Efficacy and safety of geranium-oregano-thymol formulations to control of dog tick Rhipicephalus sanguineus sensu lato under laboratory and field conditions.

The present study evaluated, in laboratory and field, the efficacy and safety of formulations of Pelargonium graveolens (geranium - G), Origanum majorana (oregano - O) commercial essential oils (EO) and thymol (T) to control of Rhipicephalus sanguineus sensu lato. In the laboratory, three formulas (A: 2% tween 80%, B: powder and C: nanoemulsion) by a mixture of these components (GOT) were prepared and evaluated, and the best one was used to assess its safety and field application against R. sanguineus s. l. on naturally infested dogs. Besides the major compounds of the EO used were identified. The results of the lab study showed that formula A (2.5 g of each G + O + T + 2% tween 80 to complete 100 mL) was significantly more effective than the other two formulas tested and exhibited highly effective adulticidal, larvicidal, and ovicidal activity against R. sanguineus s.l. Significant LC50 and LC90 values of GOT were evaluated (13.4 and 21.5 mg/mL, respectively) for the adulticidal activity, (2.81 and 4.46 mg/mL, respectively) for ovicidal activity and (2.44 and 4.45 mg/mL, respectively) for larvicidal activity. The safety of formula A has been proven by the absence of its cytotoxicity on a cell line of human epidermoid carcinoma. Citronella and carvacrol were the major compounds identified in the commercial essential oils of P. graveolens and O. majorana, respectively. Formula A was used in a field control trial for almost 8 months, during the tick infestation season (April to November, 2022). Fourteen naturally infested dogs were divided into two groups, each with seven dogs. One group received formula A spraying five times during an experiment that continued for 8 months, while the other group received treatment with commercially available malathion acaricide. The animals were sprayed on five occasions throughout the experiment (April, June, July, August, and September). The results showed a substantial percentage of effectiveness after the first application of formula A with a 99.3% reduction in tick count at day 28 post-application (PA). In the case of severe infestation 60 days after the first application of formula A (more than 180 ticks per dog), the second application was done, achieving an efficacy of 54.9% at day 3 PA, so an emergency spray was done at day 5 PA to combat the rest of the tick infestation, achieving efficacy of 99% after 3 days. Consequently, a regular spray (third, fourth, and fifth application) was done every 35 days. This regular spray revealed 100% effectiveness at 14 days PA. Biochemical parameters of treated dogs were evaluated to confirm the safety of formula A. Creatinine, ALT, and albumin of the dogs treated with formula A were within the normal range of dogs, while urea and AST were higher than the normal range. In conclusion, formula A can safely treat R. sanguineus s.l. infestations in dogs with regular application every 5 weeks.

An investigation of the acaricidal activity of benzyl alcohol on Rhipicephalus annulatus and Rhipicephalus sanguineus and its synergistic or antagonistic interaction with commonly used acaricides.

The most economically significant ectoparasites in the tropics and subtropics are ixodid ticks, especially Rhipicephalus annulatus and Rhipicephalus sanguineus. Years of extensive use of the readily available acaricides have resulted in widespread resistance development in these ticks, as well as negative environmental consequences. Benzyl alcohol (BA) has been frequently used to treat pediculosis and scabies, and it may be an effective alternative to commonly used acaricides. The main aim of the present study was to evaluate the acaricide activity of BA and its combination with the regularly used chemical acaricides against R. annulatus and R. sanguineus. Different concentrations of BA alone and in combination with deltamethrin, cypermethrin and chlorpyrifos were tested in vitro against adult and larvae of both tick species. The results showed that BA is toxic to R. annulatus and R. sanguineus larvae, with 100% larval mortality at concentrations of ≥50 mL/L, and LC50 and LC90 attained the concentrations of 19.8 and 33.8 mL/L for R. annulatus and 18.8 and 31.8 mL/L for R. sanguineus, respectively. Furthermore, BA in combination with deltamethrin, cypermethrin and chlorpyrifos exhibited synergistic factors of 2.48, 1.26 and 1.68 against R. annulatus larvae and 1.64, 11.1 and 1.14 against R. sanguineus larvae for deltamethrin + BA, cypermethrin + BA and chlorpyrifos + BA, respectively. BA induced 100% mortality in adult R. annulatus at concentrations of ≥250 mL/L with LC50 and LC90 reached the concentrations of 111 and 154 mL/L, respectively. Additionally, BA had ovicidal activity causing complete inhibition of larval hatching at 100 mL/L. The combination of BA with deltamethrin and cypermethrin increased acetylcholinesterase inhibition, whereas the combination of BA with chlorpyrifos decreased glutathione (GSH) activity and malondialdehyde levels. In the field application, the combination of BA 50 mL/L and deltamethrin (DBA) resulted in a significant reduction in the percentage of ticks by 30.9% 28 days post-treatment when compared with groups treated with deltamethrin alone. In conclusion, BA causes mortality in laboratory and field studies alone and in combination with cypermethrin or deltamethrin. BA can be used for control of ticks of different life stages, that is, eggs and larvae, through application to the ground.

