Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis

Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster infections to sustain ongoing immunity.     

Antepartal insulin-like growth factor concentrations indicating differences in the metabolic adaptive capacity of dairy cows

Cows with different Insulin-like Growth Factor-I (IGF-I) concentrations showed comparable expression levels of hepatic growth hormone receptor (GHR). Suppressor of cytokine signaling 2 (SOCS2), could be responsible for additional inhibition of the GHR signal cascade. The aims were to monitor cows with high or low antepartal IGF-I concentrations (IGF-I^high or IGF-I^low), evaluate the interrelationships of endocrine endpoints, and measure hepatic SOCS2 expression. Dairy cows (n = 20) were selected (240 to 254 days after artificial insemination (AI)). Blood samples were drawn daily (day -17 until calving) and IGF-I, GH, insulin, thyroid hormones, estradiol, and progesterone concentrations were measured. Liver biopsies were taken (day 264 ± 1 after AI and postpartum) to measure mRNA expression (IGF-I, IGFBP-2, IGFBP-3, IGFBP-4, acid labile subunit (ALS), SOCS2, deiodinase1, GHR1A). IGF-I concentrations in the two groups were different (p < 0.0001). However, GH concentrations and GHR1A mRNA expression were comparable (p > 0.05). Thyroxine levels and ALS expression were higher in the IGF-I^high cows compared to IGF-I^low cows. Estradiol concentration tended to be greater in the IGF-I^low group (p = 0.06). It was hypothesized that low IGF-I levels are associated with enhanced SOCS2 expression although this could not be decisively confirmed by the present study.     

The complex folding network of single calmodulin molecules

Direct observation of the detailed conformational fluctuations of a single protein molecule en route to its folded state has so far been realized only in silico. We have used single-molecule force spectroscopy to study the folding transitions of single calmodulin molecules. High-resolution optical tweezers assays in combination with hidden Markov analysis reveal a complex network of on- and off-pathway intermediates. Cooperative and anticooperative interactions across domain boundaries can be observed directly. The folding network involves four intermediates. Two off-pathway intermediates exhibit non-native interdomain interactions and compete with the ultrafast productive folding pathway.     

Efficient inhibition of the Alzheimer's disease beta-secretase by membrane targeting

beta-Secretase plays a critical role in beta-amyloid formation and thus provides a therapeutic target for Alzheimer's disease. Inhibitor design has usually focused on active-site binding, neglecting the subcellular localization of active enzyme. We have addressed this issue by synthesizing a membrane-anchored version of a beta-secretase transition-state inhibitor by linking it to a sterol moiety. Thus, we targeted the inhibitor to active beta-secretase found in endosomes and also reduced the dimensionality of the inhibitor, increasing its local membrane concentration. This inhibitor reduced enzyme activity much more efficiently than did the free inhibitor in cultured cells and in vivo. In addition to effectively targeting beta-secretase, this strategy could also be used in designing potent drugs against other membrane protein targets.     

Bryde's whales in the North-East Atlantic: New insights on site fidelity and connectivity between oceanic archipelagos

1. The conservation of marine megafauna presents numerous difficulties owing to their high mobility over difficult-to-access oceanic areas that impairs the collection of basic, but essential, biological information.
2. The Bryde's whale (Balaenoptera edeni) is one of the most elusive species of baleen whales, and although it is known to be a seasonal visitor to several archipelagos in Macaronesia (the Azores, Madeira, and Canaries), there are no studies regarding its occurrence or geographical connectivity in this area of the Atlantic.
3. A 14-year photographic database was used to determine short-term (intra-seasonal) and long-term (inter-annual) Bryde's whale site fidelity and to estimate individual residency times in Madeira, whereas photographic catalogues from Madeira and the Canaries were compared in order to assess large-scale movements (i.e. on the scale of hundreds of kilometres).
4. In Madeira, 59 individuals were identified, 27 (45.8%) of which were recaptured. Of these, 10 individuals (37.0%) presented short-term site fidelity and 17 individuals (63.0%) presented long-term site fidelity, with a maximum recapture interval of 12 years. Lagged identification rates showed that five individuals (SE = 2) remained in the area for 32 days (SE = 108 days) before leaving and not returning during the same year. Seven individuals were seen both in Madeira and the Canaries (catalogue comprising 51 individuals), three of which were identified multiple times in both archipelagos, with a minimum of 43 days between consecutive sightings.
5. This information combined with the fact that this species is commonly sighted accompanied by calves and feeding in both archipelagos highlights the ecological importance of this area for Bryde's whales. This should be taken into consideration by policymakers when implementing conservation measures, where coordination of effort among countries is needed. This study also reinforces the value of using data from platforms of opportunity and of making photographic data open access.

