Theileria highjacks JNK2 into a complex with the macroschizont GPI (GlycosylPhosphatidylInositol)-anchored surface protein p104.

Constitutive c-Jun N-terminal kinase (JNK) activity characterizes bovine T and B cells infected with Theileria parva, and B cells and macrophages infected with Theileria annulata. Here, we show that T. annulata infection of macrophages manipulates JNK activation by recruiting JNK2 and not JNK1 to the parasite surface, whereas JNK1 is found predominantly in the host cell nucleus. At the parasite's surface, JNK2 forms a complex with p104, a GPI-(GlycosylPhosphatidylInositol)-anchor T. annulata plasma membrane protein. Sequestration of JNK2 depended on Protein Kinase-A (PKA)-mediated phosphorylation of a JNK-binding motif common to T. parva and a cell penetrating peptide harbouring the conserved p104 JNK-binding motif competitively ablated binding, whereupon liberated JNK2 became ubiquitinated and degraded. Cytosolic sequestration of JNK2 suppressed small mitochondrial ARF-mediated autophagy, whereas it sustained nuclear JNK1 levels, c-Jun phosphorylation, and matrigel traversal. Therefore, T. annulata sequestration of JNK2 contributes to both survival and dissemination of Theileria-transformed macrophages.

The Pathology of Pathogenic Theileriosis in African Wild Artiodactyls.

The published literature on schizont-"transforming," or pathogenic theileriosis, in African wild artiodactyls is dated and based on limited information. Here the authors review the taxonomy, diagnosis, epidemiology, hematology, pathology, and aspects of control in various species. Molecular studies based on 18S and 16S rRNA gene sequences have shown that African wild artiodactyls are commonly infected with diverse Theileria spp., as well as nontheilerial hemoprotozoa and rickettsia-like bacteria, and coinfections with pathogenic and nonpathogenic Theileria species are often recorded. Although theileriosis is still confusingly referred to as cytauxzoonosis in many species, the validity of a separate Cytauxzoon genus in artiodactyls is debated. The epidemiology of theileriosis is complex; the likelihood of fatal disease depends on the interplay of parasite, vertebrate host, tick vector, and environmental factors. Roan calves (Hippotragus equinus) and stressed animals of all host species are more susceptible to fatal theileriosis. Even though regenerative anemia is common, peripheral blood piroplasm parasitemia does not correlate with disease severity. Other than anemia, common macroscopic lesions include icterus, hemorrhages (mucosal, serosal, and tissue), fluid effusions into body cavities, lung edema, and variably sized raised cream-colored foci of leukocyte infiltration in multiple organs. Histopathologic findings include vasocentric hyperproliferation and lysis of atypical leukocytes with associated intracellular schizonts, parenchymal necrosis, hemorrhage, thromboembolism, and edema. Immunophenotyping is required to establish the identity of the schizont-transformed leukocytes in wild ungulates. Throughout the review, we propose avenues for future research by comparing existing knowledge on selected aspects of theileriosis in domestic livestock with that in African wild artiodactyls.

An appraisal of oriental theileriosis and the Theileria orientalis complex, with an emphasis on diagnosis and genetic characterisation.

Oriental theileriosis, a tick-borne disease of bovids caused by members of the Theileria orientalis complex, has a worldwide distribution. Globally, at least 11 distinct genotypes of T. orientalis complex, including type 1 (chitose), type 2 (ikeda), type 3 (buffeli), types 4 to 8, and N1-N3, have been described based on the sequence of the major piroplasm surface protein (MPSP) gene. Of these 11 genotypes, mainly ikeda and chitose are known to be pathogenic and cause considerable morbidity (including high fever, anaemia, jaundice and abortion), production losses and/or mortality in cattle. Mixed infections with two or more genotypes of T. orientalis is common, but do not always lead to a clinical disease, posing challenges in the diagnosis of asymptomatic or subclinical forms of oriental theileriosis. The diagnosis of oriental theileriosis is usually based on clinical signs, the detection of piroplasms of T. orientalis in blood smears, and/or the use of serological or molecular techniques. This paper reviews current methods used for the diagnosis of T. orientalis infections and the genetic characterisation of members of the T. orientalis complex, and proposes that advanced genomic tools should be established for investigations of these and related haemoparasites.

