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Abstract Background Theileria haneyi is one of the three known causative agents of equine piroplasmosis. While imidocarb is generally effective in the clearance of the highly pathogenic Theileria equi, it is ineffective in the treatment of T. haneyi. Moreover, co-infection with T. haneyi has been shown to impede the successful treatment of T. equi. Furthermore, tulathromycin and diclazuril have demonstrated inefficacy in eradicating T. haneyi. The absence of an effective therapeutic agent against this parasite represents a significant obstacle in managing equine piroplasmosis. Methods To address this issue, we evaluated the efficacy of buparvaquone in the treatment of T. haneyi in chronically infected horses. Results Our findings showed that treatment of horses with the recommended dose of 2.5 mg/kg of buparvaquone led to a rapid abatement of T. haneyi levels, to a level where the parasites were not detectable by nested PCR. Following treatment, the horses remained PCR negative for a minimum of seven weeks until recrudescence occurred. Subsequent re-administration of buparvaquone at an increased dosage of 6 mg/kg upon recrudescence failed to exert a theilericidal effect on T. haneyi. Throughout the treatment regimen, the hematological parameters of the horses and most components of the chemistry panel remained within the normal range, except for blood urea nitrogen levels, which fell below the normal range in certain instances. conclusions BPQ at 2.5 mg/kg and 6 mg/kg had a robust theilericidal effect but was ineffective in the clearance of the T. haneyi infection in persistently infected animals. Graphical Abstract

A 2-year-old female Assam Hill goat was presented with a clinical history of anorexia, fever, mild anemia, rough body coat, dehydration, tachycardia, dyspnea and swelling of palpable lymph nodes. Hematology revealed low hemoglobin, packed cell volume, red blood cell and thrombocyte count. Biochemical analysis showed increased serum concentration of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine and urea in comparison to the normal reference range. Microscopic examination showed intra-erythrocytic forms of Theileria species. Molecular and phylogenetic analysis of partial 18S rRNA gene sequence confirmed Theileria luwenshuni infection. The goat was treated with buparvaquone and oxytetracycline and recovered uneventfully. A three-month follow-up showed no recurrence. This study reveals the presence of T. luwenshuni in Assam, India and it should be considered in differential diagnosis and as one of the important pathogens of clinically sick goats. The present case report provides a rational approach to diagnosis and treatment for a goat infected with pathogenic T. luwenshuni in Assam, India. To our knowledge, the present communication describes about the first successful therapeutic management of pathogenic T. luwenshuni infection in a goat supported with molecular evidence from Assam, a north-eastern state of India. [ABSTRACT FROM AUTHOR]

Several miRNA-based studies on Theileria-transformed bovine cells have been conducted; however, the mechanism by which transformed cells exhibit uncontrolled proliferation is not yet fully understood. Therefore, it is necessary to screen more microRNAs that may play a role in the transformation process of host cells infected with Theileria annulata to better understand the transformation mechanisms of Theileria-infected cells. RNA sequencing was used to analyze miRNAs expression in the host bovine lymphocytes infected with T. annulata at different time points after buparvaquone (BW720) treatment and DMSO treatment (control groups). Differential miRNAs related to cell proliferation and apoptosis were identified through comparison with gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and a regulatory network of miRNA-mRNA was constructed. In total, 272 differentially expressed miRNAs were found at 36, 60 and 72 h. The miRNAs change of bta-miR-2285t, novel-miR-622, bta-miR-2478, and novel-miR-584 were significant. Analysis of 27 of these co-differential expressed miRNAs revealed that 15 miRNAs were down-regulated and 12 miRNAs were up-regulated. A further analysis of the changes in the expression of each of these 27 miRNAs in the three datasets suggested that bta-miR-2285t, bta-miR-345-5p, bta-miR-34a, bta-miR-150, and the novel-miR-1372 had significantly changed. Predicted target genes for these 27 miRNAs were analyzed by KEGG and the results demonstrated that EZR, RASSF, SOCS1 were mainly enriched in the signaling pathway microRNAs in cancer. MAPKAPK2, RELB, FLT3LG, and GADD45B were mainly enriched in the MAPK signaling pathway, and some genes were enriched in Axon guidance. This study has provided valuable information to further the understanding of the regulatory function of miRNAs in the host microenvironment and host-parasite interaction mechanisms. [ABSTRACT FROM AUTHOR]

Background: Theileria haneyi is one of the three known causative agents of equine piroplasmosis. While imidocarb is generally effective in the clearance of the highly pathogenic Theileria equi, it is ineffective in the treatment of T. haneyi. Moreover, co-infection with T. haneyi has been shown to impede the successful treatment of T. equi. Furthermore, tulathromycin and diclazuril have demonstrated inefficacy in eradicating T. haneyi. The absence of an effective therapeutic agent against this parasite represents a significant obstacle in managing equine piroplasmosis. Methods: To address this issue, we evaluated the efficacy of buparvaquone in the treatment of T. haneyi in chronically infected horses. Results: Our findings showed that treatment of horses with the recommended dose of 2.5 mg/kg of buparvaquone led to a rapid abatement of T. haneyi levels, to a level where the parasites were not detectable by nested PCR. Following treatment, the horses remained PCR negative for a minimum of seven weeks until recrudescence occurred. Subsequent re-administration of buparvaquone at an increased dosage of 6 mg/kg upon recrudescence failed to exert a theilericidal effect on T. haneyi. Throughout the treatment regimen, the hematological parameters of the horses and most components of the chemistry panel remained within the normal range, except for blood urea nitrogen levels, which fell below the normal range in certain instances. conclusions: BPQ at 2.5 mg/kg and 6 mg/kg had a robust theilericidal effect but was ineffective in the clearance of the T. haneyi infection in persistently infected animals. [ABSTRACT FROM AUTHOR]

