In vitro Capacitation of Ejaculated Rabbit Spermatozoa

Prior to fertilization mammalian spermatozoa undergo physiological changes in the female reproductive tract. These changes are collectively known as capacitation. In essence capacitation is a further differentiation that the sperm cell must acquire beyond the maturational changes that it undergoes in the epididymis. Although capacitation in vitro has been easily achieved in rodents, its accomplishment in the rabbit is inconsistent and difficult. We report here successful in vitro capacitation of ejaculated rabbit sperm, assessed by in vitro and in vivo fertilization of rabbit ova. Sperm were used from pooled ejaculates collected from bucks of proven fertility since sperm collected from individual bucks resulted in significant differences in fertilization levels. Conditions favoring in vitro capacitation were: (1) extended incubation time of 12 h, (2) addition of 20% heated rabbit serum to incubation medium, and (3) an atmosphere of 5% CO2, 8% O2, and 87% N2 during incubation. In vitro capacitation of sperm under these conditions resulted in 67% fertilization compared to 89% for control sperm capacitated for 13 h in the uterus.     

The Cell Cycle in Crithidia fasciculata. Temporal Relationships between Synthesis of Deoxyribonucleic Acid in the Nucleus and in the Kinetoplast1, 2

SYNOPSIS. Autoradiographic technics with tritium-labeled thymidine have been used to determine G1, S, G2 and D for the kinetoplast and the nucleus of Crithidia fasciculata at 15, 25 and 32 C. The kinetoplast completes division before the nucleus at all 3 temperatures. The S phases of both organelles occur in approximate synchrony and are approximately equal in length but the nucleus begins and completes S before the kinetoplast at the 2 lower temperatures. This relationship is reversed at 32 C. Most of the effect of temperature on generation time is due to its effect on the length of S. The results are compared with similar studies on C. luciliae, Trypanosoma mega, other protozoa and tissue cells in culture. The role of the approximate synchrony of nuclear and kinetoplastic cycles in maintenance of the kinetoplastic condition is discussed and the hypothesis is proposed that this synchrony results from the sharing by nucleus and kinetoplast of the same mechanism for the production of the deoxyribonucleotides used in replication of their respective DNAs.     

Evolutionary Ethics: An Irresistible Temptation: Some Reflections on Paul Farber‘s The Temptation of Evolutionary Ethics

In his recent The Temptation of Evolutionary Ethics, Paul Farber has given a negative assessment of the last one hundred years of attempts in Anglo-American philosophy, beginning with Darwin, to develop an evolutionary ethics. Farber identifies some version of the naturalistic fallacy as one of the central sources for the failures of evolutionary ethics. For this reason, and others, Farber urges that though it has its attraction, evolutionary ethics is a temptation to be resisted. In this discussion I identify three major, historically relevant forms of the naturalistic fallacy, the (1) the deductive, (2) genetic, and (3) open question forms and argue that none of them pose an intrinsic problem for evolutionary ethics. I conclude that on this score at least there is no reason to resist temptation.     

Herpesviruses in Metastatic Lucké Renal Adenocarcinoma

The etiologic agent of the renal adenocarcinoma of leopard frogs, Rana pipiens , is the Lucké tumor herpesvirus (LTHV). The virus is easily detected with thin section electron microscopy in primary tumors of frogs which have been exposed to a cold environment. Several spontaneous metastatic nodules and a large primary tumor were detected at autopsy of a frog which had been maintained at 4°C for 73 days. LTHV was found not only in the primary tumor, as previously reported, but also was present in metastatic tumor cells in the liver, fat body, and bladder. The presence of LTHV in metastatic cells demonstrates that the differentiated state of primary Lucké tumor cells is retained in its metastatic colonies even at the fine structure level revealed by electron microscopy.     

Flagellate nutritional versatility as a key to survival in two contrasting Antarctic saline lakes

1. Seasonal patterns of grazing and photosynthesis were investigated in two saline Antarctic lakes (Highway and Ace) in the Vestfold Hills (68°S). The phototrophic nanoflagellate (PNAN) community was dominated by Pyramimonas gelidicola and two morphological forms of a cryptophyte species that occurred throughout the year. Both species were mixotrophic on bacteria, and in Highway Lake they also exploited dissolved organic carbon as determined by the uptake of fluorescently labelled dextrans. 2. Clearance rates ranged between 0.02 and 0.21 nL h^?1 cell^?1 in Ace Lake and 0.004–1.05 nL h^?1 cell^?1 in Highway Lake. On occasion cryptophyte grazing equalled that of the heterotrophic nanoflagellates (HNAN). 3. Photosynthetic rates showed similar trends in both lakes, but there were differences in chlorophyll a specific rates and photosynthetic efficiency, probably related to the meromictic characteristic of Ace Lake. Primary production was measurable in winter and peaked in summer following the maxima of mixotroph grazing. 4. The HNAN community of Highway Lake achieved clearance rates of 0.02–1.80 nL h^?1 cell^?1 and removing between 50 and 693 ng bacterial carbon L^?1 day^?1, with highest impact in winter when HNAN were most abundant. The HNAN also ingested fluorescently labelled dextrans showing a preference for 4 and 500 kDa molecules. The more diverse HNAN community of Ace Lake had lower clearance rates (0.04–0.37 nL h^?1 cell^?1) and exerted a lower grazing pressure on bacterioplankton. In Highway Lake, where the HNAN community was dominated by the choanoflagellate Diaphanoeca grandis, there was a significant correlation between mean cell volume and clearance rate. 5. The major feature was that the microbial plankton functioned throughout the year by employing nutritional versatility.     