PKA and Apicomplexan Parasite Diseases.

The cAMP-dependent protein kinase PKA is a well-characterized member of the serine-threonine protein AGC kinase family and is the effector kinase of cAMP signaling. As such, PKA is involved in the control of a wide variety of cellular processes including metabolism, cell growth, gene expression and apoptosis. cAMP-dependent PKA signaling pathways play important roles during infection and virulence of various pathogens. Since fluxes in cAMP are involved in multiple intracellular functions, a variety of different pathological infectious processes can be affected by PKA signaling pathways. Here, we highlight some features of cAMP-PKA signaling that are relevant to Plasmodium falciparum-infection of erythrocytes and present an update on AKAP targeting of PKA in PGE2 signaling via EP4 in Theileria annulata-infection of leukocytes and discuss cAMP-PKA signling in Toxoplasma.

An outbreak of oriental theileriosis in dairy cattle imported to Vietnam from Australia.

This study reports an outbreak of oriental theileriosis in dairy cattle imported to Vietnam from Australia. Following clinical and pathological diagnoses, a total of 112 cattle blood samples were divided into three groups and tested using multiplexed tandem PCR. Group 1 were from aborted heifers in Vietnam; group 2 were from cattle before shipment from group 1 cattle and group 3 were from the same batch of cattle but transported to Taiwan. Theileria orientalis DNA was detected in 72·3% cattle. The prevalences of T. orientalis in groups 1, 2 and 3 were 77·6, 86·9 and 57·5%, respectively, and the difference in prevalence was significant between groups 1 and 3 (P < 0·0001). The infection intensities of genotypes chitose and ikeda of T. orientalis were higher in groups 1 (57 721 and 33 709, respectively) and 3 (5897 and 61 766, respectively) than those in group 2 (2071 and 6331, respectively). Phylogenetic analyses of the major piroplasm surface protein sequences revealed that genotypes chitose and ikeda determined herein were closely related to those previously reported from Australia. This first report of an outbreak of oriental theileriosis in imported cattle emphasizes improved measures for the export and import of cattle infected with T. orientalis.

Dermatological signs in bovine tropical theileriosis (T heileria annulata infection), a review.

Bovine tropical theileriosis (Theileria annulata infection) is an important tick-borne disease in several regions of the world. This paper describes two clinical cases of tropical theileriosis in northern Tunisia with the uncommon sign of skin nodules. The density of nodules was estimated at 15-20 per 10 cm2; they were distributed over the animal's body and were 0.5-2 cm in diameter. Microscopic examination of the skin nodules 18 days after treatment onset showed a perivascular infiltrate of lymphocytes, macrophages and eosinophils. Nevertheless, no schizonts were observed. Appropriate treatment led to the recovery of one of the two animals with total disappearance of the skin lesions; the other animal died. A review of clinical cases of tropical theileriosis (T. annulata infection) with cutaneous lesions is presented. Various dermatological signs are described in the literature: skin nodules, haemorrhagic lesions, cutaneous gangrene, etc. Most authors reported the presence of schizonts in the nodular lesions. Dermatological involvement in bovine tropical theileriosis should be considered by field veterinarians and should be differentiated from other diseases with dermatological signs.

