Activin‐A receptor expression patterns in prepubertal goat oocytes and derived embryos

This study examines the presence of activin IIA and IIB receptors (ActR‐IIA and ActR‐IIB) by Western blotting and immunocytochemistry in immature and IVM‐oocytes, 2 to 8‐cells embryos and blastocysts from prepubertal goats. Western blotting revealed that activin receptors are synthesized during oocyte maturation and embryo development. In the immunocytochemistry experiments, no immunostaining for either receptor was detected in oocytes while both receptors were immunolabelled in all the cells of cleaved embryos. In blastocysts, while ActR‐IIA expression appeared evenly distributed in the two cell lineages, inner cell mass and trophectoderm, the ActR‐IIB immunosignal was restricted mainly to the inner cell mass. Our findings reveal the presence of activin type II receptors (ActR‐IIA and ActR‐IIB) in in vitro matured prepubertal goat oocytes and blastocyst‐stage embryos. The expression of these receptors could be a key factor in understanding differences between competent and incompetent oocytes.     

Isolation and characterisation of local strains of Chlamydophila abortus (Chlamydia psittaci serotype 1) from Tunisia

Chlamydiosis is one of the major diseases that can lead to abortion in ewes. Since 1997, in 5 regions of Tunisia, Chlamydia-related abortions have been reported in 15 sheep and goat flocks. One hundred and sixty-six sera and 50 vaginal swab samples were collected from adult ewes. Chlamydial antigens were detected in 29 (58%) of the vaginal swabs using Enzyme Linked Immunosorbent Assay (ELISA) while 9 (18%) were positive by cell culture. Five strains were recovered from 4 different sheep flocks. Monoclonal antibody profiles and restriction fragment length polymorphism (RFLP) analysis of the 16S-23S rRNA spacer region showed that these isolates were C. abortus. Using amplified fragment length polymorphism (AFLP), these Tunisian strains were shown to exhibit the same pattern as strains isolated in France.     

Cultivar-based fruit size in olive depends on different tissue and cellular processes throughout growth

In drupe fruits, in addition to fruit size, the proportions of mesocarp and endocarp tissues are critical objectives for fruit quality, crop production and management. The olive fruit is a typical drupe, with cultivars which show a wide range in both fruit size and the proportions of mesocarp and endocarp. Characterizing the roles of tissue and cellular processes in producing genetically based fruit size variability is necessary for crop improvement, as well as deepening our understanding of fruit developmental physiology. This study used microscope image analysis to evaluate cell number and size, the growth of mesocarp and endocarp tissues, and their developmental timing in producing fruit size among six olive cultivars with a large range of fruit size. We found that cultivar mesocarp and endocarp size increased linearly with fruit size, with larger sizes favoring an increasingly greater mesocarp/endocarp ratio. Within the mesocarp, cultivar-based fruit size related directly to cell number and was established soon after bloom by cell division rate. In spite of different cell division rates, all cultivars showed similar timing of cell division activity, with the majority of cells produced in the two months after bloom but, surprisingly, a substantial number of cells formed during the following 6 months. Cell expansion was high throughout fruit growth and an important factor in achieving final fruit size, but cell size did not differ among cultivars at any time. We can conclude that fruit size differences among olive cultivars are due at the tissue level to both mesocarp and endocarp sizes and at the cellular level to cell division throughout fruit growth. Furthermore, since cell size is consistent among cultivars in spite of variable cell division, it is likely that cultivar differences in cell expansion accompany those in cell division.     

Olive mill wastewater effects on the microbial communities as studied in the field of olive trees by analysis of fatty acid signatures

The aim of this work was to study the effects of spreading olive mill wastewater (OMW) on the soil surface of an olive grove on the soil microbial communities. Analyses of ester-linked fatty acid methyl esters (EL-FAME) were used to assess variations in the soil microbial community structure following land spreading of OMW. Our data provide evidence that agronomic application of OMW has important effects on soil microbial community. Bacteria were relatively more reduced by these treatments than fungi and actinomycetes as revealed by an increased index of fungal/bacterial FAME and actinomycetes/bacterial FAME. Specific FAME markers indicated a significant reduction in the Gram-positive bacteria. However, the relative proportion of the Gram-negative bacteria was not significantly different after agronomic application of OMW. The ratios of cyclopropyl/monoenoic precursors decreased and the total monounsaturated/total saturated fatty acids increased in the OMW amended soils, suggesting that the microbes inhabiting the control soil are more carbon limited than the OMW amended soils. The changes in the FAME pattern of the soil organisms possibly were related (i) to an altered substrate quantity, that is the availability of substrates after the treatments, (ii) the complex nature of OMW which also contains high molecular-mass recalcitrant polyphenols.     

