The invasion process of bovine erythrocyte by Babesia divergens: knowledge from an in vitro assay

ABSTRACT: Babesia divergens is a tick-transmitted apicomplexan parasite for which asexual multiplication in its vertebrate hosts is restricted to erythrocytes. Current knowledge of invasion of these target cells is limited. An efficient in vitro invasion assay was set up to gain access to this information. Parasites prepared from infected RBC, lysed by electroporation, and mixed with bovine RBC in a selected synthetic medium (RPMI 1640 supplemented with calcium) were able to establish subsequent cultures with parasitemia ranging from 6 to 14%. Free parasites remaining in the invasion medium could be eliminated by Percoll gradient and culture could be pursued with the freshly invaded erythrocytes. In this way, the invasion time window could be shortened to obtain a synchronised start of the culture or to study the kinetics of invasion. With this assay we demonstrate that 1) erythrocyte invasion by B. divergens is a rapid process since 70% of the invasion-competent parasites invaded the RBC in less than 45 s; 2) all invasion-competent parasites achieved invasion within 10 min of contact; 3) one erythrocyte could be invaded concomitantly by two merozoites; 4) despite a synchronous start, the parasite population evolved heterogeneously resulting in a progressive loss of synchronisation. Western blot analysis of proteins collected from invasion medium were performed with sera from animals experimentally infected with B. divergens and highlighted several proteins. The dose-dependent, inhibitory effects of these sera on B. divergens invasion suggest that these proteins might be involved in the invasion process. Further investigations are required for their characterisation.     

River bed restoration boosts habitat mosaics and the demography of two rare non-aquatic vertebrates

Many river revitalization projects are currently under development, but we still lack proper assessments of their effects on biodiversity, especially regarding non-aquatic vertebrates. We analysed the progressive changes in habitat mosaics, and the demographic response of two rare species of wading birds along a Swiss section of the Rhone river over a period of three decades. During the period preceding revitalization, the diversity of the habitat mosaic had been steadily declining. This impoverishment could however be reverted after 1994, when the revitalization was initiated: in 2006, the re-established habitat matrix was more diverse than at any time during the previous 27 years. The formerly channelized stream underwent radical transformation as it mutated into a heterogeneous braided river boosting pioneer riverine habitats. This has led the two wading bird species to establish significantly more territories than before the revitalization operations. Population size of Actitis hypoleucos and Charadrius dubius increased by 83% and 20%, respectively, between 1995 and 2007. During the same period, A. hypoleucos declined dramatically in Switzerland as a whole, whilst C. dubius showed no distinct population trend. The response of the two waders may have resulted from an increase in the number of lateral arms of the river, where the availability of productive temporary ponds provides excellent foraging opportunities. A significant proportion of these habitats was created by a targeted extraction of sediments with the objective to promote biodiversity while maintaining economic activity and improving security against floods. This suggests that gravel exploitation is not incompatible with efficient river restoration.     

Development and evaluation of a new method for sampling and monitoring the symphylid population in pineapple

BACKGROUND: Symphylids (Hanseniella sp.) are polyphagous soilborne parasites. Today, symphylid populations on pineapple are monitored by observing root symptoms and the presence of symphylids at the bottom of basal leaves. The authors developed a reliable method with a bait and trap device to monitor symphylid populations in pineapple or fallow crops. The spatial distribution of the symphylid populations was evaluated using the variance/mean ratios and spatial analyses based on Moran's and Geary's indices. The method has been tested to monitor symphylid populations at different developmental stages of pineapple. RESULTS: Adding potato baits to the soil samples increased the trapping efficiency of symphylids when compared with ‘soil only’ and ‘bait only’ methods. The handling of the samples is also facilitated by the new device. Results showed that the vertical distribution of symphylids may be uniform deeply inside the soil profile under pineapple, up to 50 cm. Results showed that symphylid populations are highly aggregated, showing a spot area about 4–6 m wide for their development. CONCLUSION: The new method allows better and easier evaluation of symphylid populations. It may be very useful in the evaluation of new IPM methods to control symphylids under pineapple. Copyright © 2011 Society of Chemical Industry     

Chemical compositions of the essential oils of the aerial parts of Chamaemelum mixtum (L.) Alloni

The chemical compositions of the aerial parts essential oils of Chamaemelum mixtum (L.) Alloni from Corsica and Sardinia were investigated employing gas chromatography and gas chromatography-mass spectrometry (GC-MS). The structure of (Z)-heptadeca-9,16-dien-7-one, a natural compound not previously described, was elucidated by GC-MS (electron impact and chemical ionization) and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy. The variation in C. mixtum essential oil was studied, and statistical analysis showed the clustering of oil samples into three groups according to the amount of oxygenated compounds; these groups correlated to the harvest area. The strong biological activity of the oxygenated fraction (minimum inhibitory concentration of <0.1 mg/mL) of the Corsican oil against Candida albicans , Citrobacter frendii , Enterococcus faecalis , Escherichia coli , Klebsiella pneumoniae , Listeria monocytogenes , and Staphyllococcus aureus can be attributed to the presence of irregular monoterpene alcohols and (Z)-heptadeca-9,16-dien-7-one.     