La theilériose tropicale bovine (infection à Theileria annulata) est une maladie majeure à transmission vectorielle présente dans plusieurs régions du globe. Les auteurs décrivent deux cas cliniques de theilériose tropicale survenus dans le nord de la Tunisie, qui présentaient un tableau peu courant de lésions nodulaires. La densité estimée des nodules était de l'ordre de 15 à 20 nodules pour 10 cm² ; ils mesuraient de 0,5 à 2 cm de diamètre et étaient présents sur toutes les parties du corps de l'animal. L'examen au microscope des nodules cutanés 18 jours après le début du traitement a révélé une infiltration périvasculaire de lymphocytes, macrophages et éosinophiles. En revanche, aucun schizonte n'a été observé. L'administration d'un traitement approprié a éliminé l'infection chez l'un des deux animaux, avec une disparition totale de ses lésions cutanées ; l'autre animal est mort. Les auteurs présentent une synthèse des cas cliniques de theilériose tropicale (infection à T. annulata) s'accompagnant de lésions cutanées. Plusieurs manifestations dermatologiques sont décrites dans la littérature : nodules cutanés, lésions hémorragiques, gangrène cutanée, etc. La plupart des auteurs signalent la présence de schizontes dans les lésions nodulaires. Les manifestations dermatologiques associées à la theilériose tropicale bovine doivent être prises en compte par les vétérinaires de terrain et faire l'objet d'un diagnostic différentiel par rapport à d'autres maladies présentant un tableau dermatologique comparable.

La teileriosis tropical bovina (infección por Theileria annulata) es una importante enfermedad transmitida por garrapatas que afecta a varias regiones del mundo. Los autores describen dos casos clínicos de teileriosis tropical diagnosticados en el norte de Túnez que presentaban un síntoma inhabitual: nódulos cutáneos, con una densidad estimada de 15 a 20 nódulos por cada 10 cm2. Los nódulos, distribuidos por todo el cuerpo del animal, tenían un diámetro de entre 0,5 y 2 cm. El examen microscópico de los nódulos a los 18 días de tratamiento reveló una infiltración perivascular de linfocitos, macrófagos y eosinófilos, pero no se observaron esquizontes. La administración del tratamiento apropiado llevó a la recuperación de uno de los dos ejemplares, con desaparición completa de las lesiones cutáneas, mientras que el otro murió. Los autores pasan revista a una serie de casos clínicos de teileriosis tropical (infección por T. annulata) con presencia de lesiones cutáneas. En la bibliografía están descritos varios casos con síntomas dermatológicos: nódulos cutáneos, lesiones hemorrágicas, gangrena cutánea, etc. La mayoría de los autores señalan la presencia de esquizontes en las lesiones nodulares. Los veterinarios que trabajan sobre el terreno deben tener en cuenta que la teileriosis tropical bovina puede acompañarse de afectación cutánea y distinguir esta enfermedad de otras patologías que provocan síntomas dermatológicos.

The diagnostic importance of coagulation parameters in cattle having natural theileriosis.

The purpose of this study was to determine the diagnostic importance of coagulation parameters in cattle with natural theileriosis. Nine Holstein cross-breed cattle with theileriosis as infected group and 6 healthy Holstein cattle as control group were used in the present study. Mean fibrinogen level, thrombin time (TT), activated partial thromboplastin time (aPTT) and prothrombin time (PT) were not statistically different when control and infected groups compared, except for the D-dimer concentration. Quantitative D-dimer concentrations were determined by immune-turbidimetric assay. D-dimer values increased significantly (p<0.05) in infected group (631.55 ± 74.41 µg/L) compared to control group (370.00 ± 59.94 µg/L). D-dimer sensitivity and specificity were also determined at cut-off concentrations (372 µg/L). Sensitivity and specificity of D-dimer values were determined to be 88.89% and 83.33%, respectively. D-dimer is thought to be important indicator in the evaluation of the prognosis in theileriosis cases. Analysis of D-dimer values before and after treatment in controlled case studies were suggested in future studies to enlighten the issue.

Intracellular Theileria annulata promote invasive cell motility through kinase regulation of the host actin cytoskeleton.

