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Piroplasm including Babesia spp. and Theileria spp. in cattle can cause illness that affects livestock productivity, resulting in significant production losses, especially in tropical and subtropical regions such as Thailand. This study aimed to investigate the prevalence of bovine piroplasms and to identify these blood parasites based on the 18S ribosomal RNA gene in cattle in the northeastern part of Thailand. Piroplasmid infections among beef and dairy cattle were examined using nested PCR. Furthermore, amplicon DNA was sequenced and analyzed, and a phylogenetic tree was constructed to determine the genetic diversity and relationships of the parasite in each area. A total of 141 out of 215 (65.6%) cattle were positive for infection with Babesia or Theileria. DNA analysis revealed that infection by Babesia bigemina, Babesia bovis, Theileria orientalis, Theileria sinensis, and Theileria sp. were common piroplasms in cattle in this region, with a high sequence shared identity and similarity with each other and clustered with isolates from other countries. This study provides information on the molecular epidemiology and genetic identification of Babesia spp. and Theileria spp. in beef and dairy cattle to provide a better understanding of piroplasm infection in cattle in this region, which will help control these blood parasites. Moreover, this is the first report identifying T. sinensis circulating among Thai cattle., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Theileriosis is a tick-borne protozoal disease caused by a piroplasm of the genus Theileria. Hard ticks are obligate hematophagous ectoparasites that serve as vectors of Theileria spp. Studies of the presence of theileriosis in Egyptian dogs and associated ticks are scarce. This study was conducted to detect and identify Theileria spp. in dogs and Rhipicephalus sanguineus ticks and to monitor the epidemiological data of this disease. The prevalence rates of Theileria equi infection were 12.02%, 0.73%, 2.93%, and 1.83% by microscopic examination of dog blood, tick hemolymph, tick midgut, and tick salivary smears, respectively. Conversely, the T. equi prevalence in dog blood and associated ticks assessed by PCR was 25.81% and 10.42%, respectively. Epidemiological data about Theileria infection revealed a significant difference in the infection between different seasons and different dog breeds (p value <0.05), whereas host, sex, and age of dogs had no significant effect on the infection. Sequencing of PCR products showed that all PCR positive samples were infected with T. equi. Transmission electron microscopy (TEM) described the different stages of Theileria in the midgut and salivary gland of infected ticks. The current study confirmed that T. equi is not specific to equine hosts, and confirmed that dogs are a susceptible host to T. equi., (© 2022 Royal Entomological Society.)

Theileriosis is a tick-borne disease that causes enormous losses in the dairy industry. There are several species of Theileria that can infect bovines. Generally, more than one species are prevalent in any geographical area; thus, chances of co-infections are high. Differentiation of these species may not be possible by microscopic examination or serological tests. Therefore, in this study, a multiplex PCR assay was standardized and evaluated for rapid and simultaneous differential detection of two species of Theileria viz., Theileria annulata and Theileria orientalis. Species-specific primers were designed to target the merozoite piroplasm surface antigen gene (TAMS1) of T. annulata and the major piroplasm surface protein gene of T. orientalis, yielding specific amplicon of 229 bp and 466 bp, respectively. The sensitivity of multiplex PCR was 10 2 and 10 3 copies for T. annulata and T. orientalis, respectively. The simplex and multiplex PCRs were specific and showed no cross-reactivity with other hemoprotozoa for either primer. For comparative evaluation, blood samples from 216 cattle were tested by simplex and multiplex PCR for both species. Using multiplex PCR, 131 animals were found infected for theileriosis, of which 112 were infected with T. annulata, five were infected with T. orientalis, and 14 had mixed infections. This is the first report of T. orientalis from Haryana, India. Representative sequences of T. annulata (ON248941) and T. orientalis (ON248942) were submitted in GenBank. The standardized multiplex PCR assay used in this study was specific, sensitive, for the screening of field samples., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Theileriosis is one of the most important tick-borne diseases that has been affecting farmers and thousands of livestock in Zimbabwe. The main government strategy to combat theileriosis is use of plunge dips with anti-tick chemicals at specified times; however, an increase in number of farmers caused a strain on government services resulting in disease outbreak. One of the key issues that have been highlighted by department of veterinary is the strain in communication and knowledge of the disease with the farmers. Hence, it is important to evaluate the communication between farmers and veterinary services and identify possible areas of strain. A field survey was conducted with 320 farmers in Mhondoro Ngezi, a district badly affected by theileriosis. Face-to-face interviews with smallholders and communal farmers were conducted between September and October 2021, and the data were analyzed using Stata 17. Communal farmers relied mainly on oral communication and had limited knowledge of theileriosis; therefore, dead cattle % was high among them. Though veterinary extension officers were the prime source of information, oral communication medium affected knowledge transferred. The results of this study recommend adoption of communication mediums that encourage retention, such as brochures and posters by veterinary extension services. The government may also partner with private players to ease pressure of increased farming population due to land reform., (© 2023. The Author(s).)

Theileriosis is a hemoparasitic disease that affects a wide range of different animal species and is caused by various species of Theileria. This study aimed to determine the molecular epidemiology of Theileria annulata through microscopy and PCR, in crossbred cattle in some districts of Khyber Pakhtunkhwa, Pakistan. For this study, a total of 384 blood samples were collected from cattle in the Peshawar (n=120), Charsadda (n=94), Nowshera (n=84), and Swabi (n=86) districts. Microscopy and PCR were used to determine the overall prevalence of theileriosis, which was found to be 15.8 and 22.6%, respectively. Theileria annulata was detected in blood samples through PCR in the study area, and the target gene i.e., Tams 1, of positive samples was sequenced. The sequences in the current study revealed high sequence homology (ranging from 96 to 100%) with Tams 1 sequences of neighboring countries present in the NCBI database. Season, breed, age, and sex were found to be important risk factors among the several risk factors examined, whereas, among different clinical manifestations, lymphadenopathy showed a strong association with theileriosis. According to Cohen's kappa and ROC analysis, microscopy was proven to be a fair diagnostic test for detecting theileriosis in cattle, and may be used in combination with molecular techniques for screening a large number of animals., (Copyright© by the Polish Academy of Sciences.)

Theileriosis is one of the top ten economically important diseases in cattle in India. Cytokines are considered important mediators and regulators of the immune response to an infection. In the present study, the gene expression profiles of fourteen cytokines (IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL12A, IL12B, IL16, TGFB1, TNFA, IFNA and IFNB) were compared in Theileria annulata-infected and healthy crossbred cattle. Blood samples were obtained from the District Disease Diagnostic Laboratory, Karnal. The presence/absence of T. annulata infection in the animals was determined on the basis of blood smear examination and molecular detection through PCR using the genus-specific primers. Total RNA was extracted from peripheral blood mononuclear cells, which was further reverse transcribed to cDNA. Primer3 software was employed to design the primers for Real-Time qPCR. The results were examined using 2 -∆∆Ct  method with RPS15 and GAPDH as the reference genes. The expression of IL1B, IL6, IL8, IL10, IL12A, IL12B, TNFA, IFNA and IFNB was significantly higher, whereas the expression of IL2 was lower in the infected animals. The transcript levels of IL1A and TGFB1 were also higher in the diseased animals, but the results were non-significant. This study profiles the expression kinetics of various pro-inflammatory and anti-inflammatory cytokine genes in response to bovine theileriosis., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Ticks are economically important obligatory blood feeding arthropods that have a pivotal role in transmission of infection. The present study was conducted in ixodid ticks collected from four districts of coastal Odisha, India to investigate the prevalence of Theileria annulata . Adult semi engorged Hyalomma anatolicum ticks ( n  = 178) were dissected, the salivary gland was isolated and DNA was extracted. A nested PCR targeting the Tams1 gene of T. annulata , utilizing two sets of primers (N516F, N517R, and Ta14136iF, Ta249R) was utilized for detection of the parasite. The PCR products were then sequenced and subjected to BLAST analysis, alignment, and phylogenetic study. Two sequences deposited in GenBank were assigned Accession No MH477290.1 and Accession No MH477291.1. The molecular investigation of T. annulata revealed an overall prevalence of 14.6% in tick vectors, and nested PCR was found to have significant ( p  < 0.05) higher results than primary PCR. A significant higher presence ( p  < 0.05) was recorded in female ticks compared with male ticks. This is the first report of detection of the parasite in tick vectors in the state of Odisha.

