Evaluation of the inhibitory effect of N-acetyl-L-cysteine on Babesia and Theileria parasites.

N-acetyl-L-cysteine is known to have antibacterial, antiviral, antimalarial, and antioxidant activities. Therefore, the in vitro inhibitory effect of this hit was evaluated in the present study on the growth of Babesia and Theileria parasites. The in vitro growth of Babesia bovis, Babesia bigemina, Babesia divergens, Theileria equi, and Babesia caballi that were tested was significantly inhibited (P < 0.05) by micromolar concentrations of N-acetyl-L-cysteine. The inhibitory effect of N-acetyl-L-cysteine was synergistically potentiated when used in combination with diminazene aceturate on B. bovis and B. caballi cultures. These results indicate that N-acetyl-L-cysteine might be used as a drug for the treatment of babesiosis, especially when used in combination with diminazene aceturate.

Clofazimine Inhibits the Growth of Babesia and Theileria Parasites In Vitro and In Vivo.

The present study evaluated the growth-inhibitory effects of clofazimine, currently used for treating leprosy, against Babesia bovis, B. bigemina, B. caballi, and Theileria equi in in vitro culture and against Babesia microti in mice. The 50% inhibitory concentrations (IC50s) of clofazimine against the in vitro growth of B. bovis, B. bigemina, B. caballi, and T. equi were 4.5, 3, 4.3, and 0.29 µM, respectively. In mice infected with B. microti, treatment with 20 mg/kg of body weight of clofazimine administered orally resulted in a significantly lower peak parasitemia (5.3%) than that in the control group (45.9%), which was comparable to the subcutaneous administration of 25 mg/kg diminazene aceturate, the most widely used treatment for animal piroplasmosis. Although slight anemia was observed in both clofazimine- and diminazene aceturate-treated infected mice, the level and duration of anemia were lower and shorter, respectively, than those in untreated infected mice. Using blood transfusions and PCR, we also examined whether clofazimine completely killed B. microti On day 40 postinfection, when blood analysis was performed, parasites were not found in blood smears; however, the DNA of B. microti was detected in the blood of clofazimine-treated animals and in several tissues of clofazimine- and diminazene aceturate-treated mice by PCR. The growth of parasites was observed in mice after blood transfusions from clofazimine-treated mice. In conclusion, clofazimine showed excellent inhibitory effects against Babesia and Theileria in vitro and in vivo, and further study on clofazimine is required for the future development of a novel chemotherapy with high efficacy and safety against animal piroplasmosis and, possibly, human babesiosis.

Identification and characterization of profilin antigen among Babesia species as a common vaccine candidate against babesiosis.

We have characterized a member of the profilin (PROF) family protein as a common antigen in three pathogens-Babesia bovis (B. bovis), Babesia bigemina (B. bigemina), and Babesia microti (B. microti)-and evaluated its immunogenic and cross-protective properties against a challenge infection with B. microti in BALB/c mice. The recombinant PROF proteins of B. bovis, B. bigemina, and B. microti were successfully expressed in Escherichia coli (E. coli) as soluble GST fusion proteins (rBboPROF, rBbigPROF, and rBmPROF, respectively), and they were found to be antigenic. On probing with mouse anti-rPROF serum, green fluorescence was observed on the parasites' cytosols by confocal laser microscopy. Immunization regimes in BALB/c mice using rPROFs induced cross-protective immunity against B. microti infection based on high levels of cytokines and immunoglobulin (IgG) titers, a reduction in peak parasitemia levels, and earlier clearance of the parasite as compared with control mice. The findings of the present study indicate that PROF is a common antigen among bovine and murine Babesia parasites, and it might be used as a common vaccine candidate against babesiosis.

Trichomonas gallinae: Prevalence and molecular characterization from pigeons in Minoufiya governorate, Egypt.