The intracellular, protozoan Theileria species parasites are the only eukaryotes known to transform another eukaryotic cell. One consequence of this parasite-dependent transformation is the acquisition of motile and invasive properties of parasitized cells in vitro and their metastatic dissemination in the animal, which causes East Coast Fever (T. parva) or Tropical Theileriosis (T. annulata). These motile and invasive properties of infected host cells are enabled by parasite-dependent, poorly understood F-actin dynamics that control host cell membrane protrusions. Herein, we dissected functional and structural alterations that cause acquired motility and invasiveness of T. annulata-infected cells, to understand the molecular basis driving cell dissemination in Tropical Theileriosis. We found that chronic induction of TNFα by the parasite contributes to motility and invasiveness of parasitized host cells. We show that TNFα does so by specifically targeting expression and function of the host proto-oncogenic ser/thr kinase MAP4K4. Blocking either TNFα secretion or MAP4K4 expression dampens the formation of polar, F-actin-rich invasion structures and impairs cell motility in 3D. We identified the F-actin binding ERM family proteins as MAP4K4 downstream effectors in this process because TNFα-induced ERM activation and cell invasiveness are sensitive to MAP4K4 depletion. MAP4K4 expression in infected cells is induced by TNFα-JNK signalling and maintained by the inhibition of translational repression, whereby both effects are parasite dependent. Thus, parasite-induced TNFα promotes invasive motility of infected cells through the activation of MAP4K4, an evolutionary conserved kinase that controls cytoskeleton dynamics and cell motility. Hence, MAP4K4 couples inflammatory signaling to morphodynamic processes and cell motility, a process exploited by the intracellular Theileria parasite to increase its host cell's dissemination capabilities.

The aetiology, pathogenesis and control of theileriosis in domestic animals.

The Theileria genus includes a large number of species of tick-borne parasites that infect domestic animals and wildlife species, predominantly ruminants. These range from species, such as T. parva and T. annulata, which cause acute lymphoproliferative diseases in cattle resulting in high levels of mortality, to others that are non-pathogenic. In the last decade, several new pathogenic species of Theileria have been identified and pathogenic strains of other previously low-pathogenic species have emerged. Theileria parasites are characterised by developmental stages within leukocytes and erythrocytes. The capacity of the most pathogenic species to undergo extensive multiplication during intra-leukocyte development is central to their ability to cause disease. However, this is not the sole property responsible for disease, as illustrated by T. parva, which grows in a similar mannerin buffalo cells butdoes notcause disease inthisspecies. Because of the highly pathogenic nature of these parasites in livestock and the susceptibility of young animals to disease, control of the diseases is challenging. Control by chemotherapy and prevention of tick infestation has proved expensive and difficult to sustain. Vaccines using live parasites are available for T. parva and T. annulata and have been used with some success in the field. However, their widespread use has been hampered by practical constraints in production and distribution of the vaccines. Studies of the immune responses in immune cattle have helped to elucidate the protective immune responses and identified a number of parasite antigens that are currently being explored for development of alternative vaccines.

"Ormilo disease" a disorder of zebu cattle in Tanzania: bovine cerebral theileriosis or new protozoan disease?

"Ormilo" disease is a neurological disorder of cattle described by Maasai herders in Tanzania. It is attributed to infection by Theileria species, although no detailed data are available in the literature. The authors describe the macroscopical and histological changes observed in 30 brains of indigenous short-horn zebu cattle from Northern Tanzania, aged 2-9 years, with the characteristic neurological signs of "Ormilo". Moreover, the ultrastructural details observed in 14 selected brain samples were reported. Areas of congestion and hemorrhages, associated with the obstruction of the cerebral vessels with large numbers of parasitized lymphoid cells, were observed. Electron microscopy showed the presence of intralymphocytic parasites morphologically comparable to flagellated protozoa, not previously described in the lymphoid cells of cattle, but only reported during the sexual stages within the vector. Theileria taurotragi was detected by polymerase chain reaction (PCR) and reverse line blot (RLB) in nine samples. The authors hypothesize that the parasite detected by electron microscopy could be a strain of a Theileria endemic to this region till now not investigated, having an intralymphocytic phase and being associated with other Theileria spp. infestation. Further studies are needed to better understand the etiology of "Ormilo" disease and to characterize the morphology of the observed parasite, clarifying its role in the disease in Tanzania.

Clinicopathological findings of a natural outbreak of Theileria annulata in cattle: an emerging disease in southern Iran.