Theileria parasites commonly infect African wild artiodactyls. In rare roan ( Hippotragus equinus ) and sable ( H. niger ) antelopes, Theileria sp. (sable)-associated calf mortalities constrain breeding programs. The pathogenicity of most leukocyte-transforming Theileria spp. originates in their invasion of and multiplication in various mononuclear leukocytes, the transformation of both infected and uninfected leukocytes, and their infiltration of multiple organs. Understanding the pathogenesis of theileriosis can be improved by the use of immunohistochemistry (IHC) to identify the localization of the parasites in tissue sections. Our aim was to develop a reproducible IHC assay to detect leukocyte-associated Theileria parasites in formalin-fixed, paraffin-embedded roan and sable tissues. Polyclonal antibodies were purified from the sera of 5 roans from an area endemic for Theileria sp. (sable) and tested for IHC reactivity in 55 infected and 39 control roan and sable antelopes, and for antigen and species cross-reactivity in an additional 58 cases. The 3 strongest antibodies consistently detected intraleukocytic theilerial antigens in known positive cases in roan and sable antelopes, and also detected other Theileria spp. in non-hippotraginid wild artiodactyl tissues. The antibodies did not cross-react with other apicomplexan protozoa, with the exception of Cryptosporidium . Given that PCR on its own cannot determine the significance of theilerial infection in wild ruminants, IHC is a useful laboratory test with which to confirm the diagnosis in these species.

This study aimed to investigate whether the infection intensity of Theileria orientalis Ikeda type organisms within Haemaphysalis longicornis larvae and nymph stages fluctuated over 6 mo after feeding as larvae on infected calves in the field. Naïve larvae, hatched from eggs, were fed on infected calves for 5 days while contained within cotton socks glued over the calves' ears. Larvae were first sampled immediately post-feeding and then sampled every 3 wk for 23 wk in total, after molting to nymphs. All larvae and nymphs were tested for T. orientalis Ikeda organisms using quantitative PCR. The qPCR results showed that the infection intensity of Haemaphysalis longicornis larvae and nymphs was not constant over the sampling period, and after initially dropping after molting to nymphs, it then rose with fasting to a maximum at 17 and 23 wk post-feeding. The significant rise in T. orientalis Ikeda organisms observed at 23 wk postfeeding may explain why more severe clinical cases of bovine theileriosis in New Zealand are seen in the spring when nymphs are the predominant instar questing., (© American Society of Parasitologists 2021.)

Cattle piroplasmoses are tick-borne diseases, spread worldwide that cause significant economic losses. A participatory epidemiological study was conducted individually or in focus groups with 73 cattle owners in Beni Hamidene locality (district of Constantine, Algeria). The aim of this study was to study cattle owners' knowledge, attitude and perception on cattle piroplasmosis. Proportional piling technique was used to determinate most common cattle diseases, and to evaluate economic impact of diseases according to the interwieved farmers. Theileriosis (49/73; 67.1%) and babesiosis (44/73; 60.3%) were considered the most important bovine diseases. No zoonotic disease was cited by the interviewed cattle owners. According to the majority of cattle owners, theileriosis and babesiosis are deadly diseases (87.3 and 78.1%, respectively). All cattle owners (73/73) cited fever as the most common symptom of tropical theileriosis. Some of them (14/73; 19.2%) do not make distinction between theileriosis and babesiosis. According to cattle owners (65/73; 89.0%), the use of acaricide is the most appropriate tick control method. This study provides information about knowledge on bovine piroplasmoses in Algeria. These information could be considered when performing control programmes by both animal decision-makers and field veterinarians.

The aim of this study was to test the hypothesis that toltrazuril administered at 4 weeks post-turnout reduces the infection intensity of Theileria orientalis Ikeda type in dairy calves and so prevents serious clinical disease in these animals at 2-3 months of age. Two groups of 40 dairy calves on two separate dairy farms in the Waikato were followed for 16 weeks post-turnout onto pasture. On each farm, 20 calves were randomly selected and orally treated with toltrazuril (15 mg/kg) at 4 weeks post-turnout, whilst the remaining 20 calves were left untreated. All 40 calves were blood sampled and weighed at 2, 4, 6, 8, 12, and 16 weeks post-turnout i.e. 6 samplings per calf. A random subset of 10 calves from each treatment group on each farm were faecal sampled at each visit. The blood samples were used to estimate the T. orientalis Ikeda type infection intensity and haematocrit for each calf and the faecal samples were used to estimate the number of coccidia oocysts per gram of faeces. Three linear mixed effects models, to evaluate the effect of toltrazuril treatment on infection intensity, haematocrit (HCT) and weight respectively were fitted to the data. No calves on either farm developed clinical theileriosis or coccidiosis and the three mixed effects linear models, controlling for the effect of farm and days from turnout, showed that there was no effect of treatment on infection intensity (p = 0.81), on HCT (p = 0.99) and on weight gain (p = 0.79). In conclusion, this study showed no evidence supporting the use of toltrazuril to control T. orientalis Ikeda type infection levels and prevent disease., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Bovine tropical theileriosis caused by Theileria annulata is an overwhelming haemoprotozoan tick-borne disease in taurine and cross-bred cattle in Punjab, India. However, there seems to be no report from India of cutaneous nodules associated with the disease. This report describes a five-year-old cross-bred cow presented to a university clinic with a history of fever, inappetence and malaise for the past six to seven days. Clinical examination revealed normal vital parameters, pale mucous membranes, mild enlargement of the prescapular lymph nodes and multiple subcutaneous nodular masses (2-4 cm) on the neck and abdomen. Haematology revealed mild anaemia and leucopenia with 48% neutrophils, 48% lymphocytes and 4% eosinophils. Romanowsky-stained smears of fineneedle aspiration biopsy samples from swollen lymph nodes and subcutaneous masses showed an increased number of lymphoid cells, suggesting cutaneous lymphomatosis. However, a critical examination of the smears from subcutaneous nodules showed a large number of Koch's blue bodies in macrophages and lymphoblasts, and several piroplasms were also noticed within the red blood cells in lymph node smears. A peripheral blood smear revealed mild to moderate parasitaemia. Extracted DNA from the parasitologically positive blood sample was subjected to nested polymerase chain reaction (nPCR) using T. annulata speciesspecific primers encoding the 30-kiloDalton major sporozoite surface antigen. The desired 572-base pair amplified product of the nPCR was comparable to the positive control. This seems to be a rare case of T. annulata in an adult cross-bred cow, showing cutaneous nodular involvement.

