The effect of multigenerational diet containing genetically modified triticale on immune system in mice

The safety assessment of genetically modified (GM) food and feed is performed to identify the possible effects upon animal and human health, also the long-term, multigenerational influence upon functioning of different organs and systems, such as the immune system. In this study C57BL/6J mice were fed for five consecutive generations with pellets containing 20% of conventional triticale grain (control) vs. pellets containing 20% of the transgenic triticale grain resistant to BASTA herbicide (experimental). The F5 experimental animals showed enlarged inguinal and axillary lymph nodes, but not spleens, and increased WBC counts in blood (but within the norm for Mus musculus). Immunophenotyped cell suspensions derived from spleens, inguinal and axillaris lymph nodes and PBMCs from blood showed the significant decrease in the percentage of T cells in spleen and lymph nodes and the B cells in lymph nodes and blood of the F5 experimental mice in comparison to the control F5 mice. Immunoblotting analysis of IL-2, IL-4, IL-10, IL-12, IL- 6, IFN-gamma levels in serum showed significantly increased IL-2 levels and decreased IL-6 levels in the F5-experimental mice sera. No significant changes in the levels of IgE in sera in both mice groups were observed. The obtained results indicate that multigenerational use of feeds for rodents containing the GM-triticale leads to expansion of the B cell compartment in the secondary lymphoid organs, but it is not caused by malignant processes or the allergic response.     

Efficacy of topically applied liposome-bound tetracycline in the treatment of dry eye model

Objective To evaluate the effects of liposome‐bound tetracycline eye drops in a rabbit dry eye model evaluating their advantage of being less allergic, preservative free and prolonged action compared with other tear substitutes. Procedures New Zealand albino rabbits were equally divided into control group and dry eye induced groups. Dryness was induced in 24 eyes of 12 healthy adult male albino rabbits by instilling atropine sulfate eye drops 1% three times daily for 1 week, then animals were subdivided into four groups; group 1 (rabbits with dry eye model), groups 2, 3, and 4: rabbits with dry eye model treated for 7 days starting on 7th day of dryness induction with either tetracycline, empty liposome, or combined tetracycline with liposome as topical eye drops respectively. Schirmer (STT) test and tear break up time (TBUT) were assessed on days 0, 2, 4, 7, 9, 11, and 14. Animals were sacrificed on day 14 and histopathological examination of the cornea and conjunctiva was performed. Results Tear break up time and STT test values were significantly improved in groups 2, 3, 4 as compared with group 1. The histopathological examination showed normal cytoarchitecture of corneas and conjunctivae in groups 2, 3, 4 against the dryness effect that continued to affect the cornea and conjunctival epithelium in group 1. There was a significant improvement in the group treated with liposome‐bound tetracycline eye drops (group 4) as compared with tetracycline alone (group 2) and empty liposome (group 3). Conclusion The use of liposome encapsulated tetracycline significantly improved STT and TBUT values as well as reverse surface ocular pathology.     

Evaluation of ELISA in the serodiagnosis of bovine farcy

Mycobacterium farcinogenes is the causal agent of bovine farcy, a chronic infectious disease of zebu cattle in some parts of tropical Africa. Whole cell homologous antigen of M. farcinogenes was used in the standardization and evaluation of an enzyme linked immunosorbent assay (ELISA) for the detection of circulating antibodies against bovine farcy using sera from confirmed bovine farcy and from bovine farcy-free cattle. The cut-off optical density (OD) value was decided at 1.8 using filter 405nm after one hour of incubation at 37°C. Accordingly, 115 out of 124 (92.7%) serum samples from clinically proven bovine farcy cattle were reported sero-positive. Sera from cattle infected with M. avium and M. paratuberculosis revealed OD value <1.8, indicating the differential diagnostic ability of M. farcinogenes antigen. Our test sensitivity was 92.7% and specificity was 97%, therefore could be routinely employed to support early clinical diagnosis, epidemiological surveys and for screening animals before exportation to farcy-free regions.     

Physiological roles of inhibin in regulation of FSH secretion and follicular development during early pregnancy in goats

The aim of the current study was to clarify the physiological role of inhibin in controlling FSH secretion and follicular development during the early pregnancy in goats. Eight goats investigated sonographically on Days 19-21 (Day 0=day of mating) for pregnancy were assigned into control (n=3) and treated (n=5) groups. The ovaries of all animals were daily scanned with ultrasound for follicles 2mm or more in diameter from 1 day before to 5 days after treatment. On Day 25 postbreeding; animals received either 10 ml, of normal goat serum or antiserum against [Tyr (30)]-inhibin alpha (1-30). Jugular blood samples were collected every 6 h starting 24 h before and until 120 h after treatment. The plasma concentration of FSH increased at 6 h and remained at significantly high levels until 120 h in treated vs. control group. The plasma concentrations of estradiol showed a marked increased at 66 h, with peak levels at 120 h after treatment of antiserum. The basal concentrations of LH and the pattern of plasma concentrations of progesterone were not significantly different between the two groups. The number of medium size (3.5-5.0 mm) follicles increased considerably from Day 2, whereas small (3.5 mm or less) and large (5 mm or more) follicles increased noticeably from Day 3, as compared with pre-treatment and controls. These results clearly indicated that inhibin is a key hormone in regulation of follicular development through regulation of endogenous FSH secretion during early pregnancy in goats.     