Introduction: Theileria orientalis, an economically significant tick-borne hemoparasite, infects cattle globally. The T. orientalis Ikeda genotype, transmitted by Haemaphysalis longicornis ticks, is associated with clinical manifestations characterized by anemia, abortions, and mortality, although subclinical infections prevail. Despite the common occurrence of subclinical infections, therapeutic interventions targeting T. orientalis Ikeda in such cases are currently lacking, impeding effective parasite control measures. To address this critical knowledge gap, we assessed the efficacy of buparvaquone (BPQ) in eliminating the T. orientalis Ikeda, US isolate, in sub-clinically infected cattle. Methods: Twelve sub-clinically infected calves, identified by the presence of T. orientalis in peripheral blood alongside the absence of fever and anemia, were enrolled in the study. Six calves received two treatments of the BPQ label dose (2.5mg/kg) at a 48-h interval, while additional three calves received the drug at a dosage of 6mg/kg following the same regimen. Three untreated calves served as controls. Results and discussion: Endpoint and quantitative PCR analyses revealed that BPQ exerted a transient effect on T. orientalis parasitemia. Parasites remained undetectable in peripheral blood until weeks 4 and 11 post-treatment in animals administered 2.5 mg/kg and 6 mg/kg of BPQ, respectively. Intriguingly, following recrudescence, administering 6 mg/kg to animals previously treated with 2.5 mg/kg did not result in a reduction in parasite load. Pharmacokinetic analysis data suggested that escalating the dosage led to a less than proportional increase in serum concentrations of BPQ. Moreover, a significant yet reversible decrease (p < 0.05) in blood urea nitrogen was observed in animals treated with the drug, irrespective of the dosage. Despite parasitemia relapse, animals treated with 6 mg/kg BPQ exhibited a noteworthy decrease (p < 0.05) in IgG levels specific to the T. orientalis major piroplasm surface protein compared to controls and animals treated with 2.5 mg/kg of the drug. Conclusion: BPQ did not demonstrate efficacy in clearing subclinical T. orientalis Ikeda infection. Future investigations are warranted to explore innovative therapeutic modalities that, in synergy with vaccines and diagnostic assays, can facilitate the development of comprehensive programs aimed at controlling and eradicating this parasite. [ABSTRACT FROM AUTHOR]

A 2-year-old female Assam Hill goat was presented with a clinical history of anorexia, fever, mild anemia, rough body coat, dehydration, tachycardia, dyspnea and swelling of palpable lymph nodes. Hematology revealed low hemoglobin, packed cell volume, red blood cell and thrombocyte count. Biochemical analysis showed increased serum concentration of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine and urea in comparison to the normal reference range. Microscopic examination showed intra-erythrocytic forms of Theileria species. Molecular and phylogenetic analysis of partial 18S rRNA gene sequence confirmed Theileria luwenshuni infection. The goat was treated with buparvaquone and oxytetracycline and recovered uneventfully. A three-month follow-up showed no recurrence. This study reveals the presence of T. luwenshuni in Assam, India and it should be considered in differential diagnosis and as one of the important pathogens of clinically sick goats. The present case report provides a rational approach to diagnosis and treatment for a goat infected with pathogenic T. luwenshuni in Assam, India. To our knowledge, the present communication describes about the first successful therapeutic management of pathogenic T. luwenshuni infection in a goat supported with molecular evidence from Assam, a north-eastern state of India.

Introduction: B. bovis is an apicomplexan parasite responsible for bovine babesiosis, a tick-borne disease with a worldwide impact. The disease remains inefficiently controlled, and few effective drugs, including imidocarb dipropionate (ID), are currently available in endemic areas. The objective of this study was to evaluate whether buparvaquone (BPQ), a drug currently used to treat cattle infected with the Babesia-related Theileria spp. parasites, could be active against Babesia parasites. Herein, we compared the effect of ID and BPQ on B. bovis growth in vitro erythrocyte culture. Methods: We compared the effect of ID and BPQ on the culture-adapted Texas T2Bo strain of B. bovis. In vitro cultured parasites were incubated with ID and BPQ at two starting parasitemia levels (PPE), 0.2% and 1%. In vitro cultured parasites were treated with ID or BPQ at concentrations ranging from 10 to 300 nM, during 4 consecutive days. Parasitemia levels were daily evaluated using microscopic examination. Data was compared using the independent Student's t-test. Results and Discussion: Both ID and BPQ significantly inhibited (p < 0.05) the growth of B. bovis, regardless of the initial parasitemia used. At 1% parasitemia, BPQ had lower calculated inhibitory concentration 50 (IC50: 50.01) values than ID (IC50: 117.3). No parasites were found in wells with 0.2% starting parasitemia, treated previously with 50 nM of BPQ or ID, after 2 days of culture without drugs. At 1% parasitemia, no parasite survival was detected at 150 nM of BPQ or 300 nM of ID, suggesting that both drugs acted as babesiacidals. Conclusion: Overall, the data suggests that BPQ is effective against B. bovis and shows a residual effect that seems superior to ID, which is currently the first-line drug for treating bovine babesiosis globally. [ABSTRACT FROM AUTHOR]

Ocular toxoplasmosis is likely the most common cause of infectious posterior uveitis worldwide. CXCL10 chemokine has an important role in the maintenance of the T-cell response and the control of Toxoplasma gondii in the eye during chronic infection. Drugs that can modulate the chemokine activity could be effective against the parasite. In this work, CXCL10 local retinal expression was investigated in a diabetic mouse model with ocular toxoplasmosis for the first time. In addition, the efficacy of naphthoquinones and quinolones was compared to spiramycin (SP) in treating the infection and modulating the chemokine expression. Our results revealed that chloroquine (CQ) achieved the best results regarding the reduction of cerebral cyst burden (84.36%), improving the retinal histopathological changes, cellular infiltrates, and vasculitis significantly (P < 0.005), and balancing the strong CXCL10 expression caused by the infection. Buparvaquone-treated mice showed a significant percentage of reduction of brain cysts (76.25%), moderate improvement of histopathology, and mild to moderate CXCL10 expression. While SP showed the least efficacy against the parasite in the eye in the form of mild improvement of histopathological changes and downregulation of retinal chemokine expression with the least reduction rate of cerebral parasitic burden (57%). In conclusion, Optimal control of pathogens probably needs a balanced immune response with an optimum expression of chemokines. So, targeting the modulation of retinal CXCL10 may eventually be beneficial in the management of ocular toxoplasmosis plus its potential to act as a marker for predictive local immunological response during the infection. [ABSTRACT FROM AUTHOR]

IntroductionTheileria orientalis, an economically significant tick-borne hemoparasite, infects cattle globally. The T. orientalis Ikeda genotype, transmitted by Haemaphysalis longicornis ticks, is associated with clinical manifestations characterized by anemia, abortions, and mortality, although subclinical infections prevail. Despite the common occurrence of subclinical infections, therapeutic interventions targeting T. orientalis Ikeda in such cases are currently lacking, impeding effective parasite control measures. To address this critical knowledge gap, we assessed the efficacy of buparvaquone (BPQ) in eliminating the T. orientalis Ikeda, US isolate, in sub-clinically infected cattle.MethodsTwelve sub-clinically infected calves, identified by the presence of T. orientalis in peripheral blood alongside the absence of fever and anemia, were enrolled in the study. Six calves received two treatments of the BPQ label dose (2.5 mg/kg) at a 48-h interval, while additional three calves received the drug at a dosage of 6 mg/kg following the same regimen. Three untreated calves served as controls.Results and discussionEndpoint and quantitative PCR analyses revealed that BPQ exerted a transient effect on T. orientalis parasitemia. Parasites remained undetectable in peripheral blood until weeks 4 and 11 post-treatment in animals administered 2.5 mg/kg and 6 mg/kg of BPQ, respectively. Intriguingly, following recrudescence, administering 6 mg/kg to animals previously treated with 2.5 mg/kg did not result in a reduction in parasite load. Pharmacokinetic analysis data suggested that escalating the dosage led to a less than proportional increase in serum concentrations of BPQ. Moreover, a significant yet reversible decrease (p