Trichomonas gallinae infects the upper digestive tract of pigeons. It is transmitted from mother to young squabs by feeding crop milk. Generally, infection resulted in severe mortalities in young birds. In this study, we examined 3315 pigeons of different ages from the Minoufiya governorate for the clinical infection by T. gallinae. The infection was confirmed in infected birds by microscopical examination of oral swabs, histopathological examination, and PCR of the ITS1/5.8S/ITS2 gene. The prevalence was 63 (1.9%). The parasite was found in 35 (2.04%) from Ashmoun, 15 (1.66%) from Minoof, 8 (1.6%) from Quesna, and 5 (2.5%) from El-Shohada birds. The infection was mainly detected in squabs 60 (1.8%). The sequence of T. gallinae ITS1/5.8S/ITS2 gene from Egypt has high nucleotide sequence identity (up to100%) to T. gallinae from pigeon of USA, Austria, Canada, and Spain. The sequence belongs to genotype B of T. gallinae. Histopathological examination presented the parasites in crop, liver, larynx, and trachea as poorly eosinophilic bodies with severe inflammatory cell infiltration. This is the first study to present the prevalence and genotype of T. gallinae from Minoufiya governorate, Egypt.

Evaluation of in vitro inhibitory effect of enoxacin on Babesia and Theileria parasites.

Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P < 0.05) by micro molar concentrations of enoxacin (IC50 values = 33.5, 15.2, 7.5 and 23.2 µM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or incase of animal sensitivity against diminazin toxicity.

Inhibitory effect of allicin on the growth of Babesia and Theileria equi parasites.

Allicin is an active ingredient of garlic that has antibacterial, antifungal, antiviral, and antiprotozoal activity. However, the inhibitory effects of allicin on Babesia parasites have not yet been examined. In the present study, allicin was tested as a potent inhibitor against the in vitro growth of bovine and equine Babesia parasites and the in vivo growth of Babesia microti in a mouse model. The in vitro growth of Babesia bovis, Babesia bigemina, Babesia caballi, or Theileria equi was inhibited by allicin in a dose-dependent manner and had IC50 values of 818, 675, 470, and 742 µM, respectively. Moreover, allicin significantly inhibited (P < 0.001) invasion of B. bovis, B. bigemina, B. caballi, and T. equi into the host erythrocyte. Furthermore, mice treated with 30 mg/kg of allicin for 5 days significantly (P < 0.05) reduced the parasitemia of B. microti over the period of the study. To further examine the potential synergism of allicin with diminazene aceturate, growth inhibitory assays were performed in vitro and in vivo. Interestingly, combinations of diminazene aceturate with allicin synergistically potentiated its inhibitory effects in vitro and in vivo. These results indicate that allicin might be beneficial for the treatment of babesiosis, particularly when used in combination with diminazene aceturate.

Prevalence and molecular characterization of Hysterothylacium species infecting Pandora (Pagellus erythrinus) in the Mediterranean Sea of Egypt.

Species of the genus Hysterothylacium are aquatic roundworms (nematodes) belonging to the family Raphidascarididae. Some species in this family are known to be associated with zoonotic diseases in humans after they consume their parasitic larvae in raw or undercooked fish. The aim of this research was to report the prevalence, morphology, and molecular characteristics of Hysterothylacium species in Pagellus erythrinus. A total of Two hundred fish were purchased from the fish market in Damanhour, Beheira Province, between December 2021 and November 2022 and subjected to examination. For molecular characterization, the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the mitochondrial cytochrome oxidase subunit 2 (COX-2) gene were used. Hysterothylacium species were morphologically described and identified from the intestine of Pagellus erythrinus in Beheira Province, Egypt. The PCR amplified 1087 bp and 629 bp of the target sequences of the ITS region and COX-2 gene, respectively. Sequence analysis revealed the Hysterothylacium thalassini species. The identified species provided novel biological data for the Hysterothylacium nematode in Pagellus erythrinus. The prevalence of Hysterothylacium species recovered from the intestine was 55%. The highest prevalence of 72% has been reported in summer compared to the lowest prevalence of 38% in the winter. Females had a higher prevalence of 61.8% than males, with 44.2%. The first detection, prevalence, and molecular characterization of H. thalassini in Pagellus erythrinus from Beheira Province, Egypt, was presented in this study.

Specific antibody to a conserved region of Babesia apical membrane antigen-1 inhibited the invasion of B. bovis into the erythrocyte.