Theileriosis is an economically important hemoprotozoal disease with high morbidity and mortality in cattle. The present study reported the pathological features of a natural outbreak of tropical bovine theileriosis due to Theileria annulata in Fars Province, southern Iran. T. annulata was confirmed by the presence of T. annulata piroplasms in the blood smears and also by polymerase chain reaction test. On necropsy, pale mucous membranes and petechial and ecchymotic hemorrhages in the mucosal and serosal surfaces together with lymphadenopathy were observed. The liver was friable, yellowish, and larger than normal. Hemorrhages and punched-out ulcers were observed in the abomasal mucous membrane. Severe petechial hemorrhages were seen in the skin particularly in the hairless areas. Pulmonary edema and emphysema with petechial and ecchymotic hemorrhagic foci in the lungs were evident. The main histological changes were proliferation of lymphocytes in the lymph nodes and proliferation of macrophages, lymphocytes, and plasma cells in the spleen, Peyer's patches, portal tracts of the liver, and interstitial tissue of the kidneys. The mucous membrane of the abomasum showed numerous multifocal areas of necrosis and ulceration, and the submucosal area and lamina propria adjacent to these lesions showed hyperemia and hemorrhages, with mononuclear cell infiltration. The skin showed multifocal necrotic changes, petechial and ecchymotic hemorrhages, and chronic dermatitis. The schizonts of Theileria were evident in the cytoplasm of the lymphocytes and macrophages of the lymph nodes, spleen, and skin. Molecular examination revealed that these animals were infected with T. annulata. The present study describes the clinicopathological findings of bovine tropical theileriosis in an unpredictable weather condition.

Evaluation of erythrocyte antioxidant mechanisms: antioxidant enzymes, lipid peroxidation, and serum trace elements associated with progressive anemia in ovine malignant theileriosis.

Ovine malignant theileriosis is a fatal disease that is characterized by severe progressive anemia. In order to elucidate the underlying mechanisms involved in anemia, this study was designed to assess the antioxidant status and erythrocyte oxidative injuries in Iranian fat-tailed sheep that suffered from malignant theileriosis. The infected animals (infected group), composed of 50 Iranian sheep about 1-2 years old, naturally infected with Theileria sp., were divided into three subgroups according to parasitemia rates (<1%, 1-3%, 3-5%), and ten non-infected animals were also selected as the control group. Blood samples were taken and hematological parameters, the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase, erythrocyte osmotic fragility, and serum concentrations of some trace elements (copper, iron, zinc, manganese, and selenium), were measured. As an index of lipid peroxidation, the level of malondialdehyde (MDA) was also determined. According to the results, a significant decrease in red blood cell (RBC) count, packed cell volume, the activities of SOD, GPX, and catalase (P <0.001), and also serum concentrations of Cu, Zn, Mn, and Se (P < 0.05) were evident in the infected sheep. In contrast, significantly increased levels of MDA and erythrocyte osmotic fragility (P < 0.001) as well as serum concentration of iron (P < 0.05) were recorded in the infected animals. The significant decrease in antioxidant enzyme activities and substantial elevated levels of lipid peroxidation and erythrocyte osmotic fragility associated with the increase in parasitemia indicate increased exposure of RBCs to oxidative damage. Also, it appears that disturbed antioxidant defense mechanisms can promote the development of anemia in ovine theileriosis.

The role of sheep (Ovis aries) in maintaining Theileria orientalis Ikeda type infection.

Theileria orientalis is a tick‒borne intracellular parasite of red blood cells that causes severe and mild infections in various ruminants worldwide. To date there have been 11 types identified within this species, of which 4 types are presently found in New Zealand cattle. Since 2012, New Zealand has suffered a substantial epidemic of infectious bovine anaemia in both dairy and beef cattle associated with the Ikeda type. The speed at which the disease spread through the North Island suggested that other species could have been involved in transmission. The aim of a series of related experiments was to test the null hypothesis that sheep cannot maintain T. orientalis Ikeda type infection or infect ticks that feed on them. Several studies were conducted over 2 years to address this hypothesis which together showed that sheep can have detectable levels of T. orientalis Ikeda type infection in both the acute and chronic phase and that Haemaphysalis longicornis larvae can become infected when feeding on sheep. No anaemia, weight loss or clinical disease was recorded in the sheep in the acute phase of infection. The levels of infection recorded in the sheep were much lower than those found in cattle, consistent with the sheep being asymptomatic carriers of T. orientalis Ikeda type infection.

