Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

Background

The aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections.

Methods

For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 10⁹ CFU pro toto).

Results

Subcutaneous inoculation of a single dose with 2.6 × 10⁹ colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation.

Conclusions

Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.     

Enigmatic hotspot of crayfish diversity at risk: Invasive potential of non‐indigenous crayfish if introduced to New Guinea

1. The large island of New Guinea has a rich indigenous astacofauna represented by numerous parastacids from the genus Cherax. The western half of the island is part of Indonesian territory.
2. Indonesia is known to be the main exporter of ornamental crayfish globally, and certain New Guinean species are exploited as ornamentals within the international pet trade. Moreover, one non‐indigenous species has been previously recorded being cultured in Java, Indonesia. This species, the North American Procambarus clarkii, is a vector of crayfish plague, the disease that is lethal to most parastacids. This population has already tested positive for the disease.
3. As the transport of non‐indigenous crayfish within the Indonesian territory is not restricted, their introduction to New Guinea can be expected. The Indonesian market was therefore surveyed for ornamental crayfish and their environmental suitability evaluated, as represented by temperature during the drought and rainy seasons in New Guinea.
4. Four North American and one Australian species were found advertised for sale. One of them, P. clarkii, was assessed as the most damaging species, followed by other North American species. A total ban on the culture and transport of the highest risk crayfish species in Indonesia and Papua New Guinea is recommended.

     

Ecosystem services from coffee agroforestry in Central America: estimation using the CAF2021 model

The goal of sustainable coffee production requires multiple functions from agroforestry systems. Many are difficult to quantify and data are lacking, hampering the choice of shade tree species and agronomic management. Process-based modelling may help quantify ecosystem services and disservices. We introduce and apply coffee agroforestry model CAF2021 (https://doi.org/10.5281/zenodo.5862195). The model allows for complex systems with up to three shade tree species. It simulates coffee yield, timber and fruit production by shade trees, soil loss in erosion, C-sequestration, N-fixation, -emission and -leaching. To calibrate the model, we used multivariate data from 32 different treatments applied in two long-term coffee agroforestry experiments in Costa Rica and Nicaragua. Without any further calibration, the model was then applied to agroforestry systems on 89 farms in Costa Rica and 79 in Guatemala where yields had been reported previously in farmer interviews. Despite wide variation in environmental and agronomic conditions, the model explained 36% of yield variation in Costa Rica but only 15% in Guatemala. Model analysis quantified trade-offs between yield and other ecosystem services as a function of fertilisation and shading.

     

Responsible Research and Innovation Framework,the Nagoya Protocol and Other European Blue Biotechnology Strategies and Regulations: Gaps Analysis and Recommendations for Increased Knowledge in the Marine Biotechnology Community

As the quest for marine-derived compounds with pharmacological and biotechnological potential upsurges, the importance of following regulations and applying Responsible Research and Innovation (RRI) also increases. This article aims at: (1) presenting an overview of regulations and policies at the international and EU level, while demonstrating a variability in their implementation; (2) highlighting the importance of RRI in biodiscovery; and (3) identifying gaps and providing recommendations on how to improve the market acceptability and compliance of novel Blue Biotechnology compounds. This article is the result of the work of the Working Group 4 “Legal aspects, IPR and Ethics” of the COST Action CA18238 Ocean4Biotech, a network of more than 130 Marine Biotechnology scientists and practitioners from 37 countries. Three qualitative surveys (“Understanding of the Responsible Research and Innovation concept”, “Application of the Nagoya Protocol in Your Research”, and “Brief Survey about the experiences regarding the Nagoya Protocol”) indicate awareness and application gaps of RRI, the Nagoya Protocol, and the current status of EU policies relating to Blue Biotechnology. The article categorises the identified gaps into five main categories (awareness, understanding, education, implementation, and enforcement of the Nagoya Protocol) and provides recommendations for mitigating them at the European, national, and organisational level.     

Comparison of the pig and feline models for full thickness corneal transplantation

Purpose The goal of this study was to report on the advantages and limitations of the pig and feline models for experimental in vivo corneal transplantation. Methods Ten healthy domestic pigs and ten healthy cats were used. Full thickness penetrating keratoplasty was performed using autologous (eight cases), allogeneic (seven cases) or human xenogeneic (three cases) tissue. In two other cases, the inflammatory response to partial thickness trephination (without transplantation) was evaluated. Eyes were assessed daily before and after surgery by slit‐lamp, pachymetry, and tonometry. A transparency score ranging from 0 (opaque graft) to 4 (clear graft) was used, based on the slit‐lamp examination. Optical coherence tomography, histology, and electron microscopy were performed postmortem. Results In the pig, the mean (±SD) transparency score for the eight full thickness grafts was 0.88 ± 0.99, ranging from 0 to 3. In the feline model, the mean transparency score for the seven uncomplicated grafts was 3.93 ± 0.19, ranging from 3.5 to 4. Both negative controls without endothelium remained opaque at all time. Intraoperative tendency for iris incarceration into the wound, rapid corneal swelling, suture cheese wiring, and postoperative intraocular inflammation were the main factors jeopardizing the functional success of the corneal transplant in the pig model. Conclusion Suboptimal functional results were obtained after full thickness corneal transplantation in the pig model, while in the feline model, the same protocol yielded uneventful surgeries and clear transplants, with functional results similar to those achieved in human subjects.     