Introduction:B. bovis is an apicomplexan parasite responsible for bovine babesiosis, a tick-borne disease with a worldwide impact. The disease remains inefficiently controlled, and few effective drugs, including imidocarb dipropionate (ID), are currently available in endemic areas. The objective of this study was to evaluate whether buparvaquone (BPQ), a drug currently used to treat cattle infected with the Babesia-related Theileria spp. parasites, could be active against Babesia parasites. Herein, we compared the effect of ID and BPQ on B. bovis growth in vitro erythrocyte culture.Methods:We compared the effect of ID and BPQ on the culture-adapted Texas T2Bo strain of B. bovis. In vitro cultured parasites were incubated with ID and BPQ at two starting parasitemia levels (PPE), 0.2% and 1%. In vitro cultured parasites were treated with ID or BPQ at concentrations ranging from 10 to 300 nM, during 4 consecutive days. Parasitemia levels were daily evaluated using microscopic examination. Data was compared using the independent Student’s t-test.Results and discussion:Both ID and BPQ significantly inhibited (p < 0.05) the growth of B. bovis, regardless of the initial parasitemia used. At 1% parasitemia, BPQ had lower calculated inhibitory concentration 50 (IC50: 50.01) values than ID (IC50: 117.3). No parasites were found in wells with 0.2% starting parasitemia, treated previously with 50 nM of BPQ or ID, after 2 days of culture without drugs. At 1% parasitemia, no parasite survival was detected at 150 nM of BPQ or 300 nM of ID, suggesting that both drugs acted as babesiacidals.Conclusion:Overall, the data suggests that BPQ is effective against B. bovis and shows a residual effect that seems superior to ID, which is currently the first-line drug for treating bovine babesiosis globally.

Theileria annulata is an intracellular haemoprotozoan transmitted by ticks and its prevalence is reported mostly in the tropical region of the world. Tropical theileriosis mainly treated with Buparvaquone and tetracycline along with supportive therapy. Since there are reports on the development of resistance against buparvaquone so for that present study was carried out to evaluate the antitheilerial drug regime against the local strain of theileriosis. The present study on antitheilerial drug response was done to compare the combination therapy (Buparvaquone with tetracycline) and Buparvaquone only. A total of 18 Buffalo were selected from Ayodhya a Eastern Plane Zone (EPZ) of Uttar Pradesh and separated into three groups each with 6 animals. Treatment Group I and Group II were comprised of infected animals and group III included the healthy animals only. Before the experiment, at zero-day all 18 animals were screened for Theileria annulata infection by microscopic examination, and confirmed by Tams1 gene-based Polymerase Chain Reaction (PCR). Further, the same procedure followed for the blood for the samples collected on different days of post-treatment. Group I animal received the dose of buparvaquone @2.5mg/kg.b.wt. i.m., whereas the group II animals received the dose of buparvaquone @2.5mg/kg.b.wt. i.m., with oxytetracycline @20mg/kg.b.wt. i.v. twice within 48 hours. Hematological (Hb, PCV, TEC, DLC) and biochemical variables (total serum protein, albumin, blood urea nitrogen, AST, ALT) were assessed before and after treatment. After treatment the level of haemato-biochemical parameters restored early in the group II treated with combination therapy of Buparvaquone with tetracycline than in group I treated with Buparvaquone only. Moreover, the results of the treatment and control groups were significantly (<0.05) different with each other. The present study revealed that the treatment of Theileria annulata infected Buffalos with the combination of buparvaquone and oxytetracycline provides slightly better recovery than the treatment with buparvaquone only (group I). However, treatment with Buparvaquone or combination with tetracycline repeated after 48 hours both worked well against tropical theileriosis prevalence. [ABSTRACT FROM AUTHOR]

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Control of tropical theileriosis, caused by the apicomplexan Theileria annulata, depends on the use of a single drug, buparvaquone, the efficacy of which is compromised by the emergence of resistance. The present study was undertaken to improve understanding of the role of mutations conferring buparvaquone resistance in T. annulata, and the effects of selection pressures on their emergence and spread. First, we investigated genetic characteristics of the cytochrome b locus associated with buparvaquone resistance in 10 susceptible and 7 resistant T. annulata isolates. The 129G (GGC) mutation was found in the Q01 binding pocket and 253S (TCT) and 262S (TCA) mutations were identified within the Q02 binding pocket. Next, we examined field isolates and identified cytochrome b mutations 129G (GGC), 253S (TCT) and 262S (TCA) in 21/75 buffalo-derived and 19/119 cattle-derived T. annulata isolates, providing evidence of positive selection pressure. Both hard and soft selective sweeps were identified, with striking differences between isolates. For example, 19 buffalo-derived and 7 cattle-derived isolates contained 129G (GGC) and 253S (TCT) resistance haplotypes at a high frequency, implying the emergence of resistance by a single mutation. Two buffalo-derived and 12 cattle-derived isolates contained equally high frequencies of 129G (GGC), 253S (TCT), 129G (GGC)/253S (TCT) and 262S (TCA) resistance haplotypes, implying the emergence of resistance by pre-existing or recurrent mutations. Phylogenetic analysis further revealed that 9 and 21 unique haplotypes in buffalo and cattle-derived isolates were present in a single lineage, suggesting a single origin. We propose that animal migration between farms is an important factor in the spread of buparvaquone resistance in endemic regions of Pakistan. The overall outcomes will be useful in understanding how drug resistance emerges and spreads, and this information will help design strategies to optimise the use and lifespan of the single most drug use to control tropical theileriosis.

Cryptosporidium parvum is a zoonotic protozoan that causes neonatal calf diarrhea common in the world. Buparvaquone, which is known to have very positive effects on oocysts, which are the main reserve in the spread of infection, has not yet been investigated for its hemato-biochemical aspect in calves with cryptosporidiosis. In the present study, the effect of buparvaquone on renal and hepatic functions in naturally infected newborn calves with cryptosporidiosis was investigated. A group was formed for this study using only a total of ten calves (n=10) naturally infected with cryptosporidiosis. Buparvaquone 2.5 mg/kg intramuscular injection was administered as a single dose to all calves in the group. Laboratory analyses and statistical calculations of blood and serum samples taken on the specified days were made. According to the results we obtained in the study, it was observed that the renal and hepatic effects of the drug after a single dose injection of buparvaquone to calves with neonatal cryptosporidiosis remained within normal limits, similar to the methods safely applied in the treatment of cryptosporidiosis in the field. [ABSTRACT FROM AUTHOR]

