain reaction (qPCR) technique was used in this study to identify and quantify the parasite load of T. canis in the mouse brain. To this end, 24 mice were divided into six groups, five of which were challenged with different infective doses of T. canis larvae (L3) (1000, 500, 250, 100 and 50 larvae), while the sixth group, uninfected, acted as negative control. Forty-five days after infection, the animals were euthanized to collect the brain, from which two portions of 20 mg of tissue were taken for DNA extraction, while the rest of the brain tissue was digested to quantify the number of larvae by microscopy. The number of DNA copies was calculated from the standard DNA quantification curve, showing values of E = 93.4%, R2 = 0.9655 and Y = -3.415. A strong positive correlation (R = 0, 81; p < .001) was found between the number of copies and the recovery of larvae from brain. However, the parasite's DNA was also identified even in animals from whose brain no larvae were recovered after tissue digestion. The results of this study therefore confirm that the qPCR technique can be a valuable tool for the detection and quantification of T. canis DNA in murine hosts, even in animals whose with tissues contain very few parasites.

Detection of Toxocara canis DNA in tissues of experimentally infected mice.

The main etiological agent of toxocariasis is the helminth Toxocara canis. Several difficulties are found in the diagnosis of this disease, because of nonspecific clinical signs and possible cross-reactions that may occur in the available test, the indirect ELISA. Therefore, molecular diagnosis has been indicated as an alternative to conventional diagnosis. The purpose of this study was to evaluate the polymerase chain reaction (PCR) technique for the identification of T. canis in tissues of experimentally infected mice. To this end, nine mice were inoculated with 1500 embryonated eggs and were divided into two groups, the first euthanized 48 h (G1) and the other 30 days post inoculation (G2). Lungs, brain, liver and blood were collected from all the animals for DNA Extraction and tissue digestion, also was collected blood samples for DNA extraction and ELISA test (serum). Toxocara canis DNA was identified in all the inoculated animals using the ITS-2 target gene. The PCR test successfully identified the parasite in the brain, lung and liver of the animals euthanized 48 h PI and 30 days PI. This technique yielded good results in the identification of the parasite in the brain, being more sensitive than the method for the recovery of larvae, in the group with acute infection (48 h PI). The infection was confirmed by PCR within 48 h after infection, while the ELISA indicated serological conversion occurred only 14 days after inoculation. This study demonstrates the ability of PCR to identify T. canis in the liver, lungs and brain during acute and chronic infection.