Apical membrane antigen-1 (AMA-1) is a microneme protein that exists in all apicomplexan parasites and plays an indispensable role in the invasion into host cell. Central region of ectodomains I and II of Babesia bovis apical membrane antigen-1 (BbAMA-1P) is highly conserved with these of Babesia species and may be beneficial for vaccine development against babesiosis. In the present study, recombinant protein encoding the central region of B. bovis AMA-1 (rBbAMA-1P) was produced in Escherichia coli and its antiserum was prepared in mice for further molecular characterization. Anti-rBbAMA-1P serum specifically reacted with corresponding authentic protein of B. bovis as determined by Western blotting and IFAT. Cultured B. bovis treated with anti-rBbAMA-1P serum showed significant reduction in the in vitro growth of the parasites. Moreover, preincubated free merozoites with 1mg/ml anti-rBbAMA-1P serum inhibited their efficiency in the invasion into erythrocytes (RBCs) by 61% and 70% at 3h and 6h, respectively. Our data suggest that the central region of domains I and II of BbAMA-1 may serve as a vaccine candidate against babesiosis.

Molecular characterization and antigenic properties of a novel Babesia gibsoni glutamic acid-rich protein (BgGARP).

Identification and molecular characterization of Babesia gibsoni proteins with potential antigenic properties are crucial for the development and validation of the serodiagnostic method. In this study, we isolated a cDNA clone encoding a novel B. gibsoni 76-kDa protein by immunoscreening of the parasite cDNA library. Computer analysis revealed that the protein presents a glutamic acid-rich region in the C-terminal. Therefore, the protein was designated as B. gibsoni glutamic acid-rich protein (BgGARP). A BLASTp analysis of a translated BgGARP polypeptide demonstrated that the peptide shared a significant homology with a 200-kDa protein of Babesia bigemina and Babesia bovis. A truncated BgGARP cDNA (BgGARPt) encoding a predicted 13-kDa peptide was expressed in Escherichia coli (E. coli), and mouse antisera against the recombinant protein were used to characterize a corresponding native protein. The antiserum against recombinant BgGARPt (rBgGARPt) recognized a 140-kDa protein in the lysate of infected erythrocytes, which was detectable in the cytoplasm of the parasites by confocal microscopic observation. In addition, the specificity and sensitivity of enzyme-linked immunosorbent assay (ELISA) with rBgGARPt were evaluated using B. gibsoni-infected dog sera and specific pathogen-free (SPF) dog sera. Moreover, 107 serum samples from dogs clinically diagnosed with babesiosis were examined using ELISA with rBgGARPt. The results showed that 86 (80.4%) samples were positive by rBgGARPt-ELISA, which was comparable to IFAT and PCR as reference test. Taken together, these results demonstrate that BgGARP is a suitable serodiagnostic antigen for detecting antibodies against B. gibsoni in dogs.

Apicoplast-targeting antibacterials inhibit the growth of Babesia parasites.

The apicoplast housekeeping machinery, specifically apicoplast DNA replication, transcription, and translation, was targeted by ciprofloxacin, thiostrepton, and rifampin, respectively, in the in vitro cultures of four Babesia species. Furthermore, the in vivo effect of thiostrepton on the growth cycle of Babesia microti in BALB/c mice was evaluated. The drugs caused significant inhibition of growth from an initial parasitemia of 1% for Babesia bovis, with 50% inhibitory concentrations (IC(50)s) of 8.3, 11.5, 12, and 126.6 µM for ciprofloxacin, thiostrepton, rifampin, and clindamycin, respectively. The IC(50)s for the inhibition of Babesia bigemina growth were 15.8 µM for ciprofloxacin, 8.2 µM for thiostrepton, 8.3 µM for rifampin, and 206 µM for clindamycin. The IC(50)s for Babesia caballi were 2.7 µM for ciprofloxacin, 2.7 µM for thiostrepton, 4.7 µM for rifampin, and 4.7 µM for clindamycin. The IC(50)s for the inhibition of Babesia equi growth were 2.5 µM for ciprofloxacin, 6.4 µM for thiostrepton, 4.1 µM for rifampin, and 27.2 µM for clindamycin. Furthermore, an inhibitory effect was revealed for cultures with an initial parasitemia of either 10 or 7% for Babesia bovis or Babesia bigemina, respectively. The three inhibitors caused immediate death of Babesia bovis and Babesia equi. The inhibitory effects of ciprofloxacin, thiostrepton, and rifampin were confirmed by reverse transcription-PCR. Thiostrepton at a dose of 500 mg/kg of body weight resulted in 77.5% inhibition of Babesia microti growth in BALB/c mice. These results implicate the apicoplast as a potential chemotherapeutic target for babesiosis.

The use of different diagnostic tools for Babesia and Theileria parasites in cattle in Menofia, Egypt.