Effects of iron chelating agent on Schistosoma mansoni infected murine model.

Schistosomiasis is one of the major health problems in many tropical and developing countries. Infection takes place once cerceriae penetrate human skin, then it changed into schistosomules. The schistosomules takes iron in the form of heme from host's haemoglobin, ferritin and transferrin. Iron is a vital element not only for growth and sexual maturity of schistosomules to adults but also for oogenesis. Since the trapped eggs are the pathological causative agent for most of pathogenesis and complications, the current work was designed to study the effects of early deprivation of schistosomules from iron in the host (in vivo) by chelating it with deferoxamine (DFO). The iron chelation has effects on growth, maturity and egg deposition, as well as it has ameliorative effects on liver pathology such as hepatic fibrosis. Mice were classified into four groups, normal control, DFO treated only, Schistosoma mansoni (S. mansoni) infected DFO untreated and S. mansoni infected DFO treated. The infected DFO treated mice showed significant reduction in fecal egg excretion with increased percentage of dead eggs and this was accompanied with a significant reduction of both total worm burden and hepatic egg load and increased dead egg percentage compared to the infected DFO untreated group. There was also a significant reduction in both serum and hepatic tissue ferritin concentrations in the infected DFO treated mice in comparison to the infected DFO untreated group. Additionally, a significant decrease in number and size of granulomas with subsequent improvement of liver fibrosis was recorded in the infected DFO treated group. This immunopathology was also associated with significant up regulation of Interlukine12 (IL12), Interferon gamma (IFN γ) and significant down regulation in interleukin4 (IL4), interleukin10 (IL10) in both serum and hepatic tissue in the infected DFO treated compared to other groups. Entirely, DFO succeeded in diminishing the growth, maturity and fecundity of S. mansoni with a subsequent improvement of hepatic pathology. As a result of the above findings, it can be concluded that DFO could be considered as a useful treatment against schistosomal infection.

Immunopathological assessments of human Blastocystis spp. in experimentally infected immunocompetent and immunosuppresed mice.

Blastocystis spp., one of the most common parasites colonizing the human intestine, is an extracellular, luminal protozoan with controversial pathogenesis. The host's immune response against Blastocystis spp. infection has also not been defined yet. Therefore, this research aimed to assess the potential pathogenicity of this parasite and its ability to modulate the immune response in experimental infected immunocompetent and immunosuppresed mice. These results demonstrated that the infected immunosuppressed mice were more affected than infected immunocompetent mice. Histopathological examination of the small intestine in the infected immunosuppressed mice showed that Blastocystis spp. infiltrated all the layers. Moreover, the epithelia showed exfoliation and inflammatory cell infiltration in submucosa compared to that of the infected immunocompetent mice. As well, examination of the large intestine of the infected immunosuppressed group showed severe goblet cell hyperplasia. Blastocystis spp. infiltrated all the large intestine layers compared to that of the infected immunocompetent group. Furthermore, there was a significant upregulation of the expression of proinflammatory cytokines: interleukin 12 (IL-12) and tumor necrosis factor alpha (TNF-α) in the infected immunosuppressed mice compared to that of the infected immunocompetent ones (p ≤ 0.004 and p ≤ 0.002, respectively). However, the expression of anti-inflammatory cytokines (IL-4 and IL-10) was significantly downregulated in the infected immunosuppressed group compared to that of the infected immunocompetent group one at 10 days postinfection (p ≤ 0.002 and p ≤ 0.001, respectively). The results of this study revealed that Blastocystis spp. affected the production of pro- and anti-inflammatory cytokines in both groups of mice compared to healthy normal (naive) group. Additionally, these data showed that there was a significant upregulation (p ≤ 0.005) of the locally synthesized antibody: secretary IgA (sIgA) in the gut of the infected immunocompetent mice when compared to that of the infected immunosuppressed ones.

