Assessment of α-Cypermethrin Pour-On Application and Diminazene Aceturate for Treating Trypanosome-Related Diseases Caused by Tsetse Flies on Cattle in Mô, Togo

The effects of tsetse-transmitted trypanosomosis control in high tsetse flies (Glossina spp.) challenge and trypanocidal drug resistance settings remain poorly understood in Togo owing to poor data coverage on the current disease impact. From March 2014 to November 2017, a database of zoo-sanitary surveys integrating the evolution of disease incidence and intervention coverage made it possible to quantify the apparent effects attributable to the control effort, focused on all sedentary cattle breeds in the 1,000 km² area of Mô in Togo. The strategy involved an initial phase with cross-sectional entomological and parasitological. Then, three times a year, 20% of the bovine animals of the study area received α-cypermethrin pour-on, and infected cattle with poor health (798 cattle in 2014 and 358 in 2017) were individually given diminazene aceturate at 7 mg/kg of body weight. The tsetse density in the area decreased significantly, from 1.78 ± 0.37 in March 2014 before the α-cypermethrin application to 0.48 ± 0.07 in February 2017. The α-cypermethrin pour-on application and diminazene aceturate treatment of cattle led to the largest reduction in disease incidence, from 28.1% in 2014 to 7.8% in 2017, an improvement in hematocrit from 24.27 ± 4.9% to 27.5 ± 4.6%, and a reduction in calf mortality from 15.9 ± 11% to 5.9%. Improved access to these interventions for different types of livestock and maintaining their effectiveness, despite high tsetse (Diptera: Glossinidae) challenges, should be the primary focus of control strategies in many areas of Togo.

Discovering the Potent Inhibitors Against Babesia bovis in vitro and Babesia microti in vivo by Repurposing the Natural Product Compounds

In the present study, we screened 502 natural product compounds against the in vitro growth of Babesia (B.) bovis. Then, the novel and potent identified compounds were further evaluated for their in vitro efficacies using viability and cytotoxicity assays. The in vivo inhibitory effects of the selected compounds were evaluated using B. microti “rodent strain” in mice model. Three potent compounds, namely, Rottlerin (RL), Narasin (NR), Lasalocid acid (LA), exhibited the lowest IC50 (half-maximal inhibitory concentration) as follows: 5.45 ± 1.20 μM for RL, 1.86 ± 0.66 μM for NR, and 3.56 ± 1.41 μM for LA. The viability result revealed the ability of RL and LA to prevent the regrowth of treated parasite at 4 × IC50 and 2 × IC50, respectively, while 4 × IC50 of NR was sufficient to stop the regrowth of parasite. The hematology parameters of B. microti in vivo were different in the NR-treated groups as compared to the infected/untreated group. Interestingly, intraperitoneal administration of NR exhibiting inhibition in the growth of B. microti in mice was similar to that observed after administration of the commonly used antibabesial drug, diminazene aceturate (DA) (76.57% for DA, 74.73% for NR). Our findings indicate the richness of natural product compounds by novel potent antibabesial candidates, and the identified potent compounds, especially NR, might be used for the treatment of animal babesiosis.

Compounds from the Medicines for Malaria Venture Box Inhibit In Vitro Growth of Babesia divergens, a Blood-Borne Parasite of Veterinary and Zoonotic Importance

Babesiosis is an infectious disease with an empty drug pipeline. A search inside chemical libraries for novel potent antibabesial candidates may help fill such an empty drug pipeline. A total of 400 compounds (200 drug-like and 200 probe-like) from the Malaria Box were evaluated in the current study against the in vitro growth of Babesia divergens (B. divergens), a parasite of veterinary and zoonotic importance. Novel and more effective anti-B. divergens drugs than the traditionally used ones were identified. Seven compounds (four drug-like and three probe-like) revealed a highly inhibitory effect against the in vitro growth of B. divergens, with IC50s ≤ 10 nanomolar. Among these hits, MMV006913 exhibited an IC50 value of 1 nM IC50 and the highest selectivity index of 32,000. The atom pair fingerprint (APfp) analysis revealed that MMV006913 and MMV019124 showed maximum structural similarity (MSS) with atovaquone and diminazene aceturate (DA), and with DA and imidocarb dipropionate (ID), respectively. MMV665807 and MMV665850 showed MMS with each other and with ID. Of note, a high concentration (0.75 IC50) of MMV006913 caused additive inhibition of B. divergens growth when combined with DA at 0.75 or 0.50 IC50. The Medicines for Malaria Venture box is a treasure trove of anti-B. divergens candidates according to the obtained results.