IGF-1 receptor signaling pathways and effects of AMPK activation on IGF-1-induced progesterone secretion in hen granulosa cells

IGF-1 plays a key role in the proliferation and differentiation of granulosa cells. However, the molecular mechanism of IGF-1 action in avian granulosa cells during follicle maturation is unclear. Here, we first studied IGF-1 receptor (IGF-1R) expression, IGF-1-induced progesterone production and some IGF-1R signaling pathways in granulosa cells from different follicles. IGF-1R (mRNA and protein) was higher in fresh or cultured granulosa cells from the largest follicles (F1 or F2) than in those from smaller follicles (F3 or F4). In vitro, IGF-1 treatment (10(-8)M, 36h) increased progesterone secretion by four-fold in mixed F3 and F4 (F3/4) granulosa cells and by 1.5-fold in F1 granulosa cells. IGF-1 (10(-8)M, 30min)-induced increases in tyrosine phosphorylation of IGF-1R beta subunit and phosphorylation of ERK were higher in F1 than in F3/4 granulosa cells. Interestingly, IGF-1 stimulation (10(-8)M, 10min) decreased the level of AMPK Thr172 phosphorylation in F1 and F3/4 granulosa cells. We have recently showed that AMPK (AMP-activated protein kinase) is a protein kinase involved in the steroidogenesis in chicken granulosa cells. We then studied the effects of AMPK activation by AICAR (5-aminoimidazole-4-carboxamide ribonucleoside), an activator of AMPK, on IGF-1-induced progesterone secretion by F3/4 and F1 granulosa cells. AICAR treatment (1mM, 36h) increased IGF-1-induced progesterone secretion, StAR protein levels and decreased ERK phosphorylation in F1 granulosa cells. Opposite data were observed in F3/4 granulosa cells. Adenovirus-mediated expression of dominant negative AMPK totally reversed the effects of AICAR on IGF-1-induced progesterone secretion, StAR protein production and ERK phosphorylation in both F3/4 and F1 granulosa cells. Thus, a variation of energy metabolism through AMPK activation could modulate differently IGF-1-induced progesterone production in F1 and F3/4 granulosa cells.     

Polycyclic Aromatic Hydrocarbon Oxidation from Concentrates Issued from an Attrition Process of Polluted Soil Using the Fenton Reagent and Permanganate

This study was conducted to determine the optimal conditions for PAH degradation from highly contaminated attrition sludge (PAC) using a Fenton process or successive permanganate (KMnO₄) oxidation and Fenton processes. The following parameters were studied to optimize the Fenton oxidation process: the amounts of reactants based on the stoichiometric oxidant demand (SOD), the reactant addition protocol and number of doses, and the solid/liquid ratio (S/L). The results showed that the following conditions were optimum: TS?=?30%, 7.5 times SOD, H₂O₂/Fe²⁺ ratio?=?10, and added five times during 60 min, which allowed the degradation of 43% of total 27 PAHs from the PAC. Successive Fenton and KMnO₄ oxidation processes were also tested. PAH degradation using a sequential Fenton process followed by KMnO₄ oxidation (or KMnO₄ followed by Fenton) was higher than for the use of Fenton or KMnO₄ treatment alone. Up to 71% of the total 27 PAHs were degraded when using a combination of both processes. It appeared that the sequential treatment is a viable method for the significant degradation of 27 PAHs from PAC (t value?>?2.77).     

Health status and risk factors associated with failure of passive transfer of immunity in newborn beef calves in Québec

Risk factors associated with failure of passive transfer of immunity (FPT) were evaluated among newborn beef calves in Québec. Physical examination was performed on calves born of a normal calving and blood samples were collected for determination of health status and measurement of serum concentration of immunoglobulin (Ig) G1. Of 225 calves, from 45 herds, 19% showed FPT (serum IgG1 concentration < 10.0 g/L). Calves born in a stanchion-stall were more likely to show FPT (OR: 10.2). Calves bottle-fed colostrum were less at risk for FPT (OR: 0.06). Calf gender, month of birth, dam parity, and dam body condition score were not associated with FPT. No association was detected between FPT and health status. Special care should be given to calves born from cows in a stanchion-stall to ensure adequate colostrum intake. Failure of passive transfer of immunity should be considered with other risk factors when investigating morbidity.     

Evidence of Anti-Proliferative Activities in Blue Mussel (Mytilus edulis) By-Products

Shellfish waste components contain significant levels of high quality protein and are therefore a potential source for biofunctional high-value peptides. The feasibility of applying a pilot scale enzymatic hydrolysis process to whole Mytilus edulis and, by fractionation, recover hydrolysates presenting a biological activity of interest, was evaluated. Fractions were tested on four immortalized cancerous cell lines: A549, BT549, HCT15 and PC3. The 50 kDa fraction, enriched in peptides, presented anti-proliferative activity with all cell lines and results suggest a bioactive molecule synergy within the fraction. At a protein concentration of 44 µg/mL, the 50 kDa fraction induced a mortality of 90% for PC3, 89% for A549, 85% for HCT15 and of 81% for BT549 cell lines. At the low protein concentration of only 11 µg/mL the 50 kDa fraction still entails a cell mortality of 76% for A549 and 87% for PC3 cell lines. The 50 kDa fraction contains 56% of proteins, 3% of lipids and 6% of minerals on a dry weight basis and the lowest levels detected of taurine and methionine and highest levels of threonine, proline and glycine amino acids. The enzymatic hydrolysis process suggests that Mytilus edulis by-products should be viewed as high-valued products with strong potential as anti-proliferative agent and promising active ingredients in functional foods.