Comparative external morphology of cetacean spermatozoa

SUMMARY: To compare size and morphology of spermatozoa in cetaceans, sperm and epididymis samples were collected from 10 species in four families and spermatozoa were observed with phase-contrast and scanning electron microscopes. According to the average total length of the spermatozoa, the 10 species examined were classified into the following four groups in order of increasing size: (i) Baird's beaked (Ziphiidae) and Bryde's whales (Balaenopteridae); (ii) Dall's and finless porpoises (Phocoenidae); (iii) common, bottlenose, and Pacific white-sided dolphins (Delphinidae); and (iv) killer and short-finned pilot whales, and Risso's dolphin (Delphinidae). Spermatozoa head length of Bryde's whale and the finless porpoise were shorter than those of the other species. Spermatozoa head width was widest in the killer whale and thinnest in the Baird's beaked whale. The lateral aspects of sperm heads from the 10 species were characterized as the 'anterior region of the sperm head is thin and flat while the posterior region is thick.' The dorsal aspects of sperm heads were 'paddle-shaped' in Bryde's whales, 'bowling pin-shaped' in Baird's beaked whales, 'Japanese fan-shaped' in killer whales, an 'elongated ellipsoid shape' in Delphinidae except for killer whales, and 'ellipsoid shaped' in Phocoenidae. Size and morphology of the spermatozoa showed interspecific differences among the 10 species examined, which correspond to cetacean taxonomic classification.     

Ultrasonic observations on the turnover of ovarian follicular cysts and associated changes of plasma LH, FSH, progesterone and oestradiol-17β in cows

Changes in the diameters of individual follicular structures on ovaries were measured by transrectal ultrasonography for 29 to 40 days and the plasma concentrations of luteinising hormone (LH), follicle-stimulating hormone (FSH), progesterone and oestradiol 17β were determined in four cows with ovarian cysts. When these structures decreased in size, new follicular structures appeared and developed into cysts. Progesterone concentrations in plasma were below 1·0 ng ml⁻¹ during the experimental periods. Plasma concentrations of oestradiol-17β fluctuated. The mean concentration of oestradiol-17β in plasma differed (P<0·01) depending on the stage of the cyst. No preovulatory surges of LH were detected during the developmental stage of the cysts.     

Improved survival of vitrified in vivo-derived porcine embryos

An efficient cryopreservation protocol for porcine morulae was investigated with three types of vitrification having different cooling rates (Exp. 1). Survival of embryos vitrified after removal of cytoplasmic lipid droplets was also examined by means of the minimum volume cooling (MVC) method (Exp. 2). In Exp. 1, the morula stage embryos were vitrified with a 0.25 ml plastic straw (ST-method), gel loading tip (GLT-method) and the MVC-method, respectively, and stored in liquid nitrogen after which they were warmed in sucrose solutions with cryoprotectants being subsequently removed in a stepwise manner. In Exp. 2, morulae were centrifuged with 7.5 microg/ml cytocharasin B at 12000 x g for 20 min to polarize the cytoplasmic lipid droplets that were then removed from the embryos by micromanipulation (delipation). Both those delipated at the morula stage and the intact embryos at the morula to blastocyst stages were vitrified by the MVC-method. In vitro survival of the vitrified embryos was assessed in both experiments by culturing in NCSU-23 + 10% FCS for 48 h. In vitro developments of vitrified embryos after warming to blastocysts were 20% (6/30) for the ST-method, 39% (18/46) for the GLT-method, and 60% (26/43) for the MVC-method. Embryo survival was further improved by vitrification after delipation (95%, 35/37) compared to intact vitrified morulae (24/42, 57%, P<0.001) and blastocysts (23/31, 74%, P<0.05). Moreover, the number of cells in blastocysts (92 +/- 25) derived from the delipated-vitrified morulae was comparable to those derived from intact control non-vitrified embryos (103 +/- 31). Our results demonstrate that vitrified porcine morulae have the highest survival when using the MVC-method in conjunction with delipation.     

Is the decline of birds and amphibians in a steppe lake of northern Patagonia a consequence of limnological changes following fish introduction?