ACE2/ANG-(1-7)/Mas Receptor Axis Activation Prevents Inflammation, Glial Activation And Cognitive Decline In a Rat Model of STZ Induced Impairment of Learning and Memory

Activation of the renin-angiotensin system (RAS), mediated by Angiotensin converting enzyme/Angiotensin II/Angiotensin receptor-1 (ACE/Ang II/AT1 R) axis elicits amyloid pathology, induces neurodegeneration and cognitive impairment leading to Alzheimer's disease (AD). On the contrary, Angiotensin converting enzyme2 (ACE2) produces Ang -(1-7) which binds with the Mas receptor and counters ACE/Ang II/AT1 axis. To date, the involvement of ACE2/Ang-(1–7)/MasR axis in etiology and progression of AD largely remains to be elucidated. Hence, the present study is aimed to determine the role of ACE2/Ang-(1–7)/MasR axis in STZ induced model of neurodegeneration using Diminazene aceturate (DIZE), an ACE2 activator in both in vitro/in vivo experimental conditions. Interestingly, ROS content and oxidative stress burden in N2A cells were found to be attenuated along with a decrease in enzymatic activity of AChE following DIZE treatment. In contrast, activation of this axis led to altered mitochondrial membrane potential (MMP) in addition to ablated intracellular Ca2+ influx. ACE2/Ang-(1–7)/MasR axis activation further resulted in reduction of astrogliosis as indicated by decreased intensity of NFκB and dwindled expression of its downstream NLRP3 cascade signaling molecules. These results were confirmed by using a selective inhibitor of ACE-2, MLN-4760, which reversed the protective effects of ACE2 activation by DIZE. Subsequently, treatment with DIZE in STZ induced rat model of AD prevented cognitive impairment and progression of amyloid pathology. Therefore, the involvement of ACE2/Ang-(1–7)/Mas axis suggests that it could be further explored as a potential pathway in AD, owing to its inhibitory effect on inflammation/astrogliosis and restoring cognitive functions.

Effects of Methanolic Extract from Turmeric (Curcuma longa) against the In Vitro Multiplication of Several Babesia Species and Theileria equi

Anti-piroplasm drugs currently on the market have proven toxicity to the host and parasite resistance. Plants are possible sources of novel drugs. Subsequently, a novel strategy should be used to find new anti-piroplasm agents that are both effective and safe. In the present study, we have evaluated the effect of turmeric (Curcuma longa) methanolic extract on the in vitro growth of Babesia (B.) bovis, B. divergens, B. caballi, and Theileria (T.) equi. The in vitro inhibitory effectiveness of turmeric was assessed using a fluorescence test. The enhancement in the in vitro inhibitory efficacy of turmeric when administrated in combination with diminazene aceturate (DA) was investigated using in vitro cultures of different piroplasm parasites. Turmeric reduced the in vitro growth of B. bovis, B. divergens, T. equi, and B. caballi with IC50 values of 0.830 0.078, 0.375 0.055, 1.405 0.575, and 0.720 0.090 mg/mL, respectively. An amount of 1 mg/mL turmeric for B. bovis, 0.5 mg/mL turmeric for B. divergens, 1 mg/mL turmeric for T. equi, and 0.5 mg/mL turmeric for B. caballi exhibited 73.43%, 80.065%, 73.47%, and 47.375% inhibitions in the growth of the parasites, respectively. When turmeric was combined with DA, its in vitro inhibitory impact on bovine Babesia and equine Babesia/Theileria parasites was amplified. These findings show that a methanolic extract of turmeric could be a promising medicinal plant for the treatment of babesiosis, especially when administered in conjunction with DA.

First Molecular Study of Caprine Trypanosoma evansi Infection in Central India

Background: Diagnosis of Trypanosoma evansi among goat in field is still challenging due to scarce information on clinical manifestation and non availability of rapid reliable diagnostics. Molecular test such as PCR employed to diagnose the T. evansi with accuracy in both blood and tissues. Limited reports are available from different parts of the country on natural infection of T. evansi in goat. Mainly subclinical form of disease occurred in goats owing to low level of parasitaemia of T. evansi. Methods: A Jamunapari goat aged 1.5 years and weighing ~25 Kg was suffering from circling movement, pale conjunctiva and anorexia and was found positive for T. evansi by buffy coat examination and polymerase chain reaction. Diminazene aceturate was given intramuscularly as 7% water solution at a dose of 3.5 mg/kg b.wt. After 3 days, blood smear and buffy coat were negative for T. evansi and no band was found after running the product on 1.7% agarose gel stained with ethidium bromide. Result: The present study reveals first time report on caprine trypanosomosis from central India by conventional and PCR technique and placed on record. Hence, the goat should also be taken into consideration for control of trypanosomosis as goats are an important source of infection to other animals due to their reservoir nature for T. evansi.