Bovine piroplasmosis is caused by tick-borne hemoprotozoans of the genera Babesia and Theileria and is the most prevalent in tropical and subtropical countries, causing a major economic impact worldwide. In the current study, a total of 405 cattle of different ages, sexes, and breeds were randomly sampled for surveying and diagnosis of babesiosis and theileriosis using three methods: direct microscopy (blood smears), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Giemsa-stained blood smears revealed that, out of 405 examined cattle, 33 (8.15 %) were infected with Babesia sp. and 65 (16.05 %) with Theileria sp. (total number of infected cattle was 98). Mixed infection was seen in 11 (2.72 %) animals. Moreover, application of the three diagnostic assays on 158 randomly sampled cattle indicated that 17 (10.76 %) and 33 (20.89 %) were positive for Babesia and Theileria spp. by the direct smear technique, 25 (15.82 %) and 33 (20.89 %) by IFAT (fluorescence was greenish yellow for Babesia and yellowish for Theileria), and 20 (12.66 %) and 38 (24.05 %) by PCR. Using primers specific for Babesia and Theileria spp., we found that diagnostic bands appeared at ~350 and ~370 bp, respectively indicating the presence of these piroplasms. Statistically, there was a non-significant difference of the positivity in response to the three techniques; thus, any of these methods can be described as useful for diagnosing blood parasites in both domesticated animals and birds. On the basis of the obtained results, it could be concluded that direct microscopy can be used in acute infections, whereas IFAT and PCR are useful in chronicity.

Gastrointestinal nematodes from buffalo in Minoufiya Governorate, Egypt with special reference to Bunostomum phlebotomum.

Gastrointestinal nematodes cause massive economic losses as an important impediment to the development of animals around the world. This study aimed to investigate the microscopical diagnosis of nematode parasites of buffalo from Minoufiya, Egypt and molecular characterization of Bunostomum phlebotomum. We examined 390 fecal samples with floatation and fecal culture techniques to recognize different genera of nematodes. The results revealed B. phlebotomum (2.56%), Strongyloides papillosus (3.85%), Toxocara vitulorum (7.69%), Haemonchus sp. (1.28%), and Dictyocaulus viviparus (1.28%). The recovered eggs and larvae of nematodes were identified as well as the adults of B. phlebotomum. Age-wise, sex-wise, and seasonal prevalences of the recovered nematodes were recorded. Sequence analysis of the ITS-2 gene of B. phlebotomum was highly identical (99-100%) to sequences from Australia and China and occurred in the same clade with B. trigoncephalum. In conclusion, the study presented the coprological survey of gastrointestinal nematodes, and the genetic characterization of B. phlebotomum from Minoufiya Governorate, Egypt for the first time.

Prevalence, hematological, serum biochemical, histopathology, and molecular characterization of Sarcoptes scabiei in naturally infected rabbits from Minoufiya Governorate, Egypt.

Sarcoptic mange is a highly contagious ectoparasitic disease that causes significant economic losses in the rabbit industry. The current study intended to reveal the infection rate, histopathology, and genetic characterization of Sarcoptes scabiei (S. scabiei) in naturally infected rabbits in Minoufiya governorate, Egypt. A total of 1120 rabbits were physically inspected for sarcoptic mange lesions and infections were confirmed microscopically. In addition, the various hematologic and serum biochemical parameters in naturally infected and non-infected rabbits were evaluated. A histopathological examination was performed. Genomic DNA was isolated from skin scraping samples and amplified using PCR primers targeting the ITS-2 region and Cox1 and Actin genes, which were then sequenced and phylogenetically analyzed. The overall prevalence of S. scabiei was 5.98%. Although the infection was higher in females than males, the analysis showed no statistically significant difference. White blood cells, lymphocytes, liver enzymes (GOT and GPT), urea, total antioxidant capacity, glutathione peroxidase, and malondialdehyde dramatically increased whereas RBCs, Hb, and MCV significantly decreased. There were epidermal thickening and hyperkeratosis, inflammation, and homogenous faint pink edematous lesions, and the S. scabiei was attached to the stratum corneum and/or burrowing through it, causing tunnels. PCR and sequence analysis of the ITS-2 region and Cox1 and Actin genes showed that the sequences in the present study were highly identical to the homologous sequences from several countries and confirmed that the mite was S. scabiei. This study presented the first molecular characterization of S. scabiei in rabbits from Minoufiya Governorate, Egypt.