• 1. Laguna Blanca, in Laguna Blanca National Park, is a lake in Patagonia which has been designated as a Ramsar site since 1971 because of bird diversity and abundance and importance for nesting, particularly for the black neck swan, Cygnus melanocoryphus. It is also valued for its populations of endemic amphibians, Atelognathus patagonicus and Atelognathus praebasalticus.
• 2. Avian and amphibian populations have decreased dramatically in recent years. Percichthys colhuapiensis, Percichthys trucha (Pisces, Percichthyidae), Salmo trutta and Oncorhynchus mykiss (Pisces, Salmonidae) were introduced into Laguna Blanca in 1965. Since 1986, no Atelognathus frogs have been found. The abundance of swans and coots, which are strongly associated with macrophytes for nesting and feeding, has diminished drastically, whereas piscivorous birds have increased.
• 3. The fishless condition of some neighbouring small lakes with abundant pond weeds, aquatic birds and endemic amphibians, was assessed in order to compare the physical and chemical characteristics and the quantitative composition of the benthos among lakes. Fish presence at Laguna Blanca and its absence at El Burro, Antiñir and Jabón lakes, were confirmed. Compared with previous results, it seems that the abundance of Amphipoda (Hyalella), Copepoda and Cladocera at Laguna Blanca has decreased, while Acari has increased and Notostraca has disappeared. Water transparency has diminished in Laguna Blanca and now is lower than that of fishless lakes.
• 4. P. colhuapiensis were captured only in Laguna Blanca, with the highest captures in the littoral zone. The population shows rapid individual growth in the early years and an absence of fish older than 6 years. The preponderance of benthos and the presence of macrophytes in the gut contents of adult P. colhuapiensis appear to indicate that they are important consumers of these resources.
• 5. This paper concludes that fish introduction in Laguna Blanca led to a complex trophic cascade effect (fish predation on tadpoles, fish competition for avian food, bottom disturbance, zooplankton reduction) producing deleterious effects on the amphibian and bird populations.

Copyright © 2006 John Wiley & Sons, Ltd.     

Use of image analysis to estimate anthocyanin and UV-excited fluorescent phenolic compound levels in strawberry fruit

Strawberry is rich in anthocyanins, which are responsible for the red color, and contains several colorless phenolic compounds. Among the colorless phenolic compounds, some, such as hydroxycinammic acid derivatives, emit blue-green fluorescence when excited with ultraviolet (UV) light. Here, we investigated the effectiveness of image analyses for estimating the levels of anthocyanins and UV-excited fluorescent phenolic compounds in fruit. The fruit skin and cut surface of 12 cultivars were photographed under visible and UV light conditions; colors were evaluated based on the color components of images. The levels of anthocyanins and UV-excited fluorescent compounds in each fruit were also evaluated by spectrophotometric and high performance liquid chromatography (HPLC) analyses, respectively and relationships between these levels and the image data were investigated. Red depth of the fruits differed greatly among the cultivars and anthocyanin content was well estimated based on the color values of the cut surface images. Strong UV-excited fluorescence was observed on the cut surfaces of several cultivars, and the grayscale values of the UV-excited fluorescence images were markedly correlated with the levels of those fluorescent compounds as evaluated by HPLC analysis. These results indicate that image analyses can select promising genotypes rich in anthocyanins and fluorescent phenolic compounds.     

Embryo collection from pigs post‐pseudopregnancy induced by estradiol dipropionate

We aimed to define whether embryo collection carried out after pseudopregnancy was of similar outcome and quality as after artificial abortion. To induce pseudopregnancy, 30 gilts or sows were given 20 mg intramuscular estradiol dipropionate (EDP) 10–11 days after the onset of estrus. Ten additional pigs were inseminated artificially at natural estrus as a control group. Prostaglandin F_2α (PGF_2α) was administered twice with a 24 hr interval beginning 15, 20, or 25 days after EDP‐treatment (n = 10 per group) or between 23 and 39 days after artificial insemination in control pigs. Following this, all pigs were given 1,000 IU equine chorionic gonadotropin and 500 IU human chorionic gonadotropin (hCG) and then inseminated. Embryos were recovered 6 or 7 days after hCG treatment and outcome was recorded. There was no significant difference in the number of normal embryos collected from the pigs with PGF_2α initiated at different time points or from the control group. Embryonic developmental stages 7 days after hCG treatment also did not differ among groups. These results indicate that the use of EDP to induce pseudopregnancy, followed by PGF_2α administration to synchronize estrus for subsequent embryo harvest, is a suitable alternative to the artificial abortion method.