Characteristics of Antioxidant Systems of Yellow Fraction of Red Deer's (Cervus elaphus L.) Semen During the Rutting Period

The objective of this study was to make the preliminary characterization of the antioxidant defence systems of the yellow fraction (YF) of red deer's (Cervus elaphus L.) semen during the rutting period. The semen was collected using artificial vagina (AV). The studies included spectrophotometric determination of antioxidant enzymes activities such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). We also analysed the contents of low‐molecular antioxidants such as L‐glutathione (GSH + GSSG), L‐ascorbate (ASC) and total antioxidant status (TAS). Additionally, the samples were subjected to PAGE and stained for SOD and GPx activities. It was demonstrated that the yellow fraction exhibited activities of SOD and GPx, with the highest activities in September and October. CAT activity was not detected. Staining for the SOD and GPx activities confirmed three protein bands with SOD activity and one protein band with GPx activity. The content of GSH + GSSG was similar in trials dating from October to December contrary to the content of ASC which was high in samples from September and October. The stable rate of TAS was observed during the whole rutting period. The results of this study showed that the YF of red deer semen is equipped with basic battery of antioxidant enzymes comprising SOD and GPx, with the supporting role of GSH + GSSG and ASC. Moreover, the samples obtained at the peak of the rutting period occurring from September to October had the highest enzymatic activity in comparison with remaining months of the rutting period, which contributed to the high quality of the semen by preventing it from the formation of oxidative stress during the short period of intense sexual activity of male red deer. The better understanding of the mechanisms of antioxidant defence systems in the YF of deer's semen may contribute to the potential use of this fraction in technology of wild ruminant semen preservation.     

Lettuce (Lactuca sativa): a species with a high capacity for cadmium (Cd) accumulation and growth stimulation in the presence of low Cd concentrations

Summary

Cadmium (Cd) is a metal pollutant that accumulates in cultivated soils and has detrimental consequences in terms of food safety. Lettuce (Lactuca sativa) can be characterised as having a high capacity to accumulate Cd in its tissues. An analysis of Cd tolerance and Cd accumulation was carried out using two varieties of lettuce (‘Divina’ and ‘Melina’). A wide range of CdCl₂ concentrations was used (0.0, 0.1, 0.6, 3.0, and 15.0 µM CdCl₂). The lowest concentration (0.1 µM CdCl₂) stimulated growth, while the two highest concentrations resulted in a reduction in biomass. Cadmium concentrations were found to be twice as high in roots as in shoots. ‘Divina’ displayed lower concentrations of Cd than ‘Melina’ in nearly all treatments. A strong negative correlation was observed between Cd concentration and Cd tolerance in the roots and shoots (R² > 0.87) of both ‘Melina’ and ‘Divina’. Lettuce grown in the presence of 15.0 µM CdCl₂ had leaf Cd concentrations that were 100-fold higher than the legal maximum level for vegetable products marketed for human consumption, but showed no symptoms of dehydration, chlorosis, or necrosis. This result represents an important alert for lettuce consumers and growers.     

Flowering and fruiting phenology,and physico-chemical characteristics of 2-year-old plants of six species of Opuntia from eight regions of Morocco

SUMMARY

We have described the flowering and fruiting phenology of 14 accessions of six Opuntia species grown in Morocco: O. ficus-indica, O. robusta, O. aequatorialis, O. dillenii, O. leucotricha, and O. stricta. We also determined the physicochemical characteristics of their cladodes. For each accession, we used four-to-20 plants. Two systems were used to measure phenology: the extended Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH)-scale and the National Phenology Monitoring System. Five principal stages were described: the development of vegetatively-propagated organs (vegetative buds), the development of flower buds, flowering, fruit development, and fruit maturity. The number of vegetative buds varied significantly among the 14 accessions, while only four accessions produced flower buds. After reaching their final size, fruit lengths varied from 3.40–6.40 cm, while fruit diameters varied from 1.93–3.90 cm. The average cladode number differed significantly among genotypes and varied from 1.75–7.75. There were also significant differences in cladode length (13.75–30.63 cm), width (6.25–17.33 cm), thickness (0.65–1.38 cm), fresh weight (FW; 67.50–766.00 g), and dry weight (DW; 8.75–67.15 g) between species. Water content and ash content, as well as total protein and total sugar concentrations, showed significant differences and were within the ranges of 86.67–92.04% (w/w), 12.97–22.08g 100 g^?1 DW, 4.64–11.56g 100 g^?1 DW, and 3.22–12.51 g 100 g^?1 DW, respectively. Our results will help in the development of agronomic management practices and improve the characterisation, valorisation, and use of cactus pear species grown in Morocco.     

In vitro micrografting of apical and axillary buds of cacao

This study aimed to evaluate in vitro grafting of Theobroma cacao where seedlings of the UF 677 genotype were used as the rootstock and apices or axillary buds of a Trinitarian genotype were used as scion. Three methods of grafting using scions from seedlings were evaluated. Apical grafts using apex and side grafts using apex displayed better graft success (95 and 80%, respectively). However, side grafts using axillary buds reached a greater height on average and a higher number of leaves per plant (1.76 cm and 3.72, respectively). Histological studies revealed new vascular elements at the graft union area. Side grafts with axillary buds provided the highest survival rate (82%) after the acclimatization step. A shoot of at least 1 cm with two leaves is required for plant survival after transfer to ex vitro conditions. Side grafting was carried out with axillary buds from adult trees and nursery plants. Only the grafts with buds from nursery grafted plants were successful, with a rate of 26%. Overall, side grafting with axillary buds is the most appropriate method for cacao micrografting. This method can be used for clonal propagation and for the establishment of in vivo and/or in vitro cacao germplasm collection.