Effects of Theileria orientalis Ikeda type infection on libido and semen quality of bulls.

There is an epidemic in New Zealand of infectious bovine anaemia associated with Theileria orientalis Ikeda type, an obligate intracellular protozoan parasite. To establish whether T. orientalis Ikeda type infection adversely affects fertility of bulls used for natural mating, a randomised controlled experimental study was conducted. Ten of 17 2-year-old Friesian bulls that had not been previously infected with T. orientalis were infected with T. orientalis Ikeda type and then evaluations occurred during a 20-week period. There were semen and libido evaluations every 2 weeks, starting 4 weeks before the date of infection. In addition, there were blood collections, for haematocrit and infection intensity evaluations, rectal temperatures recorded, and bulls weighed three times weekly for 13 weeks after infection and then once weekly until completion of the study. Physical activity meters were also attached from Days 9-60 and 65-124 post-infection. The ten bulls were successfully infected with T. orientalis Ikeda type and this resulted in a decrease in HCT to about 0.25 by 70 days post-infection. There were no effects of infection on semen quality; however, during the acute phase of infection, when the infection intensity was rapidly increasing, the infected bulls took a longer time period for repeated mounting of females, and were less dominant in the herd social heiracrchy. In conclusion, although the transitory effects on libido could reduce conception rates, the overall effects of T. orientalis Ikeda type infection on bull fertility will probably be little.

Pathogenesis of liver lesions in Theileria orientalis-inoculated cattle and severe combined immunodeficiency mice with bovine erythrocyte transfusion.

Theileria orientalis (T. orientalis) is a bovine protozoal disease similar to malaria in humans. Although the common outcome of malaria in humans and T. orientalis infection in cattle is hepatic disorder, the mechanisms of its development remain unknown. In this study, we investigated hepatocyte injury characterized by accumulation of macrophages with ingested erythrocytes in sinusoid and extramedullary hematopoiesis in cattle and mice experimentally infected with T. orientalis (T. orientalis-infected cattle and T. orientalis-infected mice). Vacuolization of hepatic cells was frequently observed in the vicinity of the aggregated macrophages in the liver sinusoids of T. orientalis-infected mice. A significant percentage of the macrophages accumulated in the liver sinusoids of the severely infected cattle and mice (14.6% and 24.2 to 53.2%, respectively) reacted positively with interleukin-1, interleukin-6 and TNF-α antibodies. Increase in the production of these cytokines was confirmed in T. orientalis-infected cattle and mice by real-time RT-PCR. These findings strongly suggest that increased cytokine production by the macrophages that have phagocytosed T. orientalis-infected erythrocytes causes hepatic disorder in T. orientalis-infected animals.

Study on acute phase proteins (haptoglobin, serum amyloid A, fibrinogen, and ceruloplasmin) changes and their diagnostic values in bovine tropical theileriosis.

This study was conducted to assess the pattern of changes and the relative value of acute phase proteins (haptoglobin (Hp), serum amyloid A (SAA), ceruloplasmin and fibrinogen) in bovine tropical theileriosis. The diseased group comprised 50 dairy Holstein cattle, 2-3 years old, naturally infected with Theileria annulata. Infected animals were divided into four subgroups with different parasitemia rates (<1%, 1-3%, 3-5%, >5%). As a control group, 20 uninfected cattle were also sampled. Blood samples were collected and all measurements were done using validated methods. There were significant differences in red blood cells (RBCs), packed cell volume (PCV), hemoglobin (Hb) and concentrations of Hp, SAA, ceruloplasmin and fibrinogen between healthy cattle and those infected with T. annulata with different parasitemia rates (P < 0.05). As the parasitemia rate increased in infected cattle, a significant decrease was observed in RBCs, PCV, and Hb. In contrast, with increase in the parasitemia rate, a significant increase in the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), Hp, SAA, ceruloplasmin, and fibrinogen was evident. The optimal cut-off point was set by the receiver operating characteristics (ROC) method to >5.68 microg/ml for SAA, >0.16 g/l for Hp, >0.064 g/l for ceruloplasmin and >4.00 g/l for fibrinogen with corresponding 84% sensitivity and 100% specificity for SAA, 76% sensitivity and 100% specificity for Hp, 84% sensitivity and 80% specificity for ceruloplasmin and 40% sensitivity and 100% specificity for fibrinogen. SAA had the highest sensitivity and specificity and was diagnosed to be a suitable indicator of APPs changes in bovine theileriosis.