Bovine tropical theileriosis is one of the potentially fatal disease of dairy cattle, which is caused by hemoparasite Theleria annulata. About seven years old cross-bred cow was presented with complaint of pyrexia, inappetance, lacrimation and ocular swelling since last 5 days. The clinical examination showed elevated rectal temperature (39.4 0C), mild enlarged pre-scapular lymph nodes, bilateral bulging of temporal fossa, protruded pale and icteric conjunctivae of the eyes with lacrimation and presence of ticks on body. The case was suspected for haemoprotozoan disease. Blood and serum sample were collected for hematological, blood smear examination and molecular examination (PCR), and biochemical analysis respectively. Microscopic examination of blood smear revealed intra-erythrocytic signet ring shaped periplasm of Theileria annulata. Hemato-biochemical examination revealed anemia, hypoproteinemia, hypoalbuminemia and jaundice. Further, PCR assay was done using T. annulata-specific primer pair, Cyto b1 gene targeting the amplicon of 312 bp showed specific band on Gel-electrophoresis. Therapeutic regimen was started with Buparvaquone @ 2.5 mg/kg body weight IM single dose followed by Oxytetracycline @ 10 mg/kg body weight IV in 500 ml of NS for 5 days and Prednisolone @ 0.25 mg/kg body weight IM for 3 days along with supportive therapy. The cattle well responded to the therapy and complete regression of ocular signs was observed within one week of treatment. [ABSTRACT FROM AUTHOR]

Bovine tropical theileriosis is one of the potentially fatal disease of dairy cattle, which is caused by hemoparasite Theleria annulata. About seven years old cross-bred cow was presented with complaint of pyrexia, inappetance, lacrimation and ocular swelling since last 5 days. The clinical examination showed elevated rectal temperature (39.4 0C), mild enlarged pre-scapular lymph nodes, bilateral bulging of temporal fossa, protruded pale and icteric conjunctivae of the eyes with lacrimation and presence of ticks on body. The case was suspected for haemoprotozoan disease. Blood and serum sample were collected for hematological, blood smear examination and molecular examination (PCR), and biochemical analysis respectively. Microscopic examination of blood smear revealed intra-erythrocytic signet ring shaped periplasm of Theileria annulata. Hemato-biochemical examination revealed anemia, hypoproteinemia, hypoalbuminemia and jaundice. Further, PCR assay was done using T. annulata-specific primer pair, Cyto b1 gene targeting the amplicon of 312 bp showed specific band on Gel-electrophoresis. Therapeutic regimen was started with Buparvaquone @ 2.5 mg/kg body weight IM single dose followed by Oxytetracycline @ 10 mg/kg body weight IV in 500 ml of NS for 5 days and Prednisolone @ 0.25 mg/kg body weight IM for 3 days along with supportive therapy. The cattle well responded to the therapy and complete regression of ocular signs was observed within one week of treatment.

Buparvaquone 1a is a predominant anti-protozoal drug, it belongs to the pharmacologically active class of hydroxynaphthoquinones. There are numerous publications on the wide therapeutic applications of 1a, but only a few approaches were reported towards its synthesis. Most of the prior arts report the synthesis from expensive raw materials with low yield, whereas only a few involves the use of readily available and less expensive raw materials with moderate to better yield. The present review work covers the developments on therapeutic applications of 1a along with the synthetic approaches disclosed till date. [ABSTRACT FROM AUTHOR]

Theileria annulata (T. annulata) is intra-erythrocytic protozoan parasite which is more prevalent in tropical and sub-tropical countries. It has a significant economic impact on the productivity of the dairy industry, and buparvaquone is used to treat infected animals in the prevalent regions of the world. Systematically, buparvaquone targets the cyto-b gene to break the electron transport chain (ETC) and Theileria annulata peptidyl-prolyl isomerase 1 (TaPIN1) gene to destabilize transcription factor JUN (c-JUN) to inhibit proliferation of infected cells, which ultimately leads to the death of T. annulata. The reported studies on drug resistance is due to inappropriate drug application, evolutionary characteristics of the cytochrome b (cyto-b) gene and oncogenic signaling pathways gene (TaPIN1) make the parasite resistant against buparvaquone. Hence, this systematic review was designed to find out non-synonymous mutation in genes (cyto-b and TaPIN1) responsible for drug resistance reported from Tunisia, Turkey, Egypt, Sudan, Iran, Pakistan, China and Germany with reference to the T. annulata Ankara strain of cyto-b (accession no. XM_949625.1) and TaPIN1 (accession no. TA18945) wild type genes. Non-synonymous point mutations were found in cyto-b (Q 01 at 130–148 and Q 02 at 253–262 regions) and TaPIN1 (A53P and A53T) genes. These point mutations are responsible for developing buparvaquone resistance against T. annulata infection. These genes can be used as biomarkers for the identification of drug resistance in any endemic area. To avoid the complication of drug resistance, development of genetically resistant cattle breeds, potent vaccines and anti-theilerial drugs (Trifloxystrobin and anti-cancerous) are currently required to control proliferating economically important T. annulata parasites. • Buparvaquone is the drug of choice for tropical theileriosis. • This has developed resistance against buparvaquone. • Two biomarkers are being used for molecular identification of drug resistance in tropical theileriosis. [ABSTRACT FROM AUTHOR]

Control of tropical theileriosis, caused by the apicomplexan Theileria annulata , depends on the use of a single drug, buparvaquone, the efficacy of which is compromised by the emergence of resistance. The present study was undertaken to improve understanding of the role of mutations conferring buparvaquone resistance in T. annulata , and the effects of selection pressures on their emergence and spread. First, we investigated genetic characteristics of the cytochrome b locus associated with buparvaquone resistance in 10 susceptible and 7 resistant T. annulata isolates. The 129G (GGC) mutation was found in the Q 01 binding pocket and 253S (TCT) and 262S (TCA) mutations were identified within the Q 02 binding pocket. Next, we examined field isolates and identified cytochrome b mutations 129G (GGC), 253S (TCT) and 262S (TCA) in 21/75 buffalo-derived and 19/119 cattle-derived T. annulata isolates, providing evidence of positive selection pressure. Both hard and soft selective sweeps were identified, with striking differences between isolates. For example, 19 buffalo-derived and 7 cattle-derived isolates contained 129G (GGC) and 253S (TCT) resistance haplotypes at a high frequency, implying the emergence of resistance by a single mutation. Two buffalo-derived and 12 cattle-derived isolates contained equally high frequencies of 129G (GGC), 253S (TCT), 129G (GGC)/253S (TCT) and 262S (TCA) resistance haplotypes, implying the emergence of resistance by pre-existing or recurrent mutations. Phylogenetic analysis further revealed that 9 and 21 unique haplotypes in buffalo and cattle-derived isolates were present in a single lineage, suggesting a single origin. We propose that animal migration between farms is an important factor in the spread of buparvaquone resistance in endemic regions of Pakistan. The overall outcomes will be useful in understanding how drug resistance emerges and spreads, and this information will help design strategies to optimise the use and lifespan of the single most drug use to control tropical theileriosis. [Display omitted] • First link between genetic mutations and buparvaquone drug resistance in T. annulata. • The data will help to design strategies to optimise the use and lifespan of the single most drug use to control tropical theileriosis. [ABSTRACT FROM AUTHOR]