Identification and Characterization of P0 Protein as a Vaccine Candidate Against Babesia divergens, Blood Parasite of Veterinary and Zoonotic Importance.

The molecular identification and antigenic characterization of P0 protein in Babesia divergens, a blood parasite of veterinary and zoonotic importance, were carried out in this study for use in developing subunit vaccines against B. divergens infection. Recombinant protein encoding P0 (BdP0) was developed in Escherichia coli, and its antiserum was generated in mice for further molecular characterization. Anti-rBdP0 serum had a specific interaction with the corresponding legitimate B. divergens protein, as confirmed by Western blotting and indirect fluorescent antibody tests. ELISA was used to assess the immunogenicity of BdP0 in a group of 68 bovine field samples, and significant immunological reactivity was found in 19 and 20 positive samples of rBdp0 and B. divergens lysate, respectively. The in vitro growth of B. divergens cultures treated with anti-rBdP0 serum was significantly inhibited (p < 0.05). Furthermore, after 6 h of incubation with 2 mg/ml anti-rBdP0 serum, the ability of pre-incubated free merozoites to invade bovine erythrocytes was reduced by 59.88%. The obtained data suggest the possible use of rBdP0 as diagnostic antigen and may serve as a vaccine candidate against babesiosis caused by B. divergens either in animal or human.

Infection rate and molecular characterization of Echidnophaga gallinacea in chickens from Matrouh Governorate, Egypt.

Echidnophaga gallinacea is the sticktight flea of chickens. It causes dermatitis and ulcers in the skin and carries some disease-causing agents such as Rickettsia and Bartonella. This study was conducted to detect the infection rate and elucidate the molecular characterization of E. gallinacea in chickens from El-Dabaa City, Matrouh Governorate, Egypt. The fleas were collected from infected chickens and identified morphologically. The internal transcribed spacer-1 (ITS-1) gene PCR method was used for molecular characterization. Based on the morphology, the collected fleas were confirmed as E. gallinacea. The overall infection rate was 5%, with 4.5% in female and 10% in male chickens. ITS-1 PCR revealed a specific band of 488 bp. The ITS-1 gene sequence from Egypt occurred in the same phylogenetic clade as that from Cameroon, with a percentage identity of 98.47%.

Strongylus vulgaris: Infection rate and molecular characterization from naturally infected donkeys at Sadat City, Egypt.

Strongylus vulgaris has high pathogenicity to equines. It causes aneurysm and thrombosis in the arteries particularly an anterior mesenteric artery, that is fatal to equines. In this study, we aimed to diagnose microscopically the natural infection of donkeys with Strongylus vulgaris from Sadat City, Minoufiya Governorate, Egypt. Fecal egg culture was used after the diagnosis of strongyle eggs to identify the species. Hematological and biochemical parameters were assessed. Adult worms were collected after post mortem examination of the infected animal. The sequence of ITS-2 was used to confirm the species of the parasite. The infection rate was 15.85% using the microscopical examination. The larval culture confirmed the infection with strongyle eggs as Strongylus vulgaris larvae. The sequence of ITS-2 was highly identical (about 95%) to sequences from Germany, China, and Turkey and occurred in the same genetic clade with the sequence from Germany. In conclusion, the study presented the diagnosis, the changes in the hematological and biochemical parameters in the infected animals, and the genetic characterization of Strongylus vulgaris from Sadat City, Minoufiya Governorate Egypt for the first time.

Infection rate and biochemical characterization of Cysticercus tenuicollis from sheep in Minoufiya governorate, Egypt.

Cysticercus tenuicollis, the larval stage of Taenia hydatigenia, infects sheep and causes economic losses due to condemnation of infected organs. This study was designed to report the infection rate, risk factors, biochemical, and molecular characterization of Cysticercus tenuicollis in sheep from Ashmoun, Minoufiya, Egypt. The infection rate was 18%. The age was a risk factor for infection where there was a significant difference in infection rate between sheep more than 3 years and sheep under 3 years of age. There was no significant difference between infection in male and female groups. The liver had the highest organ distribution followed by omentum. Biochemical analysis of the cyst fluid showed some variations in the levels of ALT, AST, triglycerides, cholesterol, total protein, urea nitrogen, calcium, sodium, chromium, potassium than the levels identified in Algeria, Iraq, and Iran. PCR and sequence analysis of cox1 and ssrRNA showed that the sequences from Minoufiya, Egypt were highly identical to the related ones from several countries and confirmed the cyst is Cysticercus tenuicollis. This study reported the infection rate, risk factors, biochemical analysis, and molecular characterization of Cysticercus tenuicollis in sheep from Minoufiya, Egypt.