Since 1982 there have been two epidemics of bovine anaemia associated with Theileria orientalis infection (TABA) in New Zealand. The latest and more serious epidemic started in 2012 and is associated with the T. orientalis (Ikeda) type. In 2013 the Ministry for Primary Industries (MPI) carried out several epidemiological investigations on the TABA epidemic, which included a series of on-farm herd prevalence studies. The aims of this paper are to present and summarise the results of these herd prevalence studies. The data were collected from March 2013 to September 2013 and comprised 26 farms. For 21/26 of these farms, a T. orientalis (Ikeda) positive index case of TABA precipitated the follow-up herd prevalence study. Each herd prevalence study involved a random sample of 10 cows for haematocrit, for piroplasm counts (number of Theileria infected RBCs per 1000 RBCs) and for PCR molecular diagnosis. Animals were diagnosed anaemic if the haematocrit ≤0.24. The results showed that for the index cases, the average haematocrit = 0.10 L/L and the average piroplasm count = 10.9 Theileria infected RBCs per 1000 RBCs, and for the herd prevalence studies the average haematocrit = 0.26 and the average piroplasm count = 6.9. The average herd prevalence of T. orientalis (Ikeda) PCR positive cattle and of anaemic cattle was 87% and 26% respectively. For dairy herds (n = 7 herds) it took, on average, 80 days for the within herd prevalence of anaemia to return to zero after the initial diagnosis. In conclusion the prevalence studies showed that infection rates and anaemia rates were already high in herds when the index case of TABA was observed. At the time, these prevalence studies significantly added to our understanding of the epidemiology of the evolving T. orientalis (Ikeda) epidemic and furthermore met the New Zealand MPI objective of being rapid and applied, fit for purpose and completed at a relatively low cost., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Commencing in 2012, an epidemic of infectious bovine anaemia associated with Theileria orientalis Ikeda type has been present in New Zealand. The aims of this study were to analyse the temporal and spatial effects of T. orientalis Ikeda type infection on the sample submission rates and haematocrits of infected cattle over the first two years of the New Zealand epidemic. The data were collected from 30/08/2012 to 28/11/2014 and included all samples that met the case definition for Theileria associated bovine anaemia (TABA) and tested positive for T. orientalis Ikeda type by PCR. The sample submission rates by month and farm type were highly seasonal with dairy farm submissions peaking in September a month before beef farm submissions peaked. A second lesser peak of dairy farm submissions in April was absent for beef farms. A mixed effects model was fitted to the data and showed a significant interaction between farm production type (dairy or beef) and month of sampling (p=0.006) and between latitude and month of sampling (p=0.024). The estimated haematocrit, adjusted for month and latitude, for dairy cattle=0.125 (95%CI 0.121-0.129) and for beef cattle=0.151 (95% CI 0.138-0.165), p<0.0001. This research shows that infected beef animals tend to be less severely affected than dairy animals and that the month of sampling and latitude of the sampled farm have significant and interacting effects on the level of anaemia associated with T. orientalis Ikeda type infection., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The aim of the study was to observe changes in haematocrit (HCT) over time in a New Zealand South Island dairy herd affected by an outbreak of Theileria-associated bovine anaemia (TABA; Ikeda). A secondary aim was to relate individual cow HCTs to the amount of Theileria orientalis Ikeda DNA present in the blood, as measured by cycle threshold values, using a quantitative PCR (qPCR). Over a 6 month period, blood samples from 19 randomly selected cattle were monitored from a herd of 600 dairy cows. The sampling interval was approximately fortnightly for the first six weeks, followed by sampling at between four and six week intervals. At the initial report of the outbreak, two from six cattle were anaemic (HCT<0.25L/L). Blood collected from 14 cattle 11 days later showed that 57% (95% CI 33-77%) of the cattle sampled were anaemic. Of the 19 cattle that went on to be monitored, 12 (63% 95% CI=41-81%) developed anaemia at some point during the period of monitoring. One of the anaemic animals did not meet the case definition for TABA Ikeda. For individual cattle, the average number of days between when cattle were first detected as anaemic and when HCT returned to normal was 53days (median=47 days, range=6-92 days). At the point of notification the amount of T. orientalis Ikeda DNA in the blood of the six cattle tested was low (Cq median=36), but 11days later the amount of DNA in blood of 14 additional cows tested was relatively high (Cq median=24). Levels of all 19 cows monitored continued to remain moderately high through the period of testing (Cq median=29). This was despite a general improvement in the HCT of affected cattle. In four of the 15 cattle positive to T. orientalis Ikeda where blood fractions (plasma and whole blood) were tested, it appeared that T. orientalis Ikeda (as measured by qPCR) dropped more rapidly in plasma fractions than in whole blood at the point that HCT started to return to normal levels. Despite the assumption that tick populations were low in the Canterbury region of the South Island the impact of TABA (proportion of herd affected and the average period that animals remained anaemic) on the case herd was still relatively high., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objectives: To examine the prevalence of Theileria orientalis in the Gloucester area of New South Wales and its effect on individual animals., Methods: Blood samples (EDTA and clotted blood) were collected from a total of 55 calves and their dams from 6 properties over a 16-week period. A total of 202 and 190 blood samples were collected from the calves and dams, respectively, and were examined via blood film for the presence of intraerythrocytic T. orientalis piroplasms. Packed cell volume (PCV) was measured to determine infection resulting in anaemia. The presence of antibodies against the T. orientalis major piroplasm surface protein (MPSP) was tested using ELISA., Results: The overall prevalence of T. orientalis infection in both dams and calves from all herds examined was 95%. Mean peak parasitaemia was observed in calves between 6 and 9 weeks of age (P = 0.051), coinciding with a decline in mean PCV. Only 3 (6%) of the blood samples collected from the dams were positive for Theileria-associated antibodies and no significant relationship (P > 0.05) was found between the presence of antibodies in the dams and PCV levels in the calves. There was no evidence that passive transfer of antibodies from dams to calves protected the calves against a decline in PCV., Conclusion: This study confirmed a high prevalence of low-level Theileria infection, but low MPSP seroconversion rates, in dams and calves in an area where the disease has been endemic for a number of years., (© 2017 Australian Veterinary Association.)

Background: Bovine theileriosis, caused by the haemoprotozoan Theileria orientalis, is an emerging disease in East Asia and Australasia. Previous studies have demonstrated transplacental transmission of various Theileria spp. but molecular confirmation of transplacental transmission of T. orientalis has never been confirmed in the field. In this study, cow-calf (< 48 h old) pairs were sampled across 3 herds; opportunistic samples from aborted foetuses or stillborn calves were also examined. Molecular (multiplex qPCR) and serological (ELISA) methods were used to determine infection prevalence and the presence of anti-Theileria antibodies in each herd. In addition, pregnant heifers and foetal calves were sampled at abattoir and tested for the presence of T. orientalis by qPCR., Results: The qPCR results indicated that, even though there was a high prevalence of T. orientalis infection in cows, the rate of transplacental transmission to their calves was low, with only one newborn calf from one herd and one foetus from the abattoir testing positive for T. orientalis DNA. Five aborted foetuses and stillborn calves, 3 of which were derived from a herd experiencing a high number of clinical theileriosis cases at the time of sampling, all tested negative for T. orientalis by qPCR. This suggests that in utero infection of calves with T. orientalis may not be a major driver of abortions during theileriosis outbreaks. Temporal monitoring of 20 calves born to T. orientalis-positive mothers indicated that T. orientalis was detectable in most calves between 10 and 27 days post-partum, consistent with prior field studies on adult cattle introduced to Theileria-affected herds. There was a positive correlation between the ELISA ratio of newborn calves and their mothers within 48 h of calving; however, maternal antibodies were only detectable in some calves and only for 4-4.5 weeks post-partum. All calves displayed high parasite loads peaking at 4-8 weeks post-partum, with only some calves subsequently mounting a detectable adaptive antibody response., Conclusions: These findings indicate transplacental transmission of T. orientalis appears to play only a minor role in persistence of T. orientalis infection in the field; however calves are highly susceptible to developing high level T. orientalis infections at 4-8 weeks of age regardless of whether maternal antibodies are present post-partum.

Background: Bovine theileriosis is a common disease transmitted by ticks, and can cause loss of beef and dairy cattle worldwide. Here, an indirect enzyme-linked immunosorbent assay (iELISA) based on Theileria luwenshuni surface protein (TlSP) was developed and used to carry out a seroepidemiological survey of bovine theileriosis in northern China., Methods: We used the BugBuster Ni-NTA His•Bind Purification Kit to purify recombinant TlSP (rTlSP), which was subsequently analyzed by Western Blotting to evaluate cross-reactivity with other pathogen-positive sera. The iELISA method based on rTlSP was successfully developed. Sera from 2005 blood samples were tested with the rTlSP-iELISA method, and blood smears from these samples were observed by microscopy., Results: The specificity of iELISA was 98.9%, the sensitivity was 98.5%, and the cut-off was selected as 24.6%. Western Blot analysis of rTlSP confirmed that there were cross-reactions with Theileria luwenshuni, Theileria uilenbergi, Theileria ovis, Theileria annulata, Theileria orientalis and Theileria sinensis. The epidemiological survey showed that the highest positive rate of bovine theileriosis was 98.3%, the lowest rate was 84.1%, and the average positive rate was 95.4% by iELISA. With microscopy, the highest positive rate was 38.9%, the lowest rate was 5.1%, and the relative average positive rate was 13.7%., Conclusions: An rTlSP-iELISA was developed to detect circulating antibodies against bovine Theileria in northern China. This is the first report on the seroprevalence of bovine theileriosis in northern China, and it also provides seroepidemiological data on bovine theileriosis in China.