Theileria annulata schizont-infected host cells in culture in vitro show unlimited proliferation similar to tumor cells; thus far, T. annulata and T. parva are the only eukaryotes that have been found to transform mammalian cells (immortalized). The transformation of these cells is reversible; when the parasite is eliminated in transformed cells by buparvaquone (BW720c), the host cells show normal growth and apoptosis. TFG is a tropomyosin-receptor kinase fused gene that is conserved among many species and is an important proto-oncogene. In this study, the bovine TFG gene was amplified by PCR from the cDNA of T. annulata schizont-transformed cells, cloned into the pGEX-4T-1 vector and expressed in Escherichia coli BL21 (DE3). After purification, the fusion protein was injected into rabbits to produce polyclonal antibodies. Using T. annulata-transformed cells together with BW720c treatment to kill the parasite, we aimed to identify changes in TFG gene expression by real-time PCR and Western blotting. The results showed that the bovine TFG gene was ~582 bp in size; SDS-PAGE analysis showed that the fusion protein was expressed in BL21 (DE3) cells with a molecular mass of 48 kD, and Western blotting indicated that the polyclonal antibodies could react with bovine TFG proteins from T. annulata-transformed cells and showed high specificity. Compared with that in the control group, the transcription level of the host TFG gene decreased significantly in the BW720c test group, and the expression of host tumor-related TFG protein decreased sharply after 72 h of drug treatment, suggesting that the TFG protein expression in transformed cells was directly related to T. annulata. This finding laid a foundation for further study on the interaction between T. annulata and host cells.

This study aimed to investigate the evolutionary status of T. annulata in Al-Diwaniyah province, Iraq. In this study, the clinical examination of 50 infected animals was performed with blood sample collection (2.5ml per animal), and drug targets cytochrome b, a vital component of the electron transfer chain in the mitochondria of the protozoan, cytb gene was targeted using a polymerase chain reaction (PCR) procedure. Also, 18S rRNA gene as a molecular target for the PCR and a partial gene sequencing (PGS) were included. The PCR that involved using the 18S rRNA and cytb genes as genetic targets revealed amplification of the targeted pieces at 620bp and 1092bp, respectively, in all tested samples. The18S rRNA gene sequence of local T. annulata isolates were aligned with global reference strains for T. annulata recorded in the GenBank. The local strains were close, 100%, in their identity to isolates from Iran, Turkey, and Pakistan; however, they were 99% similar to a nucleotide sequences from India and Bangladesh. Diseased calves showed clinical signs such as high fever (40.3-41.5°C), decreased appetite or in appetence, asymmetrical enlargement of superficial lymph nodes particularly the pre-scapular ones, some cases with diarrhea, pale or icteric mucus membrane of eyes, bulging eyes, lacrimation, ecchymotic hemorrhages on the sclera, incoordination, nervous signs (Dullness, depression, lethargy), salivation, and bloated young calves. The data observed from the present inspecting work may reveal genetic evolution in the local strains with others recorded in the GeneBank. This means that our local strains might have close relationships with some global strains.

Aim: The aim of this study was to determine the possible side effects in sheep of combined administration of imidocarb, buparvaquone, and oxytetracycline, which are antiprotozoal drugs against bloodborne parasites, at the maximum dose and treatment period. . Materials and Methods: Imidocarb (2.4 mg/kg), buparvaquone (2.5 mg/kg), and oxytetracycline (20 mg/kg) were administered simultaneously by intramuscular injection to 10 sheep, and a second combined dose was administered 3 days later. Blood samples were taken before (0 control) and at 0.5, 1, 2, 3, 4, 5, and 6 days after drug administrations. Malondialdehyde, 8-hydroxy-2?-deoxyguanosine, troponin I, and creatine kinase-MB isoenzyme levels were determined with ELISA reader, and cardiac, hepatic, and renal damage markers were measured with autoanalyzer. Blood gas and hemogram parameters were also determined. Results: No oxidative stress (p>0.05) was observed in the sheep, whereas increased (p Conclusion: The combination of imidocarb, buparvaquone, and oxytetracycline have not cause serious oxidative damage, cardiotoxicity, and nephrotoxicity in sheep, but this combination may show transient effects on liver function and blood gas and hemogram values. Therefore, caution should be exercised when using this combination in infected sheep.

Theileriosis is an economically important haemoprotozoan disease of cattle caused by Theileria spp. It causes production losses and mortality in dairy industry. A total of 135 cattle suspected of theileriosis were screened based on clinical signs and blood smear examination (Giemsa's stain). Out of them, 64 animals were diagnosed with theileriosis. Twenty-four cattle out of them were selected and divided into four equal groups, viz., B, C, D, and E, each comprised of six animals for four different treatment protocols to evaluate their therapeutic efficacy, while group A of six healthy cattle served as control. The most common clinical findings of theileriosis infected cattle (n = 24) in descending order were anorexia (100%), dullness and depression, decreased milk production, weakness, salivation, enlarged pre-scapular lymph nodes, pale conjunctival mucous membrane, presence of ticks over the body, emaciation, nasal discharge, cessation of rumination, lacrimation, diarrhea and sternal recumbency (8.33%). The mean rectal temperature, heart rate, and respiratory rate were 104.5 ± 0.33° F, 79.5 ± 2.85/min and 42.83 ± 2.67/min, respectively. The haematological findings revealed decreased values of haemoglobin, packed cell volume, total erythrocytes count (TEC), lymphocytes, and mean corpuscular hemoglobin concentration (MCHC), and increased values of neutrophils, eosinophils, Mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) in theileriosis infected cattle as compared to healthy cattle. Biochemical findings revealed decreased values of total protein and albumin while increased total bilirubin, Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), and Gamma-glutamyl transferase (GGT) in theileriosis-infected cattle. These altered haemato-biochemical profiles were found to be normalized by the 14th day in different treatment groups. The therapeutic efficacy of Buparvaquone 2 dose (Group B) and Buparvaquone plus Oxytetracycline LA (Group D) was better (100.00%) than the Arteether (Group C, 66.66%) and Arteether plus Oxytetracycline LA (Group E, 83.33%) in theileriosis infected cattle, hence may be advocated to the field veterinarians for effective treatment of theileriosis in cattle. [ABSTRACT FROM AUTHOR]