An epidemiological survey of Theileria equi parasite in donkeys (Equus asinus) in Egypt.

In the present study, we conducted an epidemiological survey of Theileria equi, with sequencing analysis of the PCR product using blood-DNA samples collected from donkeys (n = 149) reared in different Egyptian provinces in Lower Egypt (Menoufia and Mersa Matruh) and middle Egypt (Giza). All animals were tested for the presence of T. equi parasite using species-specific PCR assay targeting the Equi merozoite antigen-1 (EMA-1). Nine- (6.04%) samples were positive for T. equi. The highest positive rate for infection was detected in Giza zoological garden (10.16%). Egyptian EMA-1 gene sequence exhibited a high identity with gene sequence from Italy, Japan, South Africa, Indian and Israel, the Palestinian Authority. In conclusion, data presented here revealed for the presence of T. equi in donkeys in two provinces of Egypt either in form of acute infection or carriers. These findings have economic significance and indicate the importance of introducing effective prevention and control strategies throughout Egypt to minimize the prevalence of equine piroplasmosis caused by T. equi.

Myrrh Oil in Vitro Inhibitory Growth on Bovine and Equine Piroplasm Parasites and Babesia microti of Mice.

The present experimental study was conducted for the assessment of the efficacy of in vitro inhibition of myrrh oil on the propagation of Babesia bovis, B. divergens, B. bigemina, Theileria equi, and B. caballi and in vivo efficacy on B. microti in mice through fluorescence assay based on SYBR green I. The culture of B. divergens B. bovis and was used to evaluate the in vitro possible interaction between myrrh oil and other commercial compound, such as pyronaridine tetraphosphate (PYR), diminazene aceturate (DA), or luteolin. Nested-polymerase chain reaction protocol using primers of the small-subunit rRNA of B. microti was employed to detect any remnants of DNA for studied parasitic species either in blood or tissues. Results elucidated that; Myrrh oil significantly inhibit the growth at 1% of parasitic blood level for all bovine and equine piroplasm under the study. Parasitic regrowth was inhibited subsequently by viability test at 2 µg/mL for B. bigemina and B. bovis, and there was a significant improvement in the in vitro growth inhibition by myrrh oil when combined with DA, PYR, and luteolin. At the same time; mice treated with a combination of myrrh oil/DA showed a higher inhibition in emitted fluorescence signals than the group that challenged with 25 mg/kg of diminazene aceturate at 10 and 12 days post-infection. In conclusion, this study has recommended the myrrh oil to treat animal piroplasmosis, especially in combination with low doses of DA.

Inhibitory effects of fluoroquinolone antibiotics on Babesia divergens and Babesia microti, blood parasites of veterinary and zoonotic importance.

AIM: This study aimed to evaluate the inhibitory effects of fluoroquinolone antibiotics, including enrofloxacin, enoxacin, trovafloxacin, norfloxacin, and ofloxacin, on the in vitro and in vivo growth of Babesia divergens and Babesia microti parasites, respectively. MATERIALS AND METHODS: The in vitro and in vivo inhibitory effects of fluoroquinolone antibiotics against B. divergens and B. microti, respectively were evaluated using fluorescence-based assay. Additionally, combination therapies of highly effective fluoroquinolone antibiotics (enrofloxacin, enoxacin, and trovafloxacin) with diminazene aceturate, luteolin, or pyronaridine tetraphosphate were tested on the in vitro cultures of B. divergens. RESULTS: Enrofloxacin, trovafloxacin, and enoxacin were the most effective fluoroquinolones against the in vitro growth of B. divergens, followed by norfloxacin and ofloxacin. Furthermore, a combination of enoxacin or trovafloxacin with either diminazene aceturate, luteolin, or pyronaridine tetraphosphate significantly enhanced the inhibitory effect on the growth of B. divergens in in vitro cultures. In mice infected by B. microti, enoxacin and diminazene aceturate combination therapy exhibited a potential antibabesial effect. CONCLUSION: These results suggest that safe and cheap fluoroquinolone, such as enoxacin, might be used for the treatment of clinical cases caused by Babesia spp. in animals or humans.