Although only recently recognised, Theileria orientalis (Ikeda) is now the most important infectious cause of anaemia in New Zealand cattle. The aim of this study was to test if vertical transmission of T. orientalis (Ikeda) from dam to calf across the placenta occurs in naturally infected New Zealand dairy cattle and to also test whether the infection status of the dam at calving affects the future susceptibility of its offspring to T. orientalis (Ikeda) infection. Dairy cows (n=97) and their calves were sampled at calving; and the calves again at 4 months of age. All samples were measured for haematocrit and screened for T. orientalis genotypes using a multiplex Buffeli, Chitose and Ikeda specific TaqMan assay. Ikeda positive samples were further tested by singleplex PCR in triplicate to calculate the Ikeda infection intensity as genomes/μl of blood from each infected animal. No T. orientalis (Ikeda) infected calves were born to either T. orientalis (Ikeda) infected or uninfected dams. There were 56/97 dams positive for T. orientalis (Ikeda) infection at calving and 79/90 calves positive for T. orientalis (Ikeda) infection at 4 months of age but no effect on calf susceptibility of dam infection status at calving. There was a significant negative effect of infection intensity on haematocrit after controlling for whether the infected animal was a dam or a 4 month old calf. Vertical trans-uterine transmission of T. orientalis (Ikeda) infection is unlikely in chronically infected dairy cows and thus not a factor in the epidemiology of T. orientalis (Ikeda) infection., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Bovine theileriosis is a serious threat to livestock worldwide. Uncertainty around species prevalence, antigenic diversity and genotypes of strains make it difficult to assess the impact of this parasite and to provide necessary treatment. We aimed to characterize genotypic diversity, phylogeny and prevalence of Theileria orientalis parasites from the states of Telangana and Andhra Pradesh, India by collecting bovine blood samples from the major districts of the two states. Bioinformatic analysis identified antigenic diversity among the prevalent parasite strains using major piroplasm surface protein (MPSP) gene. Our study revealed a prevalence rate of 4.8% (n=41/862) of T. orientalis parasites in bovine animals and a new genotype of T. orientalis parasite which was not previously reported in India. The emergence of these new genotypes could be an explanation for the frequent outbreaks of bovine theileriosis. Further, whole genome sequencing of T. orientalis strains will help to elucidate the genetic factors relevant for transmissibility and virulence as well as vaccine and new drug development., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Theileriosis or East Coast Fever (ECF) is an important livestock disease widespread in Zambia except for some provinces such as Luapula. This freedom status has been achieved due to strict livestock movement regulations that only authorise cattle imports from commercial farms implementing strict ECF control regimens. Recent increases in both the demand and price of beef in Zambia are stimulating a policy change towards a more inclusive inter-provincial trade in live cattle. This may also encourage the introduction of breeding cattle from high production pastoral sectors such as Central Province to stimulate the beef industry in disease free low production areas such as the Luapula Province. To estimate and compare the risks linked with those potential introductions of cattle from the traditional or commercial production sectors of the Central Province, a quantitative risk assessment model was developed. This risk comparison was necessary because the traditional livestock production sector accounts for over 79% of breeding cattle trade in Central Province but is characterised by minimalistic tick-borne disease control and a higher prevalence of ECF. We estimate that should the importation of breeding cattle from Central into Luapula Province be permitted, we could expect to import ECF by the introduction of infected animals at a median rate (5th and 95th percentiles) of every 0.44 years (0.12, 2.60), from the traditional sector compared to every 3.57 years (0.37, 103.6) from the commercial sector. Infected ticks would be expected to enter every 3.46 (0.66, 43.8) years via traditional cattle imports. These risks are strongly influenced by the prevalence of infection, performance of pre-transport screening tests, and the effectiveness of pre-transport tick cleansing. This assessment is expected to provide a model for tick borne disease risk assessments in similar settings, as well as inform ECF control, cattle trade, and stock movement policies in Zambia., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: To determine the distribution of theilerial types in eastern Australian cattle herds and their changing prevalence in regions of New South Wales (NSW)., Design: Survey testing of herds in 2010-11 in Queensland (QLD), NSW and Victoria (VIC) where clinical theileriosis was not evident and ongoing surveillance in NSW through laboratory submissions., Methods: Blood samples were tested by PCR targeting the Theileria orientalis p32 gene and positive tests were examined for the Ikeda, Chitose and Buffeli types. Survey samples from 516 cattle in 50 herds and diagnostic submissions from 434 suspect field cases in 116 herds were analysed., Results: In clinically normal survey cattle, T. orientalis prevalence was high (NSW 23.7%, QLD 56.8%, VIC 34.0%), with variability among regions of each state. Chitose was the most common and widespread type (19.1-43.7% per state), with Buffeli present in all states at a lower prevalence (10.8-24.8% per state). Ikeda was detected in three of five regions in QLD (North, South and South East; prevalence 3.4-15.4%), only one of the surveyed regions in NSW (North Coast; prevalence 74.2%) and in only one animal in VIC. Evidence of clinical disease and laboratory confirmation of Ikeda infection in diagnostic submissions were predominant in several NSW regions, with increasing numbers of affected herds particularly in the coastal Mid-Coast and Cumberland areas., Conclusions: In those regions where prior evidence of theileriosis was uncommon, Ikeda infection was evident in a limited number of NSW regions and multiple regions in QLD. However, clinical disease has continued to become widespread in NSW and VIC, involving Ikeda strains in many regions., (© 2013 Australian Veterinary Association.)

Theileriosis is an important disease in exotic and cross bred dairy cattle in India. Group I was treated with therapeutic drug injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Yakrifit liquid @ 50 mL twice daily for 7 days. Group II was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Yakrifit boli @ 2 boli twice daily for 7 days, Group III was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + AV/SYL/12 @ 50 mL twice daily for 7 days, Group IV was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + AV/SYB/12 @ 2 boli twice daily for 7 days and Group V was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Liv 52 bolus @ 2 boli twice daily for 7 days. In present study, all theileriosis infected groups showed 100 % recovery after treatment with standard drug buparvaquone and adjunct therapy (Yakrifit liquid, Yakrifit boli, AV/SYL/12 liquid, AV/SYL/12 boli and Liv 52 bolus) within 7 days. Within a week, all six animals showed clinical recovery as evaluated on the basis of clinical parameters which returned to normal and improvement in the hematological and biochemical parameters. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a fatal leukemic-like disease of cattle caused by the tick-transmitted protozoan parasite Theileria annulata. The economics of cattle meat and milk production is severely affected by theileriosis in endemic areas. The hydroxynaphtoquinone buparvaquone (BPQ) is the only available drug currently used to treat clinical theileriosis, whilst BPQ resistance is emerging and spreading in endemic areas. Here, we chronically exposed T. annulata-transformed macrophages in vitro to BPQ and monitored the emergence of drug-resistant parasites. Surviving parasites revealed a significant increase in BPQ IC50 compared to the wild type parasites. Drug resistant parasites from two independent cloned lines had an identical single mutation, M128I, in the gene coding for T. annulata cytochrome B (Tacytb). This in vitro generated mutation has not been reported in resistant field isolates previously, but is reminiscent of the methionine to isoleucine mutation in atovaquone-resistant Plasmodium and Babesia. The M128I mutation did not appear to exert any deleterious effect on parasite fitness (proliferation and differentiation to merozoites). To gain insight into whether drug-resistance could have resulted from altered drug binding to TaCytB we generated in silico a 3D-model of wild type TaCytB and docked BPQ to the predicted 3D-structure. Potential binding sites cluster in four areas of the protein structure including the Q01 site. The bound drug in the Q01 site is expected to pack against an alpha helix, which included M128, suggesting that the change in amino acid in this position may alter drug-binding. The in vitro generated BPQ resistant T. annulata is a useful tool to determine the contribution of the various predicted docking sites to BPQ resistance and will also allow testing novel drugs against theileriosis for their potential to overcome BPQ resistance. [ABSTRACT FROM AUTHOR]

