The study of developmental capacity of vitrified mouse blastocysts in different straws after transfer to mouse pseudo pregnant

Vitrification is the commonly used method for long-term storage of pre-implantation mammalian embryos. It is an essential part of assisted reproductive technologies. The re-expansion rate, pregnancy and birth rate of vitrified blastocysts using CPS were compared with OPS and Conventional Straw. Female NMRI mice were injected with Gonadotrophins in order induce them for super ovulation. At that time the mice were sacrified by cervical dislocation and dissected of mouse abdomen. The uterine horns were existed blastocysts were collected in PBS and randomly allocated to four groups: vitrification in CPS, conventional straw, OPS and untreated controls. The vitrification solution was EFS40%. After storage for 1 month in liquid nitrogen, the blastocysts were thawed in 0.5 M sucrose for in vitro culture in M16 medium. After 6 h of culture, the numbers of expanded blastocysts was recorded and ready for transfer to uterus of pseudo pregnant mouse. The re-expansion rate of the CPS group (72.1%) was significantly higher (p < 0.05) than OPS (52.55) and C.S. (38.6%) groups. The pregnancy (70%) and birth rate (45%) of blastocysts in CPS were similar to those of fresh blastocysts (80% and 45.5%) and the pregnancy (10%) and birth rate (5.1%) in Conventional Straws lower than OPS (20 and 7.5%), but were not significantly different. Mouse blastocysts vitrified using CPS had a better result compared with OPS and Conventional Straw. The value of CPS for vitrification of blastocysts may also merit investigation.     

Establishment of a model for chemoattraction of <Emphasis Type="Italic">Symbiodinium</Emphasis> and characterization of chemotactic compounds in <Emphasis Type="Italic">Acropora tenuis</Emphasis>

Corals harbor symbiotic dinoflagellates, Symbiodinium spp., acquired from surrounding environments. Because Symbiodinium are present at low densities in the water column, corals may attract these symbionts using chemotactic compounds. To examine whether corals contain chemotactic compounds, we established an assay to measure the chemotactic activity for Symbiodinium using an extract of the coral Acropora tenuis, a major reef-building coral in Japan. Our assay revealed that Symbiodinium strain NBRC102920 (clade A), which is taken up by juvenile A. tenuis polyps, is attracted to crude A. tenuis extracts. We found that the chemotactic compounds are water-soluble, heat-labile macromolecules and that the chemotactic activity was inhibited by N-acetyl-d-glucosamine (GlcNAc). We separated the GlcNAc-binding fraction (Fr-ActL) and identified it as the most plausible candidate for the chemoattractant, since the chemotactic activity of the crude A. tenuis extract appeared to be mainly attributable to the activity of Fr-ActL and was also inhibited by the addition of GlcNAc. These results indicate that chemoattraction is mediated via the binding of Symbiodinium to Fr-ActL.     

Fucoidan and Derived Oligo-Fucoses: Structural Features with Relevance in Competitive Inhibition of Gastrointestinal Norovirus Binding

Norovirus infections belong to the most common causes of human gastroenteritis worldwide and epidemic outbreaks are responsible for hundreds of thousands of deaths annually. In humans, noroviruses are known to bind to gastrointestinal epithelia via recognition of blood-group active mucin-type O-glycans. Considering the involvement of l-α-fucose residues in these glycans, their high valency on epithelial surfaces far surpasses the low affinity, though specific interactions of monovalent milk oligosaccharides. Based on these findings, we attempted to identify polyfucoses (fucans) with the capacity to block binding of the currently most prevalent norovirus strain GII.4 (Sydney, 2012, {"type":"entrez-nucleotide","attrs":{"text":"JX459908","term_id":"409032931"}}JX459908) to human and animal gastrointestinal mucins. We provide evidence that inhibitory effects on capsid binding are exerted in a competitive manner by α-fucosyl residues on Fucus vesiculosus fucoidan, but also on the galacto-fucan from Undaria pinnatifida and their oligo-fucose processing products. Insight into novel structural aspects of fucoidan and derived oligosaccharides from low-mass Undaria pinnatifida were revealed by GCMS and MALDI mass spectrometry. In targeting noroviral spread attenuation, this study provides first steps towards a prophylactic food additive that is produced from algal species.     