A novel, simple, accurate, precise, economical and reliable ultraviolet spectrophotometric method has been developed for the estimation of buparvaquone in bulk and in pharmaceutical dosage form. The drug shows maximum absorption at 251 nm by using acetonitrile as solvent. The method was validated as stated in International Council for Harmonisation Q2 (R1) guidelines. It obeys Beer's law in the concentration range of 2-20 µg/ml with correlation coefficient of 0.998. The drug shows great accuracy close to 100 %. The method was found to be robust and precise as the relative standard deviation are less than 2 %. Limit of detection and limit of quantitation were found to be 0.60 µg/ml and 1.83 µg/ml respectively. From the results of specificity, the drug was found to be more degraded under alkaline, oxidative and photolytic conditions. The proposed method can be employed for the reliable quantification of buparvaquone in bulk and routine analysis of pharmaceutical formulations. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a tick-borne disease that caused by Theileria annulata , and leads to substantial economic impact in endemic area. Distinguishes to other piroplasms, Theileria is the only eukaryotic parasite could transform mammalian leukocytes. At present, buparvaquone is the most effective drug used for treatment of Theileria infection. However, frequently reported of failure treatment with buparvaquone for some T. annulata isolates. Mutation of TaPIN1 was reported to be the direct reason for failure of buparvaquone treatment. Through in vitro culture, a T. annulata isolate with a TaPIN1 mutation that is similar to the reported strain was recently identified in China. In order to understand the distribution of Theileria with mutation of TaPIN1 in China, here we developed a TaqMan probe-based real-time PCR technology to detect the mutated TaPIN1 gene. The specificity, sensitivity and reproducibility of the established TaqMan Real-time PCR method were evaluated, and field cattle blood samples collected from Xinjiang Uyghur Autonomous Region were used to test its application. Among 1683 samples, 335 samples were confirmed positive for T. annulata by traditional PCR method and 34 samples were positive for buparvaquone-resistant. The TaPIN1 gene of those 34 samples was sequenced and analyzed with the published gene sequences from NCBI database. The results showed that the sequence obtained from the present study has good consistency with those published sequences. In conclusion, the TaqMan probe-based real-time PCR targeting T. annulata mutated TaPIN1 gene was successfully established and can be used to detect clinical samples to investigation of buparvaquone-resistant parasites in Xinjiang region quickly and accurately, which will be useful for guiding clinical medicine application. • T. annulata resistant to buparvaquone was found in China where buparvaquone was forbidden to be used to treat cattle. • The mutation of TaPIN1 where most likely lead to buparvaquone-resist for Theileria annulata was detected. • Established a TaqMan real-time PCR method to detect the mutation of TaPIN1. • The distribution of Theileria annulata resistant to buparvaquone was investigated in Xinjiang, China. [ABSTRACT FROM AUTHOR]

Aim: A lucid, rapid and precise stability-indicating method was developed by using HPLC for the estimation of Buparvaquone in bulk as well as pharmaceutical dosage form by forced degradation studies. Materials and Methods: Princeton C18 column (4.6 × 150 mm, 5µ) and mobile phase containing 1% glacial acetic acid and acetonitrile in the proportion of 5:95 v/v was used throughout the study. The flow rate was 0.9 ml/m and the detecting wavelength was kept as 251 nm using the PDA detector. Results: The retention time of Buparvaquone was found to be at 8.6 ± 0.5 m. The method developed was validated, as stated in ICH Q2 (R1) guidelines. It was found to be linear within concentration ranging from 2-20 µg/ml having a correlation coefficient 0.999 and other parameters are also under permissible limits. Conclusion: Buparvaquone was exposed to different stress conditions like acidic, basic, neutral, thermal, peroxide and also photolytic. Amongst all, the drug was found to be more degraded under basic as well as peroxide conditions. [ABSTRACT FROM AUTHOR]

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Buparvaquone (BPQ)–loaded asymmetric solid lipid nanoparticles (SLN) prepared by a modified nanoprecipitation method were evaluated for splenotropic drug delivery. BPQ SLN exhibited an average particle size of 650.28 ± 6.75 nm with polydispersity index ≤ 0.3, entrapment efficiency of 96.57 ± 0.190%, and drug loading of 24.63 ± 0.042%. Scanning electron microscopy (SEM) revealed elongated particles with flattened and rounded edges. Aspect ratio, an important determinant of asymmetricity of the BPQ SLN, measured as the ratio of average length (1143 ± 0.083 nm) to width (419 ± 0.031 nm) was found to be 2.727 ± 0.19. The hemolytic potential of 10.86 ± 0.04% and good serum stability suggested feasibility for intravenous administration. 99mTc-labeled BPQ SLN revealed high radiolabeling efficiency (> 95%) and good stability. Intravenous administration in mice revealed > 75% accumulation in the reticuloendothelial system organs. The percent radioactivity per gram of organ was in the order spleen > kidney > lungs > liver > lymph nodes, with high splenic accumulation and significantly lower concentration in the liver. An astoundingly high spleen/liver ratio with a maximum of 11.94 ± 1.37 at 3 h, which confirmed high splenic uptake is attributed to Kupffer cell bypass. Other factors contributing to splenotropy are the rigidity and the low molecular weight of the lipid in the BPQ SLN which enabled translocation of the particles into the splenic pulp. Our study proposes asymmetric BPQ SLN as a promising splenotropic delivery system for improved efficacy in theileriosis, a spleen resident infection. [ABSTRACT FROM AUTHOR]

The study was carried out to determine clinical signs, hematological and therapeutic efficacy in T. equi infected horses. Clinical signs were pyrexia, anemia, icteric mucus membrane and fetlock swelling. Hematological examination revealed significantly decreased hemoglobin, packed cell volume, RBC and WBC count. The diseased horses were treated with Buparvaquone 4-6 mg/kg b.wt. intramuscularly along with hematinics and fluid therapy. [ABSTRACT FROM AUTHOR]

For the study 71 cattle calves were screened for theileriosis on the basis of classical clinical symptoms of theileriosis in calves. All screened calves underwent for PCR analysis, which revealed positive result in 12 calves. Clinical data of theses 12 calves were recorded separately and serum and blood samples of these 12 calves were investigated for haematology, serum biochemistry and bovine specific cardiac biomarkers i.e. cardiac troponin I and CPK-MB. Conventional therapeutic management with buparvoquone, oxytetracycline, and vitamin and iron supplementation was done in these 12 theileriosis affected calves and revaluation was done in recovered calves (8) after 15 days to assess the treatment response in terms of clinical recovery, haematobiochemical parameters and cardiac health evaluation through cardiac biomarkers. [ABSTRACT FROM AUTHOR]