Tropical or Mediterranean theileriosis in dairy cattle is widely distributed in many tropical regions of the world. The purpose of this study was to evaluate the proliferation status of mononuclear cells infected with Theileria annulata schizonts in different tissues and its relationship with the pathogenesis of the parasite in cattle by histopathology, immuno-histochemistry and polymerase chain reaction (PCR). Blood and tissue samples of eight Holstein cattle that had been lost due to theileriosis and eight healthy slaughtered cattle of the same breed were collected as a control group after necropsy. The piroplasms in the blood smears and the schizonts in the cytoplasm of the lymphocytes and macrophages of the lymph nodes were microscopically detected. Histopathologically, the proliferation of macrophages, lymphocytes, and plasma cells in lymph nodes and the heart, congestion, and bleeding in the red pulp of the spleen, portal tracts of the liver, interstitial tissue of the kidneys, multifocal necrosis and ulceration in the abomasum together with hyperemia and hemorrhages and lymphoblastic infiltration in the submucosa and lamina propria adjacent to these lesions and emphysema with ecchymotic hemorrhage in the lungs were evident. Immunohistochemistry identified the proliferated cells as mostly Cluster of Differentiation 3- Positive T lymphocytes and macrophage marker antibody 387- positive macrophages. Positive results of PCR for the Tamsl 30.00 kDa gene were observed in lymph nodes, liver, lung and abomasum. It was concluded that the pathological changes were the result of schizont-infected macrophage proliferation leading to severe uncontrolled proliferation of uninfected T lymphocytes. [ABSTRACT FROM AUTHOR]

Simple Summary: Theileriosis is a potentially fatal disease for susceptible cattle and is an important cause of economic losses for farms around the world. Nevertheless, there are animals that can be protected with vaccination or acaricides, or that can recover after treatment is carried out. However, the current trend is to try to reduce the use of drugs and to implement more sustainable strategies. The following review presents the parasite's life cycle, the clinical signs and lesions resulting from infection by Theileria spp. and discusses current strategies to control the disease and the development of strategies based on the genetic selection of resistant animals. Diseases caused by ticks have a high impact on the health, welfare, and productivity of livestock species. They are also an important cause of economic losses in farms worldwide. An example of such diseases is theileriosis, which can be controlled by drugs or vaccines, although these are not fully efficient. Therefore, there is a need to develop alternative and more sustainable and efficient complementary strategies. These may involve the identification and selection of animals more resistant to the disease. Several previous studies have identified significant differences in resistance between different breeds, with resistant breeds typically identified as those native to the region where they are being studied, and susceptible as those from exotic breeds. These studies have indicated that resistance traits are intrinsically related to the modulation of the immune response to infection. This review aims to systematize the general knowledge about theileriosis, emphasize resistance to this disease as a sustainable control strategy, and identify which traits of resistance to the disease are already known in cattle. [ABSTRACT FROM AUTHOR]

Rustaqi cattle breed that living in Al-Qadisiya, and Al-Najaf provinces for decades characterized with their adaptive to harsh environment and diseases. Despite that, there is no clear data about it at Al-Najaf and Al-Qadisiya to resist or not to tick parasites with link to Theileria as it endemic in that region. Therefore, we have designed this study. The current study conducted from October to December, 2020, where 130 blood samples were collected from the Rustaqi native cattle breed, divided into 59 from Al-Qadisiya province and 71 from Al-Najaf regions, as well as we have collected ticks from these animals. Furthermore, in order to understand the immunological association between the presence of the parasite and native cattle that are more resistant to tick-borne diseases, we extracted DNA from blood samples and then used conventional PCR, Sanger sequencing technology. However, the results of the microscopic examination exposed the presence of the parasite inside red blood cells in the form of ring or rod of blood smear taken from the ear vein of cows and the incidence of Theileria in Al-Najaf province was 23.9% and in Al-Qadisiya region was 23.7%, also we have discovered the type of ticks is Hyalomma spp, with statistically significant differences at a significant level P £ 0.05 for the infection close between both provinces. Single nucleotide polymorphisms (SNPs) outcomes uncovered the presence of genetic variants associated with PRKG1 gene that has a role in tick resistance and immune regulation in the original local strains with an inverse correlation between the values of genetic diversity and the difference in nucleotide sequences, which caused a decrease in the proportion of Theileria in the blood. [ABSTRACT FROM AUTHOR]

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]

BACKGROUND: Theileriosis is an important disease of cattle in Iran that could cause economic loss by reducing production and death. The disease in endemic areas would be mild or subclinical in indigenous cattle and clinically acute in exotics OBJECTIVES: This study was conducted to evaluate the resistance of indigenous cattle in comparison with Holstein and crossbred cattle against Theileria annulata through assessment of clinical signs and response to treatment. METHODS: For this purpose, the data history and clinical examination were observed on 51 naturally affected cattle with T. annulata (17 Holstein, 17 crossbred and 17 indigenous cows). Examination of blood smears was used for diagnosis. Detection of schizonts in lymph node punctured smears confirmed validity of the suspect samples. All three groups of cows were treated with buparvaquone along with oxytetracycline. Their clinical signs and the results of blood smear examinations were recorded before and 48 hours after treatment. The severity of clinical signs, parasitemia rates and response to treatment were compared among the groups. RESULTS: The results showed significant difference (P<0.05) in severity of fever, intense and abnormal lung sounds, mucosal changes (pallor, jaundice, petechiae) and ruminal hypomotility among the groups. There was no significant difference in parasitemia rate and response to treatment among the groups. CONCLUSIONS: This study indicated that Iranian indigenous cattle in comparison with Holsteins and crossbreeds had milder clinical manifestations and no significant difference in response to treatment for tropical theileriosis. [ABSTRACT FROM AUTHOR]

Theileria orientalis Ikeda genotype is a parasite that causes a disease in cattle that results in major economic issues in Asia, New Zealand, and Australia. The parasite is transmitted by Haemaphysalis longicornis ticks, which have recently been reported in numerous states throughout the eastern United States. Concurrently, cattle in Virginia showed clinical signs consistent with a hemoprotozoan infection. We used amplicons specific for the major piroplasm surface protein and small subunit rDNA of piroplasms to test blood samples from the cattle by PCR. Bidirectional Sanger sequencing showed sequences with 100% identity with T. orientalis Ikeda genotype 2 sequences. We detected the parasite in 3 unrelated herds and from various animals sampled at 2 time points. Although other benign T. orientalis genotypes are endemic to the United States, detection of T. orientalis Ikeda genotype might represent a risk for the cattle industry in Virginia. [ABSTRACT FROM AUTHOR]

The apicomplexan parasite Theileria orientalis is a tick-borne intracellular protozoan parasite that is widely distributed throughout China. It causes bovine theileriosis in infected cattle, which results in huge economic losses to the cattle industry. In this study, the infection status of T. orientalis was determined in 260 blood samples from cattle from seven provinces across China. Results of a major piroplasm surface protein (MPSP)-PCR assay revealed that an average of 36.5% (95/260) of cattle was positive for T. orientalis infection. Based on the MPSP gene sequences, phylogenetic analysis revealed that these isolates of T. orientalis comprised of eight MPSP types, 1, 2, 3, 4, 5, 7, N1, and N2. This is the first report of new T. orientalis MPSP genotypes N1 and N2 in cattle in China. [ABSTRACT FROM AUTHOR]

Background: Theileria annulata is an economically important cattle disease in North Africa that occurs in subtropical and tropical areas. Accurate and rapid, molecular diagnosis of tropical theileriosis is an important issue that allows early treatment and, prevents transmission. We developed and validated a Theileria annulata specific LAMP assay targeting the cytochrome b multicopy gene, in order to increase the DNA detection sensitivity. Methods: The methodology was used to evaluate the occurrences of T. annulata in 88 field samples collected in Northern Tunisia during 2013-2014. The specificity and sensitivity of the LAMP assays were compared to conventional cytochrome b PCR and routine microscopy commonly used on naturally infected cattle blood samples. Results: The PCR assay showed a sensitivity of 70% and specificity around 75%. Our LAMP assay showed a suitable sensitivity 78.7% and specificity 87.5%, with, however, positive (98.4%) and negative (29.1%) predictive values. Conclusion: The LAMP assay is a simple and convenient diagnostic tool for tropical theileriosis. Moreover, LAMP does not require experienced staff and specialized equipment for sampling procedures and it is practical outside laboratories and can be used for field diagnosis. [ABSTRACT FROM AUTHOR]