Changes in fatty acid composition of coriander (Coriandrum sativum L.) fruit during maturation

Changes in fatty acids were studied during maturation of coriander (Coriandrum sativum L.) fruits cultivated in the North-East of Tunisia (Charfine). The fruits matured in 55 days after flowering (DAF). Oil and petroselinic acid synthesis proceeded at a steady rate up to 32 DAF. The first results showed a rapid oil accumulation started at newly formed fruits (9.6 ± 0.2%) and continued until their full maturity (26.4 ± 0.5%). During fruit maturation, fatty acid profiles varied significantly among the nine stages of maturity. At the 32th DAF, palmitoleic, gadoleic, erucic and docosahexenoic acids were not detected and petroselinic acid had a highest amount (84.8 ± 4.5%). Fruits development resulted mainly in an increase of petroselinic acid and a decrease of palmitic acid (C16:0). At full maturity, the main fatty acids were petroselinic acid (80.9 ± 5.7%), followed by linoleic (13.6 ± 2.9%), palmitic (3.6 ± 0.1%) and stearic (0.7 ± 0.1%) acids. Saturated and polyunsaturated fatty acids decreased significantly and monounsaturated fatty acids increased during maturation of coriander fruit. Coriander fruits at the first four stages of maturity have a healthy nutritional value and the last five stages were with important economic and industrial applications. Results of this study indicate that the variation in the fatty acid composition of coriander fruit during maturation may be useful in understanding the source of nutritionally and industrially important fatty acids in this fruit. Coriander fruit is potentially an important source of petroselinic acid which has numerous industrial applications.     

Viral encephalopathy and retinopathy of Dicentrarchus labrax and Sparus aurata farmed in Tunisia

Viruses belonging to the Nodaviridae family cause disease worldwide among a large number of species of marine fish, and have been described in all continents. In the present study, a total of 69 farmed Tunisian sea bass (Dicentrarchus labrax) and 24 sea bream (Sparus aurata) samples were tested monthly for the detection of betanodavirus. The virus was identified in both species using indirect immunofluorescence assays (IFAT) and RT-PCR. In addition sequence analysis of part of the coat protein gene indicated that both species were infected by highly related, but distinct, strains belonging to the RGNNV genotype. The sequence of the coat protein gene of several strains was identical but up to 9 different sequences were detected in a single farm. In addition, viral sequences obtained from fish that were held at lower temperature (<20°C) were distinct from the rest of the sequences.     

Developmental Competence and Embryo Quality of Small Oocytes from Pre‐pubertal Goats Cultured in IVM Medium Supplemented with Low Level of Hormones,Insulin–Transferrin–Selenium and Ascorbic Acid

The aim of this study was to test the effect of insulin–transferrin–selenium (ITS) and L‐ascorbic acid (AA) supplementation and the hormonal level during in vitro maturation (IVM) of small oocytes from pre‐pubertal goat on the blastocyst yield and quality. Concretely, we used four maturation media: conventional IVM medium (CM), growth medium (GM: CM+ITS+AA and low level of hormones), modified CM (mCM: CM with low level of hormones) and modified GM (mGM: CM+ITS+AA and normal level of hormones). Cumulus–oocyte complexes (COCs) were classified into two categories according to oocyte diameter: <125 μm and ≥125 μm. Large oocytes were matured 24 h in CM (Treatment A). Small oocytes were matured randomly in six experimental groups: Treatment B: 24 h in CM; Treatment C: 12 h in GM and 12 h in CM; Treatment D: 24 h in mGM; Treatment E: 12 h in mGM and 12 h in CM; Treatment F: 12 h in mCM and 12 h in CM; and Treatment G: 12 h in GM and 12 h in mGM. After IVM, oocytes were fertilized and cultured for 8 days. The blastocyst quality was assessed by the survival following vitrification/warming and the mean cell number. When different maturation media were combined, the blastocyst rate did not improve. The large oocytes produced the highest blastocysts yield. However, the culture of small oocytes in GM (53.3%) enhanced the post‐warming survival of blastocysts compared to large oocytes matured in CM (35.7%). In conclusion, IVM of pre‐pubertal goat small oocytes in GM would be useful to improve the quality of in vitro‐produced blastocysts.     

Early growth habit and vigour parameters in olive seedlings

Fruit tree growth habit and vigour are important traits for orchard management and production, yet are difficult to select for in breeding programs because the parameters for their evaluation are complex, and their expression often requires sufficient plant size and development, entailing valuable time. Thus, in olive (Olea europaea L.) breeding, as for other fruit crops, it is critical to define growth habit and vigour parameters with selection potential, and to determine the earliest age at which they can be measured. Furthermore seedling growth habit traits are important in themselves, particularly in their relation to juvenile period length and management. To explore these issues we evaluated a series of new and standard growth habit parameters during the first year of growth in a population of unpruned olive seedlings originating from six different crosses. The influence of plant age on growth habit traits was determined by comparing measurements at two or three different times. Both parent genotype and plant age significantly affected the vigour and growth habit of the olive seedlings, and 9 months was the most appropriate age for evaluating seedling growth parameters. ‘Picual’, ‘Arbosana’ and ‘Sikitita’ were shown to be promising cultivars for use as genitors, because of their tendency to produce offspring with desirable growth habit traits such as high vigour, weeping habit and low lateral shoot number. From among the 17 parameters studied, based on parent influence and the absence of correlation among parameters, five were identified which best described olive seedling growth habit: Primary Shoot Top Diameter, Primary Shoot Conicity, Secondary Shoot Number, Secondary Shoot Insertion Angle and Longest Secondary Shoot Internode Length.