Long‐term survival with stereotactic radiotherapy for imaging‐diagnosed pituitary tumors in dogs

Published studies on the use of stereotactic radiotherapy for dogs with pituitary tumors are limited. This retrospective observational study describes results of stereotactic radiotherapy for 45 dogs with imaging‐diagnosed pituitary tumors. All dogs were treated at a single hospital during the period of December 2009–2015. The stereotactic radiotherapy was delivered in one 15 Gray (Gy) fraction or in three 8 Gy fractions. At the time of analysis, 41 dogs were deceased. Four were alive and censored from all survival analyses; one dog received 8 Gy every other day and was removed from protocol analyses. The median overall survival from first treatment was 311 days (95% confidence interval 226–410 days [range 1–2134 days]). Thirty‐two dogs received 15 Gy (median overall survival 311 days; 95% confidence interval [range 221–427 days]), and 12 received 24 Gy on three consecutive days (median overall survival 245 days, 95% confidence interval [range 2–626 days]). Twenty‐nine dogs had hyperadrenocorticism (median overall survival 245 days), while 16 had nonfunctional masses (median overall survival 626 days). Clinical improvement was reported in 37/45 cases. Presumptive signs of acute adverse effects within 4 months of stereotactic radiotherapy were noted in 10/45, and most had improvement spontaneously or with steroids. Late effects versus tumor progression were not discernable, but posttreatment blindness (2), hypernatremia (2), and progressive neurological signs (31) were reported. There was no statistical difference in median overall survival for different protocols. Patients with nonfunctional masses had longer median overall survival than those with hyperadrenocorticism (P = 0.0003). Survival outcomes with stereotactic radiotherapy were shorter than those previously reported with definitive radiation, especially for dogs with hyperadrenocorticism.     

Light-scattering study of the structure of aggregates and gels formed by heat-denatured whey protein isolate and beta-lactoglobulin at neutral pH

The structure of aggregates and gels formed by heat-denatured whey protein isolate (WPI) has been studied at pH 7 and different ionic strengths using light scattering and turbidimetry. The results were compared with those obtained for pure beta-lactoglobulin (beta-Lg). WPI aggregates were found to have the same self-similar structure as pure beta-Lg aggregates. WPI formed gels above a critical concentration that varied from close to 100 g/L in the absence of added salt to about 10 g/L at 0.2 M NaCl. At low ionic strength (<0.05 M NaCl) homogeneous transparent gels were formed, while at higher ionic strength the gels became turbid but had the same self-similar structure as reported earlier for pure beta-Lg. The length scale characterizing the heterogeneity of the gels increased exponentially with increasing NaCl concentration for both WPI and pure beta-Lg, but the increase was steeper for the former.     

Individual heterogeneity in erythrocyte susceptibility to Babesia divergens is a critical factor for the outcome of experimental spleen-intact sheep infections

Susceptibility of sheep erythrocytes to Babesia divergens was investigated in vitro and a high inter-individual variability in their ability to support parasite population development was demonstrated, with some individuals having refractory red blood cells (RBC). As neither changes in growth conditions nor the use of different B. divergens strains influenced the level of susceptibility, the main factor postulated for this variability is the erythrocyte itself. Sheep therefore represent an excellent in vitro model to study the parasite-erythrocyte interaction. In addition, the existence of refractory RBC should help in the identification of the erythrocyte components required for B. divergens development. Experimental infections were carried out on spleen-intact sheep characterized by refractory or fully susceptible erythrocyte types. These differences translated into the successful infection of only those animals with susceptible erythrocytes: infected animals showed no clinical signs, but maintained an asymptomatic persistent infection, as usually observed in the natural bovine host. Sheep therefore represent model organisms that can allow us to study interactions between B. divergens and its vertebrate host at different levels of biological organisation, from the target cell to the intact animal, and represent an experimental infection model of concomitant immunity. Only a low percentage (13%) of the sheep population tested possessed susceptible erythrocytes and the potential role of sheep as a natural host or reservoir of B. divergens is discussed.     

Comparative long-term efficacy of ivermectin and moxidectin over winter in Canadian horses treated at removal from pastures for winter housing

The impact of a late fall treatment on the spring rise of fecal egg counts was evaluated in a controlled study with Canadian horses treated with 2 different dewormers immediately after removal from pasture for winter housing. The horses were stabled until the end of the trial period. Seventeen weanlings, 20 yearlings, and 15 2-year-old horses located in Ontario, which were presumed to be naturally infected with cyathostomins after pasture grazing, were randomly allocated to either a group treated with 0.4 mg/kg of moxidectin and 2.5 mg/kg of praziquantel or a group treated with 0.2 mg/kg of ivermectin and 1.5 mg/kg of praziquantel. Three weeks after treatment, all strongyle fecal egg counts were reduced to zero for both treatment groups. However, at 5 months post-treatment, mean geometric fecal egg counts were statistically higher for the yearlings and 2-year-old horses treated with ivermectin than for the yearlings and 2-year-old horses treated with moxidectin (P < 0.0001).