Tick-borne diseases (TBDs) impose a significant constraint to livestock production world widely. In this paper, we presented a case of TBD in a cattle farm in Henan, China. 35 blood samples (7 samples sent by veterinarian, 28 samples gathered by our colleagues) were collected from ill, surviving and asymptomatic cattle and microscopic observation and PCR assays were conducted to characterize the pathogens. Genus Ixodes feeding on these cattle were collected and identified. Theileria annulata -like and Anaplasma marginale -like pathogens were observed in the blood smears stained with Giemsa staining under microscope. Furthermore, 5 out of 7 cattle blood samples were found to be positive for T. annulata by PCR. In the 28 blood specimens, three were positive for T. annulata , while A. marginale DNA was detected in nine blood DNA samples. Besides, 56 ticks feeding on cattle were collected from this farm and were all identified as Rhipisephalus microplus , meanwhile, 10 of them were found to be positive for A. marginale . In addition, phylogenetic analysis of the msp4 gene sequences of A. marginale obtained in this study showed that the isolate from cattle (KX840009) fell in the same clade with that of R . microplus (KX904527), sharing 100% similarity. To the best of our knowledge, this is the first confirmed report of outbreak of theileriosis/anaplasmosis in cattle farms in Henan, China. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate serum albumin, glucose, calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), and phosphorus (P) in cows with naturally infected with Theileria annulata. The material of this study was a total of 15 cattle with different ages, breeds and genders diagnosed as theileriosis according to clinical and microscopical examination. According to statistical analyses of biochemical parameters; serum glucose, Ca, Co, Cu, Fe, Mg, and P concentrations were detected significantly low in cattle with theileriosis (P<0.001). In conclusion; mineral substance levels were altered in cattle naturally infected with Theileria annulata. Additionally to the classical treatment of theileriosis, administering mineral supplements including Ca, Co, Fe, Cu, Mg, and P to the animals is thought to be more useful in treatment. [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 hematobiochemical 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<0.01) in hemoglobin, total erythrocyte count, packed cell volume and total leukocyte count, serum glucose, total protein, globulin and albumin level and highly significant increase in aspartate aminotransferase and alanine aminotransferase level as compared to healthy control animals in Group I and II. The animals of Group II treated with buparvaquone along with single blood transfusion shows better recovery then animals of Group I treated with bupavaqone and hematinic. Conclusions: Significant changes were found in hemato-biochemical parameters in theileria affected calves before treatment as compare to healthy control calves. Significant improvement was observed in hemato-biochemical parameters in buparvaquone and single blood transfusion treated calves as compare to another group, so it is concluded that buparvaquone and single blood transfusion is better combination for treatment of theileriosis. [ABSTRACT FROM AUTHOR]

Theileriosis caused by Theileria lestoquardi (malignant ovine theileriosis) in sheep and Theileria annulata (tropical theileriosis) in cattle is an important hemoprotozoal tick-borne disease in Iran. Due to major biologic and phylogenic similarities of these two species, this study was carried out to investigate the occurrence of natural infections with T.lestoquardi and T.annulata in cattle with clinical theileriosis in Ahvaz, southwest Iran. Fifty one cattle were selected based on clinical signs of theileriosis and confirmation by microscopic examination of blood smears. Blood samples were collected from each animal and hematologic and microscopic examinations were performed. Theileria piroplasmic forms were detected in all affected cattle. Pale mucous membranes (43.14%), icterus (11.76%) and fever (70.6%) were also observed. PCR-RFLP analysis revealed T. annulata infection in all tested cattle while coinfections with T. lestoquardi were found in two samples (3.92%). All sampled cattle including the two with mixed species Theileria infection were anemic. This is the first report of Theileria species cross infections in cattle with clinical theileriosis in Iran. It can be concluded that cattle can be infected with both pathogenic Theileria species, T. lestoquardi and T. annulata which can be an important issue in the epidemiology and spread of ovine malignant theileriosis. [ABSTRACT FROM AUTHOR]

This study was aimed at determining the prevalence of T. annulata and T. buffeli/orientalis species in cattle from Kirikkale province using reverse line blotting (RLB) and comparing the results with microscopy. Blood samples were collected from 294 cattle from 9 districts of Kirikkale (Centre, Bahsili, Baliseyh, Celebi, Delice, Karakecili, Keskin, Sulakyurt and Yahsihan) between May-October 2010. From cattle that has at least grazed once and over one year old of age. A blood smear from ear tip peripheral blood was prepared and a venous blood sample from jugular vein was taken. Blood smears were inspected and documented for the presence of Theileria spp. piroplasms after Giemsa staining. Venous blood samples were used for extraction of DNA for Reverse Line Blotting (RLB) analyses. Out of 294 samples analyzed 44 (15%) were found to have Theileria spp. piroplasms with microscopic examination, while 77 (26.1%) harbored T. annulata as determined by RLB. In none of the samples T. buffeli/orientalis was detected. The prevalence of Theileria spp. and T. annulata was highest in Keskin district. This study presents the first investigation of Theileria species in cattle from Kirikkale province. [ABSTRACT FROM AUTHOR]

Babesiosis, theileriosis, and anaplasmosis are the tick-borne diseases of cattle in most of the tropical areas and Turkey. A total of 270 cattle were randomly selected from 27 villages and 77 farms in the Black Sea Region of Turkey for the determination of infections by Babesia bigemina and Anaplasma marginale by using the indirect fluorescent antibody (IFA) test and the competitive enzyme-linked immunosorbent assay (cELISA). Of those, 242 and 256 sera were also analyzed for antibodies against Theileria annulata and Babesia bovis infections using the IFA test, respectively, during the period of June 2006 to June 2007. Of the 256 tested sera, 99 (38.6%) were positive for B. bovis; from the total 270 sera, 40 (14.8%) were positive for B. bigemina; of the 242 sera tested, 31 (12.8%) were positive for T.annulata by IFA; and from the total 270 sera, 102 (37.8%) were positive for A. marginale by cELISA. A total of 1125 ixodid ticks were collected and 10 species were identified. Antibodies produced by infections with Babesia spp., Anaplasma spp., and Theileria spp. were found highly prevalent. Infections caused by the agents and the tick species may cause severe economic damage to cattle production from the Black Sea Region of Turkey. [ABSTRACT FROM AUTHOR]

Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (ß-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata. [ABSTRACT FROM AUTHOR]

AIMS: To describe the epidemiology of the epidemic of bovine anaemia associated withTheileria orientalisinfection (TABA) in New Zealand between 30 August 2012 and 4 March 2014. METHODS: Blood samples and associated data were obtained from cases of TABA. The case definition for TABA was met when piroplasms were present on blood smears and the haematocrit was ≤0.24 L/L. Samples were analysed using quantitative PCR (qPCR) assays for the detection ofT. orientalisIkeda type. Only cases that were positive in the qPCR assays were included in the analysis. A case herd was defined as a herd that had ≥1 animal positive forT. orientalisIkeda. Movement records for farms were accessed through the national animal identification and tracing scheme. The OR for cattle movements onto a case farm compared to a non-case farm was estimated using a generalised estimating equation model and the geodesic distance for movements onto case and non-case farms compared using Student'st-test. The kernel-smoothed risk of disease at the farm level was calculated using an extraction map and the clustering of diseased farms in time and space was measured using the spatial temporal inhomogeneous pair correlation function. RESULTS: In the first 18 months there were 496 case herds; 392 (79%) were dairy and 104 (21%) beef herds. Of 882 individual cases, 820 (93.0%) were positive forT. orientalisIkeda in the qPCR assays. Case herds were initially clustered in the Northland, then the Waikato regions. The OR for a case farm compared to a non-case farm having ≥1 inward cattle movements was 2.03 (95% CI=1.52–2.71) and the distance moved was 26 (95% CI=20.8–31.3) km greater for case farms. The risk of disease was highest in a north, north-eastern to south, south-western belt across the Waikato region. The spatial-temporal analysis showed significant clustering of infected herds within 20–30 days and up to 15 km distant from a case farm. CONCLUSIONS:Theileria orientalisIkeda type is likely to have been introduced into regions populated with naïve cattle by the movement of parasitaemic cattle from affected areas. Local spread through dispersed ticks then probably became more important for disease transmission between herds once the disease established in a new area. CLINICAL RELEVANCE: Dairy and beef farming in the North Island of New Zealand will be significantly changed in the coming years by the incursion of this new disease. [ABSTRACT FROM AUTHOR]

