This study aims to evaluate the responses of peripheral blood monocyte-derived macrophages (PBMDMs) from Theileria orientalis carrier cattle following exposure to Pasteruella multocida B:2 (PM B:2) and latex beads. Twenty-six male crossbred Kedah-Kelantan (KK) cattle were sampled for this study and quantitative PCR (qPCR) was employed in the detection of T. orientalis MPSP gene. Bactericidal assay using a 10:1 multiplicity of infection was performed to measure the phagocytosis and intracellular killing of PM B:2 by PBMDMs. The cell cultures were inoculated with 107 cfu/mL of PM B:2 and incubated in a humidified incubator. The absence of clinical signs, previous history of T. orientalis infection and an MPSP gene copy number below 15,000 GC/μL suggest that the cattle were asymptomatic chronic carriers. A non-significant phagocytic and mean cell death rates were observed in the PBMDMs of T. orientalis positive cattle relative to clinically healthy cattle (CHC) (p > 0.05). The PBMDMs of T. orientalis positive cattle had the lowest mean rate of intracellular killing relative to the CHC at the 30th minute post-infection only (p < 0.05). Exposure to latex beads caused an increase in the appearance of multinucleated macrophages following incubation of PBMDMs from T. orientalis positive cattle. Furthermore, the phagocytic index of PBMDMs of T. orientalis positive cattle were low or poor compared to that of CHC (p = 0.000). Therefore, our findings suggest that PBMDMs from cattle with chronic T. orientalis infection can efficiently phagocytise and kill PM: B2 but exhibited poor phagocytosis ability for foreign bodies despite appearance of multinucleated macrophages.

Theileriae are obligate intracellular protozoan parasites which are transmitted by ixodid ticks and infect both wild and domestic ruminants worldwide. Theileriosis causes significant economic losses to the livestock industry in many countries due to the high morbidity and mortality in cattle herds. In Russia, information concerning prevalence of Theileria spp. in cattle is very limited. This study reports on molecular characterization and phylogenetic analysis of Theileria spp. parasites detected in cattle from the Moscow region of Russia. Phylogenetic analysis based on the full length 18S rRNA gene revealed that the Russian Theileria parasites belong to the Theileria orientalis / Theileria buffeli / Theileria sergenti group and share a common genotype with T. buffeli Marula from Kenya, T. buffeli isolates from Japan and South Korea, T. orientalis isolate from Australia and T. sergenti isolate from Japan, which belong to the pathogenic Chitose genotype.

BACKGROUND: In regions with endemic Theileria orientalis, recovered cattle are carriers, leaving newborn calves and introduced stock as the most susceptible groups to develop clinical disease after tick infestation with parasite transmission.
METHODS: To gain information on the kinetics of infection and development and effects of theileriosis caused by virulent ikeda and chitose genotypes and the \"benign\" buffeli genotype of T.orientalis, this study sampled a total of 134 calves from 3 farms in Dorrigo, Australia, a region with multiple genotypes of the parasite. In addition, 30 introduced beef weaners were bled and weighed for 6 months after arrival.
RESULTS: In both cohorts, parasitaemia with the ikeda and chitose (genotypes 1 & 2) of T.orientalis was detectable by PCR within 3-4 weeks in >95 % of the groups, with maximal gene copies generated around 5-7 weeks after birth or introduction, before declining. In contrast, parasitaemias of T.orientalis buffeli (genotype 3), increased slowly, with around 80 % of the population testing positive by PCR after 4 months. The parasitaemias of the buffeli genotype did not exceed 40,000 gene copies /ul and were similar to those exhibited by ikeda and chitose genotypes following the \"first wave of parasitaemia\" as cattle entered the carrier state. In the 30 introduced weaners, the early infection with T.orientalis ikeda and chitose also caused significant reductions in packed cell volume (PCV) and incurred an estimated 20Kg loss in weight gain; the latter had not been recovered by 6 months after introduction.
CONCLUSIONS: The results support previous findings in other endemic regions of theileriosis, and imply that amelioration of the early high levels of parasitosis by the virulent genotypes could reduce the initial impact of theileriosis on production, which appears much less affected once animals enter the carrier state.

The aims of this study were to monitor the change in Theileria orientalis Ikeda type infection intensity, haematocrit, milk production and reproduction on three New Zealand spring calving dairy herds, over the 2014-2015 milking season. Three spring calving dairy farms, A, B and C, from high risk (endemically stable), low risk (endemically unstable), and zero risk (disease-free) tick areas respectively were followed through the 2014-2015 milking season. On Farms, A and B, 100 cows were randomly selected at the first visit, and the same cows blood sampled every month thereafter, whilst on Farm C, the whole herd was blood sampled bimonthly (140 cows). Blood samples were tested for haematocrit, by centrifugation, and Ikeda infection intensity, using qPCR. Animals that were Ikeda type PCR positive at the first sampling were described as prevalence cases and cows that were negative at the first sampling and became PCR positive during the sampling period were described as incidence cases. Production and reproduction data were accessed through LIC MINDA® and milk production data was standardised to energy corrected milk (ECM). In addition, the effect of buparvaquone (BPQ) treatment on milk production was estimated on Farm B. The prevalence of infection at the first sampling was 100 % on Farm A, 57 % on Farm B and 26 % on Farm C. The incidence risk of infection over the sampling period on Farms B and C was 25 % and 2 % and the incident rate was 0.026 and 0.002 cases per cow-month respectively. The average infection intensity for prevalence cases on all farms was low throughout the milking season, <7000 Ikeda organisms/μL however, cases of anaemia still occurred. There was no direct effect of infection intensity on milk production or from being a prevalence case compared to an uninfected cow on milk production, across all farms. However, on Farm B there was a loss of 266 kg (95 % CI 82 ̶ 450) ECM (∼20 kg milk solids) for incidence cases and a loss of 458 kg (95 % CI 211 ̶ 710) of ECM for buparvaquone treated cows, compared to uninfected cows. No significant effect of Ikeda infection on reproduction could be shown for Farms B and C, reproductive data for Farm A was not available. The effect of T. orientalis Ikeda type infection on production and reproduction appears to be minimal once animals have passed through the acute phase of infection and reached the chronic, asymptomatic carrier phase of infection.

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.