Induction of CIRBP expression by cold shock on bovine cumulus–oocyte complexes

The aim of this study was to induce the cold‐inducible RNA‐binding protein (CIRBP) expression on cumulus–oocyte complexes (COCs) through exposure to a sub‐lethal cold shock and determine the effects of hypothermic temperatures during the in vitro maturation of bovine oocytes. Nuclear maturation, cortical granule redistribution and identification of cold‐inducible RNA‐binding protein (CIRBP) were assessed after 24 hr of in vitro maturation of control (38.5°C) and cold‐stressed oocytes (33.5°C). The presence of CIRBP was assessed by Western blot in COCs or denuded oocytes and their respective cumulus cells. Based on the odds ratio, cold‐stressed oocytes presented higher abnormal cytoplasmic distribution of cortical granules and nuclear maturation than the control group. Although CIRBP was detected in both control and cold‐stressed groups, cold‐stressed COCs had 2.17 times more expression of CIRBP than control COCs. However, when denuded oocytes and cumulus cells were assessed separately, CIRBP only was detected in cumulus cells in both groups. In conclusion, cold shock induced CIRBP expression, but it negatively affected nuclear maturation and cortical granule distribution of bovine oocytes. Moreover, the expression of CIRBP was only identified in cumulus cells but not in oocytes.     

The effect of row spacing on alfalfa seed and forage production under irrigated Mediterranean agricultural conditions

We evaluated the effect of row spacing on seed and forage yields in alfalfa (Medicago sativa L.) using combined seed and forage production plots, in an irrigated Mediterranean agricultural environment. We compared three row distances (20, 40 and 60 cm) in two cultivars at a seed rate of 25 kg ha^?1 over a period of 4 years. We measured the seed yield and yield components as well as forage production. The 4‐year average revealed an overall clean seed yield of 367 kg ha^?1 declining over time, without significant differences in row distances. The number of shoots per square metre differed significantly as a function of row spacing, with averages of 357, 226 and 172 shoots m^?2 for row widths of 20, 40 and 60 cm respectively. The row spacing did not affect the number of racemes per shoot (23·1), the number of pods per raceme (7·2), the number of seeds per pod (6·5) or the thousand seed weight (1·667 g). The average forage production was 20·1, 18·5 and 17·9 Mg DM ha^?1 for row distances of 20, 40 and 60 cm, respectively, with higher yields associated with smaller row distances. Our data demonstrate that row spacing >20 cm does not confer any advantage to the production of alfalfa seed under irrigated Mediterranean agricultural conditions and that close spacing can significantly increase forage yields in the same environment.     

Bacteria associated with winter mortalities in laboratory-reared common dentex (Dentex dentex L.)

A pathological study was conducted on laboratory‐reared juvenile common dentex (Dentex dentex) suffering trickling and continuous mortalities. During a 3‐month period (October–December), water temperature, clinical signs and mortalities were recorded. Moribund or dead fish were examined for bacteria and parasites. Bacteria were isolated from head kidney and external ulcers, and samples from the gills, intestine, stomach, trunk kidney, gall bladder and liver were taken for histology. Cumulative mortality reached 73%, and 80% of fish examined were positive for bacteria (102 isolates). Vibrio splendidus was the most prevalent in head kidney (59.7%) and ulcers (88.9%), and it was frequently isolated as pure culture (74.7% and 100% respectively). This is the first report of this bacterium in association with mortality in common dentex. Vibrio scophthalmi was the second most prevalent bacterium (29.2%) and accounted for 63.6% of the mixed infections with V. splendidus, V. harveyi, Pseudoalteromonas spp. and other species were rarely isolated. No parasites were found in histological sections. Bacteria were frequently observed in the lumen of the intestine and stomach, destroying the epithelium. A cellular reaction was suggested by the high numbers of rodlet cells (RC) in the intestinal epithelium, and the abundance of eosinophilic granular cells (EGC) in the intestinal lamina propria. In the stomach, vacuolized cells containing unidentified debris, sometimes of crystalloid appearance, were very common. The high prevalence of V. splendidus in pure cultures and the absence of other aetiological agents suggest that V. splendidus is substantially involved in the registered mortalities, although the implication of V. scophthalmi and even of some nutritional factors cannot be discounted.