The investigation of lipid peroxidation, anti-oxidant levels and some hematological parameters in sheep naturally infested with Wohlfahrtia magnifica larvae

Wohlfahrtia magnifica is the main agent of traumatic myiasis in many central and eastern European countries as well as in the Mediterranean basin and Asian countries. The present study was aimed to investigate the effect of myiasis infestation on lipid peroxidation, antioxidant levels and several hematological parameters in the sheep naturally infested with W. magnifica larvae. A total of 38 sheep, including 19 awassi sheep naturally infested with W. magnifica larvae and 19 clinically healthy awassi sheep, were studied. The infected animals were divided into three groups depending on the number of larvae (Group 1: <50 larvae; Group 2: 50-100 larvae; Group 3: >100 larvae). In blood samples, red blood cell counts, hemoglobin, hematocrit concentration and white blood cell, neutrophil, lymphocyte, eosinophil, basophil counts, plasma malondialdehyde, erythrocyte glutathione levels and erythrocyte glutathione peroxidase activity were studied. The results revealed a marked decrease in red blood cell counts and hemoglobin concentrations along with a significant increase in white blood cell and neutrophil counts. The elevation in plasma malondialdehyde levels, a function of lipid peroxidation, established a significant difference between the control group and groups 2, 3. Decreased activity of erythrocyte glutathione peroxidase was found significant in the control group and all parasitemia. We conclude that in natural infestations with W. magnifica, as in infestation with Lucilia cuprina, there is a notable increase in inflammatory activities resulting from the movements, secretions, and toxins of the larvae and form the toxins secreted by the bacteria - which leads to an impression of anemia - and that the tissue injury results in an increase in level of free radicals in the organism.     

Chemical chaperones reduce ER stress and restore glucose homeostasis in a mouse model of type 2 diabetes

Endoplasmic reticulum (ER) stress is a key link between obesity, insulin resistance, and type 2 diabetes. Here, we provide evidence that this mechanistic link can be exploited for therapeutic purposes with orally active chemical chaperones. 4-Phenyl butyric acid and taurine-conjugated ursodeoxycholic acid alleviated ER stress in cells and whole animals. Treatment of obese and diabetic mice with these compounds resulted in normalization of hyperglycemia, restoration of systemic insulin sensitivity, resolution of fatty liver disease, and enhancement of insulin action in liver, muscle, and adipose tissues. Our results demonstrate that chemical chaperones enhance the adaptive capacity of the ER and act as potent antidiabetic modalities with potential application in the treatment of type 2 diabetes.     

Effects of astaxanthin‐rich dried cell powder from Paracoccus carotinifaciens on carotenoid composition and lipid peroxidation in skeletal muscle of broiler chickens under thermo‐neutral or realistic high temperature conditions

Thirty‐two 15‐day old broiler chicks (Chunky strain ROSS 308) were randomly divided into four treatments in a 2 × 2 factorial design. The main factors were diet (basal diet or basal diet supplemented with 0.15% astaxanthin‐rich dried cell powder (Panaferd‐P [astaxanthin 30 ppm]) and ambient temperature (thermo‐neutral [25 ± 1°C] or high [35 ± 1°C for 6 hr]). Dietary supplementation with Panaferd‐P did not affect growth performance, though high ambient temperature decreased feed intake and the weight of breast tender muscle, liver, and heart. High ambient temperature also decreased redness in both breast and leg muscles of chickens, while Panaferd‐P increased redness and yellowness of breast and leg muscles of chickens. Panaferd‐P increased Paracoccus carotinifaciens‐derived pigments (i.e., adonixanthin, astaxanthin, adonirubin, and cantaxanthin) as well as corn‐derived pigments such as zeaxanthin and lutein in breast and leg muscles. High ambient temperature increased the malondialdehyde (MDA) concentration in breast muscle, while Panaferd‐P decreased the MDA concentration in breast muscle under both temperature conditions. Our results suggest that dietary supplementation with Panaferd‐P increases muscle carotenoid content, the redness and yellowness of meat and decreases the muscle MDA concentration in broiler chickens kept under thermo‐neutral or high ambient temperature conditions.     