Evaluasi Pemberian Obat Diminazene Aceturate Secara In Vivo Pada Mencit (Mus musculus) yang Diinfeksi Isolat Trypanosoma evansi

Surra's disease is caused by Trypanosoma evansi parasite has been established as one of the strategic infectious animal diseases. Drug resistance in this case is one of the major challenges in handle and control them. The aim of this study is to evaluate the provision drug resistance diminazene aceturate (Tryponil®) on Trypanosoma evansi isolate from Pemalang and Brebes Central Java province with in vivo test in mice. Total 50 mice, BALB / c strain, male, 2 months, body weight ± 30 gram are obtained from LPPT-UGM, adapted for one week. Mice were divided into 10 groups consist of 5 each. Each mouse was infected with Trypanosoma evansi by intraperitonial route. Treatment was given when mice had reached the level of parasitemia 108 – 109 trypanosoma / mL of blood this was predicted 24 hours post-infection (Eisler et al., 2001). The administration of the drug tryapanosidal was done intraperitonial with doses 1mg/kg, 3mg / kg, 5 mg / kg and 7mg / kg. Observation of parasitemia did every 2 times in one week till 60 days of observation. Parasitemia observation was performed using 3 techniques. The first method was native examination used a microscope, if the negative results would be followed by MHCT (Microhaematocrit centrifugation Technique) and BCT (Buffy Coat Technique) according to OIE (2012). Data obtained from the treatment group were the level of parasitemia, the number of deaths and the number of live mice from each test dose. The results are analysed by standard logit or probit. The results of this study showed the effects of the drug Dimianzene aceturate on both isolates varied. On Brebes Isolate was effective at doses of 7 mg / kg BW (100%) and 5mg / kg BW (80%), whereas in the effective dose Pemalang isolate at 3 mg dose / kg BW (80%), 5 and 7 mg / kg BW (100%). While at the lowest dose of 1 mg / kg obtained a level of effectiveness of 0% in both isolates. It could be concluded that both isolates have different pathogens and indicate resistance subpopulation to diminazene aceturate.Keywords : diminazene aceturate, in vivo, resistance, Trypanosoma evansi.

Experimental Babesia rossi infection induces hemolytic, metabolic, and viral response pathways in the canine host

Background Babesia rossi is a leading cause of morbidity and mortality among the canine population of sub-Saharan Africa, but pathogenesis remains poorly understood. Previous studies of B. rossi infection were derived from clinical cases, in which neither the onset of infection nor the infectious inoculum was known. Here, we performed controlled B. rossi inoculations in canines and evaluated disease progression through clinical tests and whole blood transcriptomic profiling. Results Two subjects were administered a low inoculum (104 parasites) while three received a high (108 parasites). Subjects were monitored for 8 consecutive days; anti-parasite treatment with diminazene aceturate was administered on day 4. Blood was drawn prior to inoculation as well as every experimental day for assessment of clinical parameters and transcriptomic profiles. The model recapitulated natural disease manifestations including anemia, acidosis, inflammation and behavioral changes. Rate of disease onset and clinical severity were proportional to the inoculum. To analyze the temporal dynamics of the transcriptomic host response, we sequenced mRNA extracted from whole blood drawn on days 0, 1, 3, 4, 6, and 8. Differential gene expression, hierarchical clustering, and pathway enrichment analyses identified genes and pathways involved in response to hemolysis, metabolic changes, and several arms of the immune response including innate immunity, adaptive immunity, and response to viral infection. Conclusions This work comprehensively characterizes the clinical and transcriptomic progression of B. rossi infection in canines, thus establishing a large mammalian model of severe hemoprotozoal disease to facilitate the study of host-parasite biology and in which to test novel anti-disease therapeutics. The knowledge gained from the study of B. rossi in canines will not only improve our understanding of this emerging infectious disease threat in domestic dogs, but also provide insight into the pathobiology of human diseases caused by Babesia and Plasmodium species.