AIM: To estimate incidence risk, cumulative mortality and case fatality rate within herds affected by bovine anaemia associated withTheileria orientalisinfection (TABA), in New Zealand during the early phase of the epidemic (August 2012–September 2013). METHODS: A mixed methods approach was utilised to integrate data from various sources, including a detailed questionnaire carried out on 18 dairy farms which had experienced cases of TABA; a brief telephone survey of an additional 139 case farms; and data extracted from a Ministry for Primary Industries database for a further 42 case farms. The subsequent analysis determined incidence risk, cumulative mortality and case fatality rates for beef and dairy herds. RESULTS: Data were analysed from 196/263 (74%) known case farms at the date of closing the questionnaires. These farms contained 99,505 cattle; 2,847 animals were reported with clinical disease, and a further 590 animals were recorded as having died from TABA. The within-herd incidence risk, cumulative mortality and case fatality rate were consistent between the three data sources, did not differ between beef and dairy herds, and were estimated to be 0.97 (inter-quartile range (IQR) 0.36–2.07)%, 0.23 (IQR 0.00–0.66)% and 16.67 (IQR 0.00–33.33)%, respectively. There was substantial variability in the level of impact, with 22 farms severely affected (incidence risk >5% and cumulative mortality >5%). CONCLUSIONS: The mixed methods approach was effective in dealing with the disparate data sources. The inclusion of the majority of farms known to be affected at the time the questionnaires were performed implies that the information is likely to be representative. The collective outputs of the analyses represent the best estimate available of within-herd measures of disease frequency in the early phase of the epidemic in New Zealand. The limitations of the data imply that their primary application may be to inform the design of subsequent structured observational field studies. CLINICAL RELEVANCE: The results of this study provide information on the impact of TABA on cattle farms during the emergence and early spread of the disease, as well as for generating hypotheses on causal mechanisms and risk factors that may influence the course of disease. [ABSTRACT FROM AUTHOR]

AIM: To describe the prevalence and spatial distribution of cattle herds infected with Ikeda and non-Ikeda types ofTheileria orientalisin New Zealand between November 2012 and June 2013. METHODS: Pooled serum samples collected historically between November 2012 and June 2013 were obtained from cattle herds throughout New Zealand. Each pooled sample consisted of approximately 20 individual cattle samples from that herd, and was provided with details of the spatial location of the herd (n=722). DNA from all samples was tested using two quantitative PCR assays for the detection ofT. orientalis(all types) and the Ikeda type. The proportion of herds that were positive forT. orientalisand Ikeda type, or that were positive forT. orientalisbut negative for Ikeda type (non-Ikeda positive) was determined for different regions of New Zealand. RESULTS: The highest prevalence of herds infected with Ikeda type was detected in the Northland (33/35; 94%) and Auckland and the Waikato (63/191; 33%) regions. Only 2/204 (1%) herds were positive for the Ikeda type in the South Island. A high percentage of herds that were positive for non-Ikeda types was detected in the Gisborne and Hawkes Bay (23 (95%CI=13–37)%), Auckland and Waikato (22 (95%CI=16–29)%) and Bay of Plenty (24 (95%CI=10–44)%) regions. CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of Ikeda type detected in cattle herds in the Northland, Auckland and Waikato regions represents a risk to naive cattle being introduced into these regions. There is also the potential for resident cattle herds in the Gisborne and Hawkes Bay, Auckland, Waikato and Bay of Plenty regions to experience increased infection with the Ikeda type. The overall impact experienced by regions will depend on other factors such as the number of herds present and the predominant type of farming, as well as the interplay between tick ecology, cattle immunity and movement patterns of cattle. [ABSTRACT FROM AUTHOR]

Aim: In India, dairy industries are important for the livelihood of small scale farmers and dairy owners. Tropical theileriosis, mostly affecting dairy cattle and buffaloes is a major threat to dairy and related industries. Tropical theileriosis is caused by Theileria annulata, a hemoprotozoan parasite transmitted by Ixodid ticks of Hyalomma spp. In the present study, we examined the clinical signs, hematological parameters and flow cytometric profile of whole blood in 30 theileriosis affected crossbred cattle. The aim of our study is to analyze, in comparison with clinical and hematological diagnosis, whether flow cytometry based profiling of leukocytes could be used as better, quick and alternative method for diagnosis and screening of bovine tropical theileriosis in crossbred cattle. Materials and Methods: In this study, we screened parasites in 30 peripheral blood samples from clinical cases of theileriosis by Giemsa's staining technique in crossbred cattle. Hematological analysis was done to estimate hemoglobin (Hb) content, total red blood cell (RBC) count, total leukocyte count and differential leukocyte count. Further, flow cytometric analysis of whole blood was carried out to study leukocytes profile in affected cattle. Results: Microscopic examination of stained blood films revealed the presence of piroplasms in erythrocytes and schizonts in lymphocytes. Hematological examination revealed significant (p<0.05) decrease of Hb percent (Hb %), reduced total RBC and total leukocytes, lymphocytosis, eosinopenia, and neutropenia compared to that of apparently healthy cattle. Flow cytometric profiling of leukocytes revealed the severe effect on shape, size, and granularity of leukocytes, marked decrease in granulocytes and 3-5 fold increase in lymphocytes count compared to clinically healthy cattle. Thus, in both methods, namely conventional and flow cytometric analysis, marked lymphocytosis and decrease in other blood cell counts were observed compared to clinically healthy cattle. Conclusions: From results, it can be concluded that though conventional staining techniques and hematology are efficient in diagnosis of theileriosis, leukocytes profiling based on flow cytometry combined with clinical examination could be a quick, novel and alternative method for diagnosis and screening of clinical tropical theileriosis in crossbred cattle. Thus, there is potential to offer a flow cytometry based diagnostic service for tropical theileriosis in crossbred cattle. [ABSTRACT FROM AUTHOR]

Aim: The aim of the present study was to investigate the presence of Theileria in blood samples of crossbred and indigenous adult cows raised under unorganized small scale farming system in a Babesia and Anaplasma endemic geographical area from Assam, India and to see its transmission through Rhipicephalus (Boophilus) microplus ticks. Materials and Methods: For the present study, 57 clinical cases of cattle suspected to be of hemoparasitic infections were taken into consideration. The parasites were identified based on morphology in giemsa stained blood smear followed by polymerase chain reaction (PCR). Sera samples were tested for T. annulata antibodies in plate and Dot-ELISA. PCR was also conducted in eggs of Rhipicephalus (Boophilus) microplus tick collected from a Theileria orientalis positive animal. Results: PCR amplified 1124, 776, and 160 bp DNA fragments of B. bigemina (64.91%), T. orientalis(21.05%) and A. marginale (14.03%), respectively. This assay further conducted in 12 T. orientalis positive blood samples with primers of Buffeli, Chitose, and Ikeda variants of T. orientalis showed 3 samples positive to Ikeda type and none for Buffeli and Chitose. Babesia bovis and Theileria annulata specific primers also did not amplify any fragment during the PCR assay of the blood samples. Further, all sera samples tested negative to T. annulata antibodies in Plate and Dot-ELISA. PCR conducted in eggs of R (B).microplus tick collected from a T. orientalis positive animal revealed presence of the parasite DNA. Gradual improvement in physical condition leading to complete recovery in 10 out of 12 T. orientalis infected clinical cases treated with buparvaquone(at 2.5mg/kg.b.wt I/M) was the feedback obtained from field veterinarians and the cattle owners. Conclusion: The present investigation represents the first report of occurrence of T. orientalis in cattle of Assam with involvement of pathogenic Ikeda strain in clinical outbreaks and its possible natural transmission by R (B). microplus through the transovarian mode. [ABSTRACT FROM AUTHOR]