Biomass and Pigment Production in Three Isolates of Azolla I. Response to Water Stress

Two isolates of Azolla Lam. from Colombia, South America, andone from the USA (Florida) were selected for characterizationof their growth potentials in certain defined stress environments.These isolates were Amazonian (RAR) and Atlantic coastal (M-3)specimens of Azolla caroliniana Willd and a Floridian variety(LA) of Azolla filiculoides Lam. Osmotic stress (from polyethylene glycol-amended media) andosmotic/salt stress (from seawater-amended media) were the chosendeterminants in these trials. Their effects on growth responseswere monitored. Initial growth rates of LA and M-3 were stimulatedat 1 and 5 ppt seawater salts, relative to growth under controlconditions. RAR, however, was generally the most stable isolatein the experiments with seawater-induced ionic stress. LA wasmost stable in its biomass growth in experiments with polyethyleneglycol 6000, but chlorophyll accumulation data were again leastaffected in RAR. LA survived the longest of the three isolatesat a water potential of approximately —10 bars with eitherseawater (13 ppt salts) or polyethylene glycol (26 per cent).The latter amendment was usually more inhibitory to acetylenereduction rates in RAR and consistently more inhibitory in LA.RAR was tentatively judged by its reactions to water stressto be the superior isolate of the three. Azolla caroliniana Willd., Azolla filiculoides Lam., biomass and chlorophyll accumulation, acetylene reduction, osmotic/salt stress     

Metamorphosis from the Phoront to the Trophont in Hyalophysa*

SYNOPSIS. The nature and sequence of changes involved in the metamorphosis of the phoront of Hyalophysa have been examined in the light microscope using both living and silver-impregnated ciliates. The phoront's infraciliature, which resembles that of the migratory tomite, differentiates to the trophont (feeding stage) infraciliature without an intervening dedifferentiation. The primary visible event of metamorphosis is the growth of the metastomial area that distorts the meridional somatic kineties so that they curve or spiral around it. The distinct borders and intense argentophilia of the metastomial area suggest that it is a discrete organelle which is probably involved in the management and concentration of a large volume of dilute food. The anterior ventral field of kinetosomes, whose presence distinguishes Hyalophysa from Gymnodinioides, forms from a disorganized anterior segment of falciform field 9. The rapid and pronounced elongation of the somatic kineties supports our view that the kinetodesmos elongates by the sliding of its component subfibrils upon one another, and that the accessory kinetosomes seen in the electron microscope become functional at metamorphosis. The movement of the kineties and contractile vacuole pore during metamorphosis suggests that differentiation in this instance is not directed by pellicular “fields” but by morphogenetic movements of the underlying cytoplasm.     

The Effect of Cadmium on Net Photosynthesis, Transpiration, and Dark Respiration of Excised Silver Maple Leaves

Excised leaves of silver maple (Acer saccharinum L.) exposed to 0, 0.045, 0.090, or 0.180 mM Cd²⁴ exhibited reduced net photosynthesis and transpiration, and increased dark respiration. Rates of net photosynthesis and transpiration diminished with time and were strongly correlated with solution concentration and tissue content of Cd²⁴, Net photosynthesis and transpiration were reduced to 18 and 21%, respectively, of the untreated controls after 64 h. Dark respiration increased as much as 193% of the untreated controls but was poorly correlated with solution concentration or tissue content of Cd²⁴, Diffusive resistances of leaves to carbon dioxide and water vapor transfer increased with both increasing Cd²⁴ concentration and time. These findings are discussed in relation to stomatal function.     

Changes in dissolved organic material determine exposure of stream benthic communities to UV-B radiation and heavy metals: implications for climate change

Changes in regional climate in the Rocky Mountains over the next 100 years are expected to have significant effects on biogeochemical cycles and hydrological processes. In particular, decreased discharge and lower stream depth during summer when ultraviolet radiation (UVR) is the highest combined with greater photo-oxidation of dissolved organic materials (DOM) will significantly increase exposure of benthic communities to UVR. Communities in many Rocky Mountain streams are simultaneously exposed to elevated metals from abandoned mines, the toxicity and bioavailability of which are also determined by DOM. We integrated field surveys of 19 streams (21 sites) along a gradient of metal contamination with microcosm and field experiments conducted in Colorado, USA, and New Zealand to investigate the influence of DOM on bioavailability of heavy metals and exposure of benthic communities to UVR. Spatial and seasonal variation in DOM were closely related to stream discharge and significantly influenced heavy metal uptake in benthic organisms. Qualitative and quantitative changes in DOM resulting from exposure to sunlight increased UV-B (290–320 nm) penetration and toxicity of heavy metals. Results of microcosm experiments showed that benthic communities from a metal-polluted stream were tolerant of metals, but were more sensitive to UV-B than communities from a reference stream. We speculate that the greater sensitivity of these communities to UV-B resulted from costs associated with metal tolerance. Exclusion of UVR from 12 separate Colorado streams and from outdoor stream microcosms in New Zealand increased the abundance of benthic organisms (mayflies, stoneflies, and caddisflies) by 18% and 54%, respectively. Our findings demonstrate the importance of considering changes in regional climate and UV-B exposure when assessing the effects of local anthropogenic stressors.