Investigation of OECD 301F ready biodegradability test to evaluate chemical fate in a realistic environment

The OECD 301F ready biodegradability test has been approved for use under the Japanese Chemical Substances Control Law since 2018. This test uses activated sludge obtained from a sewage treatment plant instead of the standard activated sludge used for the 301C test. In addition, the test is allowed to add an inert support or emulsifying agent, and/or to change the volume of the test medium. In this study, we first confirmed that the standard sludge had lower biodegradation activities than the sludge taken from a sewage treatment plant. Second, we showed that biodegradation percentages were increased by adding suitable amounts of silica gel or Tween 80. Third, we found that the biodegradations were accelerated by only increasing the medium volume under the conditions that concentrations of chemical, silica gel, and sludge were held constant. These findings are expected to contribute to the appropriate evaluation of chemical fate in a realistic environment.     

Detection and genotyping of Toxoplasma gondii strains isolated from birds in the southwest of Iran

1. The aim of this work was to determine the frequency of occurrence of Toxoplasma gondii and genetically analyse isolates from a number of avian hosts in the southwest of Iran (Khuzestan province). The frequency of T. gondii was determined in free-range chickens (Gallus domesticus), sparrows (Passer domesticus), pigeons (Columba livia) and starlings (Sturnus vulgaris).

2. Isolates obtained from Toxoplasma-infected birds were subjected to molecular typing by PCR-restriction fragment length polymorphism (RFLP) with sequence analysis of the GRA6 gene.

3. The results showed that 41 (16·5%) of 241 samples of avian tissue were infected with T. gondii. Sparrows were most frequently infected (17 out of 64).

4. Analysis of the GRA6 gene by PCR-RFLP and DNA sequencing revealed Type II and III T. gondii were the predominant lineage, accounting for 19·5% and 80·5% of the isolates, respectively.

5. It was concluded that the use of this PCR test facilitated the diagnosis of T. gondii in avian hosts and the GRA6 PCR-RFLP method clearly differentiated between the three different T. gondii lineages. This study showed a higher prevalence of type III compared with type II T. gondii in infected avian hosts in southwestern Iran.     

Effect of dietary fibres on small intestine histomorphology and lipid metabolism in young broiler chickens

Two experiments were conducted to determine the influence of dietary fibres on small intestine histomorphology and lipid metabolism in broilers from 1 to 21 day of age. In experiment 1, diets containing insoluble [cellulose (CEL); 2% and 4%] or soluble [carboxymethyl cellulose (CMC); 2% and 4%] fibre were fed to broilers from day 1 to 21 post‐hatch and ileal tissue was collected at day 21 of age for histological evaluation. In experiment 2, broilers diet was supplemented with 0%, 1% or 2% insoluble fibre (Arbocel) during day 7 to 21 post‐hatch and plasma and liver lipid metabolism were evaluated at day 21. In experiment 1, inclusion of CMC reduced body weight gain (BWG) and feed intake (FI) and increased feed conversion ratio (FCR) compared with others. Intestinal histomorphology was unaffected by CEL, but CMC led to an increase in crypt depth (CD) and serosa thickness and a decrease in villus height (VH), villus width (VW), VH:CD ratio and villus surface area (VSA), rather than control and CEL groups. Treatment did not affect goblet cell type. Moreover, the CMC‐fed birds had greater total goblet cell count (GCC) as compared with others. In experiment 2, fibre inclusion was associated with increases in BWG from 7 to 14 day of age and an improvement in FCR, whereas FI was not influenced by treatments. Inclusion of fibre in the diet decreased the weight of the abdominal fat and cholesterol concentrations of liver and plasma. No significant effects on fatty acid composition of liver lipid were observed by fibre supplementation. These findings suggest dietary fibre affects performance, intestinal histomorphology and lipid metabolism in young chicks, which may directly affect poultry feeding strategies.