Studies on Survival of Horse Oocytes After Rapid-i Method Vitrification

The aim of the present study was to determine the vitality and developmental competence of equine oocytes after in vitro maturation (IVM) and vitrified by Rapid-i method. In experiment 1, oocytes after IVM were vitrified using media: EquiPro VitKit (group 1) or medium containing 18% Ficoll, 40% ethylene glycol, and 0.3 M sucrose (group 2). For evaluation of toxicity effect, oocytes were exposed to media without a plug to liquid nitrogen. To evaluate viability, oocytes were stained with fluorescein diacetate and ethidium bromide. In experiment 2, oocytes after IVM and vitrification were activated by 7.5 μM ionomicin in TCM 199 (5 minutes) combined with 2 mM 6-DMAP in TCM 199 with 10% fetal bovine serum (4.5 hours). Survival rate was: 63% in group 1 (n = 54), 55% in group 2 (n = 69), and 73.2% (n = 56) in the control group. After parthenogenetic activation, 10.2% (n = 49) of 2–4 blastomeres were observed. This percentage was lower than in the nonvitrified group: 38.5% (n = 53).     

Estimation of the number of exposed people during highly pathogenic avian influenza virus outbreaks in EU/EEA countries,October 2016–September 2018

We estimated that more than 11,000 people were exposed to highly pathogenic avian influenza viruses in EU/EEA countries over the outbreak period October 2016–September 2018 by cross‐linking data submitted by Member States to European Food Safety Authority and EMPRES‐i. A stronger framework for collecting human exposure data is required.     

The production and distribution of IL-6 and TNF-α in subcutaneous adipose tissue and their correlation with serum concentrations in Welsh ponies with equine metabolic syndrome

A main symptom of equine metabolic syndrome (EMS) in ponies is pathological obesity characterized by abnormal accumulation of fat deposits and inflammation. In this study, we analyzed the expression of two pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), in subcutaneous adipose tissue and the correlation with serum concentrations in peripheral blood of Welsh ponies. Based on clinical examination findings, the animals were divided into two groups: ponies affected with EMS (n = 8) and obese ponies (n = 8). The adipose tissue was examined using immunohistochemical analysis while concentrations IL-6 and TNF-α were measured using enzyme-linked immunosorbent assays (ELISAs). Additionally, histological characterization of the adipose tissue was performed. The results obtained showed that IL-6 expression in adipose tissue biopsies derived from animals with EMS was enhanced while TNF-α levels of both groups were comparable. Compared to the obese ponies, EMS animals also had significantly elevated levels of serum IL-6 and TNF-α. Histological analysis revealed macrophage infiltration and fibrosis in adipose tissue preparations from the EMS group. These data suggest that IL-6 may play a key role in the course of EMS in Welsh ponies. Our findings also demonstrated that analysis of pro-inflammatory cytokines levels in serum may serve as an additional tool for diagnosing EMS.     

The effect of the source and dosage of dietary Cu on redox status in rat tissues

The aim of this experiment was to investigate whether the amount of Cu added to the diet of rats can be reduced without adversely affecting the antioxidant status of tissues and growth, and whether copper nanoparticles can be used for this purpose. For four weeks, four experimental groups of rats were fed diets with two dosages of added Cu (standard—6.5 or 3.25 mg/kg) in two forms (standard—CuCO₃ or copper nanoparticles). Replacing the CuCO₃ supplement with CuNPs resulted in a decreased lung weight and an increased Cu content in brain, kidney and lung, intensification of lipid peroxidation processes, and weakened antioxidant defence in the lungs and kidneys. This treatment also reduced the Cu content in heart, level of lipid oxidation in the liver and testes and improved antioxidant defence in the brain. Reducing the addition of Cu to the diet from 6.5 to 3.25 mg/kg reduced lung weight and increased lipid peroxidation in the liver, heart and lungs, and also weakened antioxidant defence in the lungs and testes. This treatment also weakened the lipid peroxidation process in the spleen, small intestine and brain and strengthened the antioxidant defence of the brain and kidneys. In conclusion, replacing CuCO₃ with CuNPs and reducing the level of Cu in the diet of rats has a particularly unfavourable effect on the respiratory system, causing adverse changes in the lungs. However, these treatments have a clearly positive effect on the redox status of the liver and brain.     

Cyprinid herpesvirus 3: an interesting virus for applied and fundamental research

Cyprinid herpesvirus 3 (CyHV-3), a member of the family Alloherpesviridae is the causative agent of a lethal, highly contagious and notifiable disease in common and koi carp. The economic importance of common and koi carp industries together with the rapid spread of CyHV-3 worldwide, explain why this virus became soon after its isolation in the 1990s a subject of applied research. In addition to its economic importance, an increasing number of fundamental studies demonstrated that CyHV-3 is an original and interesting subject for fundamental research. In this review, we summarized recent advances in CyHV-3 research with a special interest for studies related to host-virus interactions.     

The influence of \upbeta-hydroxy- \upbeta-methylbutyrate (HMB) on cell-mediated immunity in tench Tinca tinca (L.): in vitro and in vivo study

$\upbeta$ -Hydroxy- $\upbeta$ -methyl butyrate (HMB) has been shown to counteract many of the negative effects of intensive fish production methods and results in increased growth and protection against diseases. In the present study, the in vitro and in vivo effect of HMB on the immunocompetence cell activity in tench Tinca tinca (L.) was examined. In the in vitro study pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing 0, 0.1, 1, 5, 10, 25, 50 or $100\,\upmu$ g HMB cm^?3 of medium. The effects of HMB on the respiratory burst activity (RBA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by concanavalin A (ConA) or lipopolysaccharide (LPS) were examined. The in vitro study showed that HMB increased the RBA and PKA of the phagocytes, compared to the medium over that of cells grown without HMB. Lymphocyte proliferation stimulated by ConA and LPS was also increased approximately when HMB was added to the culture medium at concentrations between 10 and $100\,\upmu$ g HMB cm^?3. In the in vivo study fish were fed daily with pellets containing HMB at doses 0, 10, 25 and 50 mg kg^?1 body weight day^?1. The study showed that HMB statistically increased the RBA and PKA and highest activity at 50 mg kg^?1 body weight day^?1 were observed. Also lymphocyte proliferation stimulated by ConA and LPS were maximally stimulated at dose 50 mg kg^?1 body weight day^?1. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in tench through increased cell proliferation and functionality.     

Electrochemically mediated atom transfer radical polymerization

Atom transfer radical polymerization is a versatile technique for exerting precise control over polymer molecular weights, molecular weight distributions, and complex architectures. Here, we show that an externally applied electrochemical potential can reversibly activate the copper catalyst for this process by a one-electron reduction of an initially added air-stable cupric species (Cu(II)/Ligand). Modulation of polymerization kinetics is thereby tunable in real time by varying the magnitude of applied potential. Application of multistep intermittent potentials successfully triggers initiation of polymerization and subsequently toggles the polymerization between dormant and active states in a living manner. Catalyst concentrations down to 50 parts per million are demonstrated to maintain polymerization control manifested in linear first-order kinetics, a linear increase in polymer molecular weight with monomer conversion, and narrow polymer molecular weight distributions over a range of applied potentials.     

A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.     

Mycoplasma hominis ssp. associated endocarditis with myocardial necrosis in an alpaca (Vicugna pacos) in Manitoba in 2011

Severe endocarditis with myonecrosis, moderate to severe pleural and pericardial effusions, and mild ascites were found on necropsy in 3 alpacas. Mycoplasma hominis ssp. was detected on polymerase chain reaction (PCR) of fresh affected endocardial tissue in 1 alpaca.