Infection dynamics of Theileria equi and Theileria haneyi, a newly discovered apicomplexan of the horse.

Theileria equi infection, exotic to the United States has reemerged through intravenous (iatrogenic) and tick-borne transmission. Surveillance at the US-Mexico border identified a new species, Theileria haneyi, (T. haneyiEP) (EP = Eagle Pass, Texas) which warranted additional investigation due to inability to detect by PCR targeting of T. equi ema-1 and EMA-1-cELISA validated for T. equi. Infection dynamics of T. haneyiEP were evaluated, including ability to superinfect in the presence of T. equi-Texas (T. equiTX), the isolate responsible for the reemergence of T. equi in the U S. Experimental infection with T. equiTX or T. haneyiEP revealed minimal clinical disease however, T. equiTX infection led to significantly greater neutropenia. Comparison of time to antibody detection following inoculation revealed significantly greater time to detectable anti-T. haneyiEP antibody (26.67 days post-inoculation (DPI)) than T. equiTX (11.67 DPI). Regardless of initial infection with either T. equiTX or T. haneyiEP, superinfection was established. Comparative analysis of antibody responses from a splenectomized horse infected with T. haneyiEP to that of a spleen intact horse infected with T. equiFL revealed a different antibody binding profile to T. haneyiEP, T. equiTX and T. equiFL merozoite antigen and limited shared antigen/cross-reactive antibody(s). Affinity purified T. equi EMA-1 and EMA-2 from T. equiFL were shown as targets for horse antibodies against T. haneyi. Data presented here show (1) T. haneyiEP can superinfect in the presence of T. equiTX infection and co-persists for minimally 25 months, (2) intravenous challenge with T. haneyi is subclinical, and (3) limited cross-reactive antibody between T. haneyiEP and T. equi includes reactivity to EMA-1 and EMA-2.

Effect of Peganum harmala (wild rue) extract on experimental ovine malignant theileriosis: pathological and parasitological findings.

Malignant theileriosis of sheep is a highly fatal, acute or subacute disease is caused by the tick-borne protozoan parasite, Theileria hirci. In this investigation ten healthy male lambs aged 5-6 months were randomly divided into two groups, A and B and were kept in isolated tick-proof pens. They were treated for internal and external parasite before commencement of the experiment. The lambs were experimentally infected with T. hirci by placing ticks Hyalomma anatolicum anatolicum infected with T. hirci on them. The ticks used in this survey had originally been isolated from sheep and colonies of them were established in an insectarium. Before and after infection rectal temperatures and clinical signs of the lambs were recorded, blood and prescapular lymph node smears were prepared and examined to determine the extent of the parasitaemia, and blood samples were analyzed to evaluate their haemoglobin (Hb) and packed cell volume (PCV) rates. Three days after the commencement of a febrile reaction and appearance of the schizonts in the lymph node smears, treatment of the lambs in Group A with an extract containing the alkaloids of Peganum harmala (wild rue) was commenced. Group B lambs were kept untreated controls. Before treatment there were no significant differences in the rectal temperature, parasitaemia rate, and the Hb and PCV values between animals in the two groups but after treatment significant differences in these values was detected (P < 0.05). After treatment, the clinical signs and parasites in the lymph node smears of the animals in Group A disappeared and they all animals recovered. These parameters in the animals of Group B progressed until their death. Pathological studies showed the characteristic lesions of theileriosis in lambs in Group B, but not in Group A. The results indicate a therapeutic effect of the alkaloids of P. harmala for treatment of ovine malignant theileriosis.

c-Jun NH2-terminal kinase/c-Jun signaling promotes survival and metastasis of B lymphocytes transformed by Theileria.