Quantitative assessment of the effects of sulfamethoxazole on Toxoplasma gondii loads in susceptible WT C57BL/6 mice as an immunocompetent host model.

The abundance of Toxoplasma gondii with or without sulfamethoxazole (SMX) treatment was evaluated with quantitative competitive polymerase chain reaction in various organs of wild-type C57BL/6 mice, a susceptible immunocompetent host, after peroral infection with a cyst-forming Fukaya strain of T. gondii. SMX affected different organs in three ways: T. gondii was reduced independently of SMX (skin and kidney); T. gondii was not eradicated with continuous treatment (brain, heart, and lung); and T. gondii was eradicated with continuous treatment (tongue, skeletal muscle, and small intestine). The SMX concentrations in the brains, hearts, and lungs were higher in infected mice than in uninfected mice. These results indicate that even in an immunocompetent host, chemotherapy is necessary to reduce the parasite load and thus reduce the risk of recurrent disease.

In vivo antiprotozoan effects of garlic (Allium sativum) and ginger (Zingiber officinale) extracts on experimentally infected mice with Blastocystis spp.

Controversy surrounding the pathogenic role of Blastocystis spp. in humans and lack of well-established diagnostic criteria led to debates concerning the treatment for that organism. Furthermore, some strains develop resistance against the recommended drugs. Thus, using natural medicine has many positive aspects to address these points. In an earlier study, we addressed in vitro effect of garlic and ginger on Blastocystis spp. isolates as an alternative treatment. Accordingly, this study was conducted to evaluate in vivo activities of these two herbs on mice infected with Blastocystis spp. Antiprotozoan activities were determined by monitoring Blastocystis shedding in stools and histopathological changes of the intestine of infected mice. Additionally, assessment of the antioxidant effect (via measuring the level of malondialdehyde (MDA) production) of these herbs on the treated groups of mice was done. Also, their effects on nitric oxide (NO) production were assessed. In this work, treatment of infected mice with garlic, ginger, and nitazoxanide (NTZ) reduced the shedding of cysts significantly compared to the infected untreated group, P value ≤0.001, 0.0001, and 0.0003, respectively. As well, histopathological examination revealed that Blastocystis was frequently observed within the lumen, at the tip of the epithelium, and/ or infiltrated in an enterocyte in the infected group without treatment compared to that of the infected treated ones. Furthermore, mice infected with Blastocystis exhibited increased levels of NO (440.09 ± 3.7 vs. 276.66 ± 0.8, P ≤ 0.001) and MDA production (106.19 ± 0.43 vs. 63.06 ± 0.45, P ≤ 0.0004) compared to that of the uninfected controls. Treatment of infected mice with garlic, ginger, and NTZ reduced NO levels to 54.41 ± 1.2, 47.70 ± 1.2, and 37.43 ± 0.98 and MDA levels to 22.38 ± 0.17, 63.34 ± 3.89, and 66.76 ± 9.1, respectively. We conclude that using ginger and garlic for treatment of blastocystosis is beneficial.

Lipocalin 2 bolsters innate and adaptive immune responses to blood-stage malaria infection by reinforcing host iron metabolism.

Plasmodium parasites multiply within host erythrocytes, which contain high levels of iron, and parasite egress from these cells results in iron release and host anemia. Although Plasmodium requires host iron for replication, how host iron homeostasis and responses to these fluxes affect Plasmodium infection are incompletely understood. We determined that Lipocalin 2 (Lcn2), a host protein that sequesters iron, is abundantly secreted during human (P. vivax) and mouse (P. yoeliiNL) blood-stage malaria infections and is essential to control P. yoeliiNL parasitemia, anemia, and host survival. During infection, Lcn2 bolsters both host macrophage function and granulocyte recruitment and limits reticulocytosis, or the expansion of immature erythrocytes, which are the preferred target cell of P. yoeliiNL. Additionally, a chronic iron imbalance due to Lcn2 deficiency results in impaired adaptive immune responses against Plasmodium parasites. Thus, Lcn2 exerts antiparasitic effects by maintaining iron homeostasis and promoting innate and adaptive immune responses.