Abstract Drug resistance is one of the emerging and re-emerging epidemics affecting both veterinary and public health sectors. Buparvaquone provides the most satisfactory means in the treatment of bovine tropical theileriosis. However, recently there has been widespread reports of development of resistance of Theileria annulata to buparvaquone. To investigate the situation in Sudan where bovine tropical theileriosis is endemic, fifty blood samples from T. annulata -positive cattle. were used for DNA extraction, PCR and cytochrome b gene nucleotide sequencing. Analysis of the two buparvaquone binding site regions Q 01 (130–148) and Q 02 (244–266), revealed three non- synonymous mutations at codon 146; alanine (GCT) to threonine (ACT) within the Q 01 region across all 50 isolates and the other mutation at codon 129; serine (AGC) to glycine (GGC) in 18 isolates which is very close to the Q 01 binding site. However, we documented another mutation at position 227; valine (GTG) to methionine (ATG) close to the close to the Q 02 binding site, in three isolates with mutation at codon 129. We concluded that this study has provided evidence of point mutations in the cytochrome b gene of T. annulata that might be associated with buparvaquone treatment failure in Sudan. [ABSTRACT FROM AUTHOR]

Aim: The aim was to determine hemato-biochemical alterations and to determine the better treatment of theileriosis in naturally infected calves. Materials and Methods: A total of 74 Holstein crossbred calves below 6 months of age, of either sex were included for present investigation in Bikaner. Based on the clinical examinations and laboratory results, 20 calves included for hemato-biochemical studies (before and after treatment) and divided into two groups (having 10 calves each). Results: The clinical examination of these calves revealed weakness, ticks infestations, high fever above 104°F, pronounced swelling of prescapular, prefemoral, parotid lymph nodes, loss of elasticity of skin, anemia, lacrimal discharges, pulpy cornea, tachycardia, and dyspnea. There was highly significant decrease (p

Buparvaquone is now the principle medicinal drug utilized against tropical theileriosis. The current study was conducted to determine the potential association between the resistance against buparvaquone and the occurrence of mutations in the Cytochrome b (Cyto b) gene of Theileria annulata (T. annulata) in cattle under Egyptian conditions. We sequenced the Cyto b gene of T. annulata of 6 blood samples collected from 1 responding and 5 non-responding cattle to buparvaquone treatment. The analysis revealed that two mutations occurred in codons 253 and 262 within the quinol oxidation (Qo2) drug-binding site of the five resistant isolates. Two mutations in the Qo1 site were found together in the buparvaquone sensitive isolate only. Our findings suggest that the failure of buparvaquone therapy could be attributed to the presence of single or double point mutations at the Qo2 site of T. annulata Cyto b gene. [ABSTRACT FROM AUTHOR]

The naphthoquinone buparvaquone is currently the only drug used against theileriosis. Here, the effects of buparvaquone were investigated in vitro and in an experimental mouse model for Neospora caninum infection. In 4-day proliferation assays, buparvaquone efficiently inhibited N. caninum tachyzoite replication (IC50 = 4.9 nM; IC100 = 100 nM). However, in the long term tachyzoites adapted and resumed proliferation in the presence of 100 nM buparvaquone after 20 days of cultivation. Parasiticidal activity was noted after 9 days of culture in 0.5 µM or 6 days in 1 µM buparvaquone. TEM of N. caninum infected fibroblasts treated with 1 µM buparvaquone showed that the drug acted rather slowly, and ultrastructural changes were evident only after 3–5 days of treatment, including severe alterations in the parasite cytoplasm, changes in the composition of the parasitophorous vacuole matrix and a diminished integrity of the vacuole membrane. Treatment of N. caninum infected mice with buparvaquone (100 mg/kg) either by intraperitoneal injection or gavage prevented neosporosis symptoms in 4 out of 6 mice in the intraperitoneally treated group, and in 6 out of 7 mice in the group receiving oral treatment. In the corresponding controls, all 6 mice injected intraperitoneally with corn oil alone died of acute neosporosis, and 4 out of 6 mice died in the orally treated control group. Assessment of infection intensities in the treatment groups showed that, compared to the drug treated groups, the controls showed a significantly higher parasite load in the lungs while cerebral parasite load was higher in the buparvaquone-treated groups. Thus, although buparvaquone did not eliminate the parasites infecting the CNS, the drug represents an interesting lead with the potential to eliminate, or at least diminish, fetal infection during pregnancy.

Abstract Canine theileriosis is a notorious tick borne piroplasmid infection of wild and domestic canines. The causative agent has not yet been accurately classified. PCR studies revealed that causative agent resembles to Theileria genus and thus provisionally named as Theileria annae. The other Theileria species reported in canines is Theileria annulata, Theileria equi and unnamed Theileria specie. This emergent canine infection is considered to be endemic in most of the European countries. However in Asia this disease has not been reported till date. The vectors responsible for transmission of this disease have not been determined. It has been suggested that DNA of Theileria annae has been detected in hard tick Ixodes hexagonus in Northwestern Spain and several other tick species. Clinically canine theileriosis is characterized by severe weakness, fever, hemoglobinuria and anemia. Recently atovaquone or buparvaquone plus azithromycin therapy showed better clinical efficacy. This comprehensive review is intended to summarize the current knowledge on prevalence and epidemiology of canine theileriosis in different countries of the world and associated tick vectors. Highlights • Canine theileriosis is a notorious tick borne piroplasmid infection. • The infection is considered endemic in Europe but not studied to this extent in Asia. • Responsible vectors for transmission of this disease have not been precisely determined. • The clinical picture of disease is quite severe. • Atovaquone or buparvaquone with azithromycin have shown clinical efficacy. [ABSTRACT FROM AUTHOR]

We here assessed the in vitro efficacy of the naptho-quinone buparvaquone (BPQ) against Besnoitia besnoiti tachyzoites in vitro. BPQ is currently licensed for the treatment of theileriosis in cattle in many countries, but not in the EU. In 4-day treatment assays, BPQ massively impaired tachyzoite proliferation with an IC50 of 10 ± 3 n m , and virtually complete inhibition was obtained in the presence of n m BPQ. Exposure to 1 µ m BPQ leads to ultrastructural changes affecting initially the mitochondrial matrix and the cristae. After 96 h, most parasites were largely distorted, filled with cytoplasmic amylopectin granules and vacuoles containing components of unknown composition. Host cell mitochondria did not appear to be notably affected by the drug. However, upon prolonged exposure (14–16 days) to increased BPQ concentrations, B. besnoiti tachyzoites exhibited the capacity to adapt, and they resumed proliferation at dosages of up to 10 µ m BPQ, albeit at a lower rate. These BPQ-adapted parasites maintained this lower susceptibility to BPQ treatment after freeze–thawing, and inspection by the transmission electron microscopy revealed that they underwent proliferation in the absence of structurally intact mitochondria. [ABSTRACT FROM AUTHOR]