     

Influence of toltrazuril treatment on parasitological parameters and health performance of piglets in the field--an Austrian experience

Porcine coccidiosis caused by Isospora suis is one of the leading causes of neonatal diarrhea in suckling piglets. Currently the only registered drug for metaphylaxis is toltrazuril. To evaluate the effect of treatment on piglets from 7 Austrian farms without and 8 Austrian farms with toltrazuril application we examined oocyst excretion (including determination of oocysts per gram of feces; OPG), diarrhea (fecal score FS 1-4 with 3 and 4 being diarrhea), and general health (health score HS 1-4 with 3 and 4 describing poor health). Both groups included farms with different levels of hygiene. Samples from 265 litters without treatment, comprising 1588 individual samples, and 1548 samples from 258 treated litters were taken twice (around the 14th and the 21st day of life, respectively), examined by autofluorescence and, if positive, by McMaster counting. In both groups animals had less diarrhea and lower health scores during the second sampling but the treated piglets were always significantly healthier and had less diarrhea. The percentage of weaned piglets was higher in treated animals although this was not significant (p=0.052). In the first round of sampling 17.8% of the individual samples from untreated piglets were positive for oocysts (with a maximum prevalence on the 12-15th day of life) while in the treated piglets only 0.4% shed oocysts p<0.001). At the second sampling only 2.1% of the untreated animals and none of treated piglets excreted I. suis (p=0.083). Positive animals shed up to 8 × 10(3)OPG. There was an increased risk for infected piglets to develop diarrhea (odds ratio, OR 4.73) and poor health (OR 5.05) in untreated piglets, and poor hygiene without disinfection was identified as a risk factor for poor health (OR 1.90), diarrhea (OR 1.42) and oocyst excretion (OR 1.73). The risk of poor health (OR 2.89) and diarrhea (OR 1.44) was also increased for piglets under poor hygienic conditions receiving toltrazuril, so both metaphylaxis of coccidiosis and good hygiene are necessary to effectively control neonatal diarrhea. The costs of treatment are considerably lower than the estimated financial production losses. Therefore, treatment is recommended for farms where clinical coccidiosis is diagnosed.     

Effect of antecedent soil moisture conditions on emissions and isotopologue distribution of N2O during denitrification

The present study determined the influence of initial moisture conditions on the production and consumption of nitrous oxide (N₂O) during denitrification and on the isotopic fingerprint of soil-emitted N₂O. Sieved arable soil was pre-incubated at two different moisture contents: pre-wet at 75% and pre-dry at 20% water-filled pore space. After wetting to 90% water-filled pore space the soils were amended with glucose (400 kg C ha⁻¹) and KNO₃ (80 kg N ha⁻¹) and incubated for 10 days under a He/O₂-atmosphere. Antecedent moisture conditions affected denitrification. N₂ + N₂O fluxes and the N₂O-to-N₂ ratio were higher in soils which were pre-incubated under dry conditions, probably because mobilization of organic C during the pre-treatment enhanced denitrification. Gaseous N fluxes showed similar time patterns of production and reduction of N₂O in both treatments, where N₂O fluxes were initially increasing and maximised 3-4 days after fertilizer application, and N₂ fluxes were delayed by 1-2 days. Time courses of δ¹⁵N^bulk-N₂O and δ¹⁸O-N₂O exhibited in both treatments increasing trends until maximum N₂ fluxes occurred, reflecting isotope fractionation during intense NO₃⁻ reduction. Later this trend slowed down in the pre-dry treatment, while δ¹⁸O-N₂O was constant and δ¹⁵N^bulk-N₂O decreased in the pre-wet treatment. We explain these time patterns by non-homogenous distribution of NO₃⁻ and denitrification activity, resulting from application of NO₃⁻ and glucose to the surface of the soil. We assume that several process zones were thus created, which affected differently the isotopic signature of N₂O and the N₂O and N₂ fluxes during the different stages of the process. We modelled the δ¹⁵N^bulk-N₂O using process rates and associated fractionation factors for the pre-treated soils, which confirmed our hypothesis. The site preference (SP) initially decreased while N₂O reduction was absent, which we could not explain by the N-flux pattern. During the subsequent increase in N₂ flux, SP and δ¹⁸O-N₂O increased concurrently, confirming that this isotope pattern is indicative for N₂O reduction to N₂. The possible effect of the antecedent moisture conditions of the soil on N₂O emissions was shown to be important.     

The Pharmacokineties of Alfentanil in Healthy Cats

Alfentanil (50 μg/kg) was administered intravenously, over 1 minute, to 6 healthy cats. Blood samples were collected from a preplaced arterial catheter at 0.625, 1.25, 2.5, 5, 10, 15, 20, 30, 45, 60, 90, 120, 180, 240, 360, and 480 minutes after the end of the alfentanil injection. A radio-immunoassay technique was used to measure alfentanil concentrations in plasma obtained from these samples. Arterial blood pressure was measured at 0.625, 1.25, 2.5, 5, 10, 15, 20, and 30 minutes and pH and blood gas measurements were carried out at 5, 10 and 30 minutes after the alfentanil. Analgesia, tested by placing a clamp on the base of the tail for 5 seconds, was assessed at each blood sampling time until analgesia was absent. The cats were observed for behavioural changes at each sample time. Alfentanil caused a transient increase in blood pressure and respiratory and metabolic acidosis. Following alfentanil all cats became excited and showed pupillary dilation. Analgesia was present for 21.7 ± 14 minutes. The plasma concentration-time data for 5 cats were best described by a 3 compartment open model with the curve fitting the equation: C_to= 162.6e^-0.2062t+ 60.le^-0.041t+ 13.2e^-0.0062t The harmonic means for the half lives of the rapid distribution, the slow distribution and the elimination phases were 4.12, 18.8, 119.2 minutes respectively.