Besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates

Background Bovine besnoitiosis, caused by the protozoan Besnoitia besnoiti, reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of besnoitiosis. Methods We compared invasion and proliferation of one B. tarandi (from Finland) and seven B. besnoiti isolates (Bb-Spain1, Bb-Spain2, Bb-Israel, Bb-Evora03, Bb-Ger1, Bb-France, Bb-Italy2) in MARC-145 cell culture. Host cell invasion was studied at 4, 6, 8 and 24 h post infection (hpi), and proliferation characteristics were compared at 24, 48, 72, 96, 120, and 144 hpi. Results In Besnoitia spp., the key parameters that determine the sequential adhesion-invasion, proliferation and egress steps are clearly distinct from those in the related apicomplexans Toxoplasma gondii and Neospora caninum. Besnoitia spp. host cell invasion is a rather slow process, since only 50 % of parasites were found intracellular after 3–6 h of exposure to host cells, and invasion still took place after 24 h. Invasion efficacy was significantly higher for Bb-France, Bb-Evora03 and Bb-Israel. In addition, the time span for endodyogeny to take place was as long as 18–35 h. Bb-Israel and B. tarandi isolates were most prolific, as determined by the tachyzoite yield at 72 hpi. The total tachyzoite yield could not be predicted neither by invasion-related parameters (velocity and half time invasion) nor by proliferation parameters (lag phase and doubling time (dT)). The lytic cycle of Besnoitia was asynchronous as evidenced by the presence of three different plaque-forming tachyzoite categories (lysis plaques, large and small parasitophorous vacuoles). Conclusions This study provides first insights into the lytic cycle of B. besnoiti isolates and a standardised in vitro model that allows screening of drug candidates for the treatment of besnoitiosis.

Molecular mechanism of jet fuel induced immune suppression

Dermal exposure to jet fuel suppresses the immune response. Immune regulatory cytokines, and biological modifiers, including platelet activating factor, prostaglandin E2, and interleukin-10 have all been implicated in the pathway leading to immunosuppression. It is estimated that approximately 260 different hydrocarbons are found in JP-8 (jet propulsion-8) jet fuel, and the identity of the immunotoxic compound is not known. The recent availability of synthetic jet fuel (S-8), which is devoid of aromatic hydrocarbons, made it feasible to design experiments to test the hypothesis that the aromatic hydrocarbons are responsible for jet fuel induced immune suppression. Applying S-8 to the skin of mice does not up-regulate the expression of epidermal cyclooxygenase-2 nor does it induce immune suppression. Adding back a cocktail of 7 of the most prevalent aromatic hydrocarbons found in jet fuel to S-8 up-regulated cyclooxygenase-2 expression and induced immune suppression. Cyclooxygenase-2 induction can be initiated by reactive oxygen species (ROS). JP-8 treated keratinocytes increased ROS production, S-8 did not. Antioxidant pre-treatment blocked jet fuel induced immune suppression and cyclooxygenase-2 up-regulation. Accumulation of reactive oxygen species induces oxidant stress and affects activity of ROS sensitive transcription factors. JP-8 induced activation of NFκB while S-8 did not. Pre-treatment with antioxidants blocked activation of NFκB and parthenolide, an NFκB inhibitor, blocked jet fuel induced immune suppression and cyclooxygenase-2 expression in skin of treated mice. p65 siRNA transfected keratinocytes demonstrated NFκB is critically involved in jet fuel induced COX-2 expression. These findings clearly implicate the aromatic hydrocarbons found in jet fuel as the agents responsible for inducing immune suppression, in part by the production of reaction oxygen species, NFκB dependent up-regulation of cyclooxygenase-2, and the production of immune regulatory factors and cytokines. ^

Occurrence of late Aptian/earliest Albian radiolaria from ODP Hole 113-693A (Table 2)

Unusually well preserved Cretaceous radiolarians are observed in the subsurface sections from two drilled sites in the Weddell Sea collected during Leg 113 of the Ocean Drilling Program. Radiolarians from the lithified calcareous chalk of Hole 689B represent the first Campanian-Maestrichtian assemblage which is characterized by abundant Cromyodruppa Iconcentrica, Dictyomitra multicostata, and Protostichocapsa stocki. Abundant Pseudodictyomitra pentacolaensis and Diacanthocapsa sp. 1, on the other hand, are the main constituents of the assemblage from the latest Aptian/earliest Albian diatomite of Hole 693B. These represent the oldest and the highest-latitude reported radiolarian occurrences from the Atlantic sector of the Antarctic Ocean. The assemblages are marked by their low diversity and an absence of low- to mid-latitude zonal indices.

Planktonic foraminifera of the Mediterranean Sea

A high-resolution biochronology is presented for the Late Quaternary of the central Mediterranean. In the Late Pleistocene-Holocene successions three assemblage zones are distinguished on the basis of frequency patterns of planktic foraminifera. The age of these zones is determined by Accelerator Mass Spectrometry (AMS)14C dating. The zonal boundaries are dated at 12,700 yr B.P. (the end of Termination Ia) and 9600 yr B.P. (the start of Termination Ib), respectively. The AMS dates show that major changes in the planktic and benthic realms occurred synchronously over wide areas, although records of individual species may show important regional differences. In the studied areas, resedimentation processes revealed by anomalous successions of14C dates, play a far more important role than indicated by the sedimentological and micropaleontological data. Possibly these processes contribute to the very high accumulation rates in the glacial Zone III. Although the AMS technique has increased the accuracy of14C-measurements, admixture of older carbonate may still lead to substantial age differences between areas with different sedimentary regimes.