Canine babesiosis is a tick-borne disease caused by several Babesia spp. which have different susceptebility to anti-protozoal drugs. A few drugs and drug combinations are used in the treatment of canine babesiosis often without complete parasite elimination leaving treated dogs as carriers which could relapse with clinical disease and also transmit infection further. Although the large form canine babesial species Babesia canis , Babesia vogeli and Babesia rossi are sensitive to the aromatic diamidines imidocarb dipropionate and diminazene aceturate, small form species such as Babesia gibsoni , Babesia conradae and Babesia vulpes ( Theileria annae ) are relatively resistant to these drugs and are treated with the combination of the hydroxynaphthoquinone atovaquone and the antibiotic azithromycin. Azithromycin and other antibiotics that have anti-protozoal properties target the apicoplast, a relict plastid found in protozoa, and exert a delayed death effect. The triple combination of clindamycin, diminazene aceturate and imidocarb dipropionate is also effective against B. gibsoni and used to treat atovaquone-resistant strains of this species. Novel drugs and the synergistic effects of drug combinations against Babesia infection should be explored further to find new treatments for canine babesiosis. [ABSTRACT FROM AUTHOR]

Background: Theileriosis is a tick-borne disease caused by Theileria strains of the protozoan species. Buparvaquone is the mostly preferred drug in the treatment theileriosis, while it is safety in sheep, has not been detailed investigated. It has been hypothesized that buparvaquone may show side effects and these effects may be defined some parameters measured from blood in sheep when it is used at the recommended dose and duration. The aim of this research was to determine the effect of buparvaquone on the blood oxidative status, cardiac, hepatic and renal damage and bone marrow function markers. Materials, Methods & Results: In this study, ten adult (> 2 years) Akkaraman rams were used. Healthy rams were placed in paddocks, provided water ad libitum, and fed with appropriate rations during the experiment. Buparvaquone was administered at the dose of 2.5 mg/kg (IM) intramuscularly twice at 3-day intervals. Blood samples were obtained before (0. h, Control) and after drug administration at 0.25, 0.5, 1, 2, 3, 4 and 5 days. The blood samples were transferred to gel tubes, and the sera were removed (2000 g, 15 min). During the study, the heart rate, respiratory rate, and body temperature were measured at each sampling time. In addition, the animals were clinically observed. Plasma oxidative status markers (Malondialdehyde, total antioxidant status, catalase, glutathione peroxidase, superoxide dismutase), serum cardiac (Troponin I, creatine kinase-MBmass, lactate dehydrogenase), hepatic (Alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase, total protein, albumin, globulin) and renal (Creatinine, blood urea nitrogen) damage markers and hemogram values (white blood cell, red blood cell, platelet, hemogram, hematocrit) were measured. Buparvaquone caused statistically significantly (P < 0.05) increases in the troponin I and blood urea nitrogen levels and fluctuations in alkaline phosphatase activity, but there was no any statistically significance difference determined in the other parameters. Discussion: In this study, buparvaquone was administered two times at a dose of 2.5 mg/kg (IM) at 3-day intervals. Although the result was not statistically significant (P > 0.05), it was determined that buparvaquone gradually increased the levels of the main oxidative stress marker, MDA, by approximately 2.8 fold. CAT and GPX levels were also found to have decreased by 2.2 fold. Buparvaquone may cause lipid peroxidation by producing free radicals. Some other antiprotozoal drugs may affect the oxidative status and may increase MDA level and decrease SOD level. In this study, MDA, which is an indicator of lipid peroxidation in vivo, was used to partially detect developing lipid peroxidation. Changes in the levels of reduced GPX and CAT enzymes could be attributed to their use in mediating the hydrogen peroxide detoxification mechanisms. The absence of significant changes in the TAS levels in this study suggests that buparvaquone may partially induce oxidative stress by producing hydrogen peroxide, but no significant changes occurred in the oxidative stress level because of the high antioxidant capacity of sheep. In this study, buparvaquone caused a statistically significant increase (P < 0.05) in the level of Tn-I, which is a marker of specific cardiac damage (P < 0.05), whereas there was no statistically (P > 0.05) significant increase in CK-MBmass. Tn-I and CK-MB levels, which are used to define heart damage in humans, have been successfully used to determine heart damage in sheep. In this research study, the statistically significant increases in Tn-I but not CK-MBmass levels could be considered indicative of mild cardiac damage. [ABSTRACT FROM AUTHOR]

A reliable and highly sensitive screening method based on liquid chromatography coupled with triple-quadrupole electrospray tandem mass spectrometry (LC-MS/MS) analysis has been developed for the detection and quantification of three veterinary drugs, including buparvaquone, nystatin, and etomidate impurity B CRS. The tested drugs were extracted from samples of porcine muscle, pasteurized whole milk, and eggs using 10 mM ammonium formate in acetonitrile followed by liquid-liquid purification with n -hexane. Chromatographic separation was achieved on a Phenomenex Luna C18 analytical column using 0.1% formic acid in ultrapure water (A) and acetonitrile (B) as mobile phases. All the matrix-matched calibration curves were linear ( R 2 ≥ 0.9756) over the concentration levels of the drugs tested. Recovery at two spiking levels (equivalent to the limit of quantification (LOQ) = 5 ng/g and 2 × LOQ) ranged from 72.88% to 92.59% with intra- and inter-day precisions <17%, except for porcine muscle spiked with 5 ng/g nystatin (RSD = 25.15%). Samples collected from markets located in Seoul, Republic of Korea, tested negative for all the drugs analyzed. In summary, this method is suitable for screening and quantifying the selected drugs in a single chromatographic run and with high selectivity in animal-derived food products meant for human consumption. [ABSTRACT FROM AUTHOR]

The present study was conducted to evaluate some effects of Buparvaquone on male fertility, as well as it's effects on the liver and kidney functions tests, some hematological parameters and histopathological changes. Therefore,75 mature male rats were used and equally divided into 5 groups; Group1, rats were injected with propylene glycol 2.5 ml/kg b.wt i.m and kept as a control group; Group2 rats were treated with Buparvaquone at a dose of 2.5 mg/kg b.wt i.m given as a single dose; Group3 rats were treated with Buparvaquone at a dose of 2.5 mg/kg b.wt i.m given twice with 2 weeks apart; Group4 rats were treated with buparvaquone at a dose of 5 mg/kg b.wt i.m given as a single dose. Group5 rats were treated with buparvaquone at a dose of 5 mg/kg b.wt i.m given twice with 2 weeks interval. Five rats from each treated and control groups were killed after 2, 4, 8 weeks from the beginning of drug administration. The obtained results showed that administration of buparvaquone at a dose of 5 mg/kg b.wt. i.m either injected once or twice induced a reduction in sperm cell count and packed cell volume at 4th and 8th week of the experiment. Also there was an elevation in Hb at 4th week and creatinine values at 8th week in the same groups. There was a significant increase in alkaline phosphate and creatinine values in group treated with buparvaquone at a dose of 5 mg/kg b.wt. i.m injected twice at 4th week. It could be concluded that buparvaquone is a safe drug when used with recommended dose. However; it induced some adverse effects in high repeated dose. [ABSTRACT FROM AUTHOR]