Theileria parasites infect and transform bovine lymphocytes resulting in tumors with metastatic/invasive potential. Importantly, cellular transformation is reversed upon drug-induced parasite death, and the infected lymphocyte dies of apoptosis within 48 hours. Theileria-dependent transformation leads to the constitutive activation of c-Jun NH2-terminal kinase (both JNK1 and JNK2) and permanent induction of activator protein-1. Inactivation of JNK (following transfection of dominant-negative mutants, or treatment with a JNK-specific inhibitor) leads to lymphocyte apoptosis, suggesting an antiapoptotic role for JNK activation in Theileria-induced B cell transformation. Theileria-induced JNK activation also leads to constitutive c-Jun phosphorylation, and inhibition of c-Jun and activator protein-1 transactivation following the expression of a dominant-negative mutant of c-Jun sensitizes Theileria-transformed B cells to apoptosis, but does not significantly affect their proliferation. Thus, JNK activation and c-Jun induction have overlapping, but nonidentical antiapoptotic roles in Theileria-induced B cell transformation. Increased sensitivity to apoptosis may be related to the fact that the expression levels of antiapoptotic proteins such as Mcl-1 and c-IAP are reduced upon c-Jun inhibition. In addition, decreased c-Jun expression correlates with the impaired ability of transfected B cells to degrade synthetic matrix in vitro, and their injection into lymphoid mice gives rise to significantly less and smaller tumors. Combined, these data argue for a role for JNK and c-Jun induction in the survival and metastasis of Theileria-transformed B cells. The similarity between Theileria-transformed B cells with human B lymphomas argues that exploiting the reversible nature of Theileria-induced transformation could throw light on the mechanisms underlying human malignancies.

Experimental infection of Friesian bulls with Theileria orientalis (Ikeda) and effects on the haematocrit, live weight, rectal temperature and activity.

Since 2012, New Zealand has suffered from an epidemic of infectious bovine anaemia associated with T. orientalis (Ikeda), an obligate intracellular protozoan parasite of cattle. Despite widespread agreement that T. orientalis (Ikeda) infection has impacted beef and dairy farming in New Zealand there is very little quantitative data to support this conclusion. A randomised controlled experimental study of the effect of T. orientalis (Ikeda) infection on the live weight, haematocrit (HCT), temperature and activity of 2-year-old Friesian bulls was conducted at a Massey University Research farm, Palmerston North. Ten out of seventeen 2-year-old Friesian bulls were injected intravenously with 30 mL whole blood from 2 clinical cases of Theileria-associated bovine anaemia and then followed over a period of 20 weeks. The bulls were blood sampled, had rectal temperature recorded and weighed 3 times weekly for 13 weeks and then once weekly thereafter until the end of the trial. Infection intensity was monitored using qPCR. All 10 inoculated bulls were successfully infected with T. orientalis (Ikeda). The results showed that the live weight response to infection was varied and the bulls could be divided into two groups based on this response. Four infected bulls showed a significant weight reduction of 41.5 kg (p < 0.0001), a financial loss of around NZ$112 per bull, compared with the other 6 bulls in the infected group, which were not different to the 7 uninfected controls. The live weight of the 4 poor growing bulls was significantly lower than the other 6 infected bulls from Day 71 post infection (p < 0.05). All ten infected bulls showed a similar decrease in HCT, with the lowest HCT reached around Day 60 to 80 post-infection, however the four infected bulls that grew poorly did have a significantly elevated HCT for the first 1 to 3 weeks post infection (p < 0.05). The 4 infected bulls which grew poorly also had a significantly higher infection intensity than the other infected bulls from Day 27 to Day 60 post-infection (p < 0.05). There was no pyrexia recorded in the infected group or control groups, instead there was a tendency for the infected group to have a lower rectal temperature from Day 5 to 70 post infection. The infected bulls walked on average 239 steps per day less than the control bulls, although this difference was not significant (p = 0.35). Overall the study clearly showed, by controlling infection date and infectious dose, that a proportion of cattle infected with T. orientalis (Ikeda) have significantly decreased live weight gains.