Background: Theileria are blood-borne intracellular protozoal parasites belonging to the phylum Apicomplexa. Previously considered a benign parasite in Australia, outbreaks of clinical disease resulting from Theileria orientalis genotypes have been reported in Australia since 2006. Since this time, outbreaks have become widespread in south-eastern Australia, resulting in significant adverse impacts on local dairy and beef industries. This paper provides the first investigation into the possible biological and mechanical vectors involved in the rapid spread of the parasite. Methods: To identify possible vectors for disease, ticks, biting flies and mosquitoes were collected within active outbreak regions of Gippsland, Victoria. Ticks were collected from cattle and wildlife, and mosquitoes and biting flies were collected in traps in close proximity to outbreak herds. Ticks were identified via DNA barcoding of the mitochondrial cytochrome oxidase I gene. Barcoded ticks were pooled according to species or phylogenetic group and tested for the presence of T. orientalis and the genotypes Ikeda, Chitose and Buffeli using real-time PCR. Results: DNA barcoding and phylogenetic analysis identified ticks from the following species: Haemaphysalis longicornis, Ixodes holocyclus, Ixodes cornuatus, Ixodes hirsti, and Bothriocroton concolor. Additional Haemaphysalis, Ixodes and Bothriocroton spp. were also identified. Of the ticks investigated, only H. longicornis ticks from cattle carried theilerial DNA, with the genotypes Ikeda, Chitose and Buffeli represented. Mosquitoes collected in close proximity to outbreak herds included; Aedes camptorhynchus, Aedes notoscriptus, Coquillettidia linealis, Culex australicus, and Culex molestus. Low levels of T. orientalis Buffeli genotype were detected in some mosquitoes. The haematophagous flies tested negative. Conclusions: This is the first demonstration of a potential vector for T. orientalis in the current Australasian disease outbreak. [ABSTRACT FROM AUTHOR]

Background: Oriental theileriosis is a tick-borne disease of bovines caused by the members of the Theileria orientalis complex. Recently, we developed a multiplexed tandem (MT) PCR to detect, differentiate and quantitate four genotypes (i.e., buffeli, chitose, ikeda and type 5) of T. orientalis. In this study, we used MT PCR to assess the prevalence and infection intensity of four T. orientalis genotypes in selected cattle herds that experienced oriental theileriosis outbreaks in New Zealand, and compared the sensitivities and specificities of MT PCR, PCR-high resolution melting (PCR-HRM) and a TaqMan® qPCR. Methods: MT PCR, PCR-HRM analysis for T. orientalis and a TaqMan® qPCR assay for ikeda genotype were employed to test 154 and 88 cattle blood samples from North (where oriental theileriosis outbreaks had occurred; designated as Group 1) and South (where no outbreaks had been reported; Group 2) Islands of New Zealand, respectively. Quantitative data from MT PCR assay were analyzed using generalized linear model and paired-sample t-test. The diagnostic specificity and sensitivity of the assays were estimated using a Bayesian latent class modeling approach. Results: In Group 1, 99.4% (153/154) of cattle were test-positive for T. orientalis in both the MT PCR and PCR-HRM assays. The apparent prevalences of genotype ikeda in Group 1 were 87.6% (134/153) and 87.7% (135/154) using the MT PCR and Ikeda TaqManW qPCR assays, respectively. Using the MT PCR test, all four genotypes of T. orientalis were detected. The infection intensity estimated for genotype ikeda was significantly higher (P = 0.009) in severely anaemic cattle than in those without anaemia, and this intensity was significantly higher than that of buffeli (P <0.001) in the former cattle. Bayesian latent class analysis showed that the diagnostic sensitivities (97.1-98.9%) and specificities (96.5-98.9%) of the three PCR assays were very comparable. Conclusion: The present findings show the advantages of using the MT PCR assay as a useful tool for in-depth epidemiological and transmission studies of T. orientalis worldwide. [ABSTRACT FROM AUTHOR]

Live attenuated vaccines are used to combat tropical theileriosis in North Africa, the Middle East, India, and China. The attenuation process is empirical and occurs only after many months, sometimes years, of in vitro culture of virulent clinical isolates. During this extensive culturing, attenuated lines lose their vaccine potential. To circumvent this we engineered the rapid ablation of the host cell transcription factor c-Jun, and within only 3 weeks the line engineered for loss of c-Jun activation displayed in vitro correlates of attenuation such as loss of adhesion, reduced MMP9 gelatinase activity, and diminished capacity to traverse Matrigel. Specific ablation of a single infected host cell virulence trait (c-Jun) induced a complete failure of Theileria annulata–transformed macrophages to disseminate, whereas virulent macrophages disseminated to the kidneys, spleen, and lungs of Rag2/γC mice. Thus, in this heterologous mouse model loss of c-Jun expression led to ablation of dissemination of T. annulata–infected and transformed macrophages. The generation of Theileria-infected macrophages genetically engineered for ablation of a specific host cell virulence trait now makes possible experimental vaccination of calves to address how loss of macrophage dissemination impacts the disease pathology of tropical theileriosis. Author Summary: Tropical theileriosis is a leukaemia-like disease of cattle caused by the Apicomplexa parasite Theileria annulata. Live attenuated vaccines are used to control the mortality and morbidity of tropical theileriosis and each endemic country produces its own vaccine by isolating a virulent clinical isolate and then growing the infected and transformed macrophages for many months in the laboratory. With time, virulence is progressively lost, and loss is evaluated from time to time by injecting infected macrophages into calves and monitoring any resulting pathology. If the calves demonstrate clinical symptoms, in vitro culture of vaccine lines continues, and eventually attenuation is achieved after approximately two years. The process is empirical, and with time in culture the infected macrophage lines lose their vaccine potential such that when cattle are injected the immune response to the vaccine can be compromised. To circumvent these problems we engineered the rapid loss of a host macrophage virulence trait and obtained complete attenuation of dissemination of T. annulata–transformed macrophages when tested in an immune-compromised mouse model. This rational, rather than empirical, approach now allows injecting into calves Theileria-infected macrophages engineered for attenuation to evaluate the role of Theileria-infected macrophage dissemination in the pathology of tropical theileriosis. [ABSTRACT FROM AUTHOR]

Background Oriental theileriosis is a tick-borne, protozoan disease of cattle caused by members of the Theileria orientalis-complex. Recent outbreaks of this disease in eastern Australia have caused major concerns to the dairy and beef farming communities, but there are no published studies of the economic impact of this disease. On a farm in Victoria, Australia, we assessed whether oriental theileriosis has an impact on milk production and reproductive performance in dairy cows. Methods Blood samples collected from all 662 cows on the farm were tested using an established molecular test. For individual cows, milk production and reproductive performance data were collected. A clinical assessment of individual cows was performed. Based on clinical findings and molecular test results, the following groups of cows were classified: group 1, with cardinal clinical signs of oriental theileriosis and molecular test-positive for T. orientalis; group 2, with mild signs of theileriosis and test-positive; group 3, with no clinical signs and test-positive; and group 4, with no clinical signs and test-negative. Milk production and reproductive performance data for groups 1, 2 and 3 were each compared with those for group 4 using linear and logistic regression analyses, respectively. Results At 100 days of lactation, group 1 cows produced significantly less milk (288 l; P = 0.001), milk fat (16.8 kg; P < 0.001) and milk protein (12.6 kg; P < 0.001) compared with group 4. At this lactation point, group 2 also produced significantly less milk fat (13.6 kg; P = 0.002) and milk protein (8.6 kg; P = 0.005) than group 4. At 305 days of lactation, group 1 cows produced significantly less milk (624 l; P = 0.004), milk fat (42.9 kg; P < 0.001) and milk protein (26.0 kg; P < 0.001) compared with group 4 cows. Group 2 cows also produced significantly less milk fat (21.2 kg; P = 0.033) at this lactation point. No statistically significant difference in reproductive performance was found upon pairwise comparisons of groups 1-3 with group 4 cows. Conclusions The present findings demonstrate that clinical oriental theileriosis can cause significant milk production losses in dairy cattle. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the relationship between the degrees of anemia and blood gases in cattle with theileriosis. It included 28 cattle with theileriosis and 7 healthy animals as the control group. The cattle with theileriosis were divided into 4 groups according to their hematocrit (Hct) values: cattle with Hct of >26 were considered as nonanemic (Group 1), Hct of 20--26 as mildly anemic (Group 2), Hct of 14--19 as moderately anemic (Group 3), and Hct of 10--13 as severely anemic (Group 4). The PCO2, HCO3, and TCO2 levels in the study groups were lower than those of the control group. Moreover, reductions in HCO3 and TCO2 concentrations were significant only in the severely anemic group. Similarly, the lowest PCO2 levels were also detected in the severely anemic group. There were no significant differences in the SO2 levels between the control and study groups. On the other hand, the increase was significant in the severely anemic group when compared to Groups 1 and 2. The results seemed to indicate the trend of uncompensated metabolic acidosis in theileriosis cases with severe anemia; thus, veterinary surgeons should bear this in mind when evaluating the prognosis for such cases. [ABSTRACT FROM AUTHOR]