352 Abstracts / Cryobiology 65 (2012) 339–366

with those grown in media with a 6:1 C:N ratio. OOT of cells and of lipidic extracts Conflict of interest: None declared.
were influenced by cellular carotenoids. E-mail address: alfredoramirez@uach.cl (A. Ramírez)
Fatty acid stability in frozen microalgae could be evaluated by analyzing the shift
of OOT values: the OOT values were lower for the frozen cells compared to the fresh
http://dx.doi.org/10.1016/j.cryobiol.2012.07.041
ones and decreased as a function of frozen storage time. This behaviour could be asso-
ciated with DHA degradation, in addition to carotenoid content lost.
DSC onset denaturation temperatures clearly correlated with protein stability and
the decrease in the enthalpy values obtained from the thermograms can provide valu-
41. Expansion of Ross’s gull nesting range in the north of Yakutia in the 20th century.
able information about the corresponding effect on yeast viability. Besides, lipid sta-
A.G. Degtaryov 1, K.S. Solomonov 2, N.G. Solomonov 3, N.I. Germogenov 3, 1 Department
bility in frozen media could be evaluated in whole microalgae cells as well as in their
of Biological Resources, Ministry of Nature Protection, SR(Y), Yakutsk, Russia,
lipidic extracts analyzing the shift of OOT values. DSC has a high potential, as a non- 2
Ammosov North-Eastern Federal University, Yakutsk, Russia, 3 Institute of Biological
chemical method, to determine stability parameters in complex biological systems
Problems of Cryolithozone SD RAS, Yakutsk, Russia
and could be employed as an efficient, reliable and fast method to study proteins
and fatty acids stability in whole cells and in their extracts.
This work was supported by CONICET (PIP 100846, 100468), UBA The nesting sites of Ross’s gull – the North’s legendary bird, first discovered on
(2002100100397 and I038) and ANPYCT (PICT 0928). the northern shores of Canada in 1823 and frequently encountered by Polar
Conflict of interest: None declared. researchers in different areas of the Arctic Ocean from the Canadian shores and
E-mail address: mag@di.fcen.uba.ar (M. Galvagno) East-Siberian and Chukchi Seas, were first discovered in 1905 by S.A. Buturlin in
the Kolyma River lower stream. Subsequent researchers identified the nesting
http://dx.doi.org/10.1016/j.cryobiol.2012.07.040 range of this beautiful bird from the Yana River delta to the Lower Kolyma and
Alazeya Rivers (Vorobyov, 1963). In the 1970–1980s the range of the species
expanded further to the west. Ross’s gull nesting in the Lena River delta was reg-
istered in 1982, 1983, 1984, 1988, 1990 and 1994. Local inhabitants noticed Ross’s
40. Development of stallion sperm cryopreservation extenders by analysis of semen quality gull even farther westward on the Olenyok River and its tributary Bur River
and new cryocapacitation markers. Alex Córdova 1, Pablo Strobel 1, Omar Ulloa 4, Javier (Labutin et al., 1985). Pavlov and Dorogov (1976) write about the Ross’s gull prop-
Urra 1, Carlos Contreras 1, Pamela Valenzuela 1, Alejandra Carrillo 1, Rafael Burgos 3, agation in Taimyr, in the Big Balakhnya River estuary. In early June 1994 we vis-
Bruno Menarim 2, Marcelo Ratto 1, Joan Enric Rodríguez-Gil 5, Alfredo Ramírez 1, ited Sagystyr Island located in the Big Tumatskaya Channel estuary on the
1
Instituto de Ciencia Animal, 2 Instituto de Ciencias Clínicas, 3 Instituto de Mor- northernmost tip of the Lena River delta, where we found three solitary nests
fofisiología y Farmacología, Facultad de Ciencias Veterinarias-Universidad Austral de of Ross’s gull. The first two nests were made on a tussock among small lakes. They
Chile, 4 Haras Militar Pupunahue, DGFER-Ejército de Chile, 5 Unitat Reproducció Ani- were made as cavities on the tussocks’ tops where soft moss was mixed with
mal, Universitat Autònoma de Barcelona thinly laid dry grass and dwarf willow leaves. The third nest was arranged on
the dry place near a geodesic tower and could be easily seen because of a white
gull sitting in a neighboring nest and well outlined against the brownish tundra.
Sperm capacitation is a complex process which is highly demanding of ATP con-
When the bird left the nest, the nest seemed to disappear as it melted into the
sidering events such as calcium flow, hyperactive motility, phosphorylation, and
surroundings becoming absolutely unnoticeable. Each nest contained three well
membrane-fusion, which are essential for gametes interaction and occurs in the
set eggs and the birds were sitting tightly. They flew up from the nest when a
female reproductive tract. Sperm cryocapacitation represents an altered state of
human came within 6–8 m, but they did not fly away and actively attacked the
sperm functionality triggered by cooling, freezing and thawing stress, so-called
stranger. Ross’s gulls winter in the areas of the Arctic Ocean, the Okhotsk and
due to similarities to the capacitation process, which happens precociously before
Bering Seas of the Pacific Ocean. In the last decades the Ross’s gull population
the insemination impeding fertilization. The aim of this study is to evaluate
has grown considerably in number. However, it still needs human protection. It
whether AMPK activation can stop or decrease the energetic processes linked to
is necessary to conduct permanent monitoring of the population of this amazing
sperm cryocapacitation, in order to increase the post-freezing quality in equine
endemic of the North and take all necessary protective measures.
semen using a freezing and experimental extender for equine semen considering
metabolic preconditioning as described by Menze et al.. (2010) in somatic cells.
Twelve ejaculates from Heavy Draft breeds stallions (3 Ardennes, 5 Percheron, 4 Conflict of interest: None declared.
Breton), were cryopreserved by standard procedures, using BotuCrioÒ (commercial Source of funding: None declared.
control) and experimental HM-0 extender as base medium to which Metformin, E-mail address: sks.inside@gmail.com (A.G. Degtaryov)
AICAR and AMP + compound C inhibitor were added. Post-centrifugation sperm pel-
lets were resuspended in extenders previously tempered at 20 °C, packed in 0.5 ml http://dx.doi.org/10.1016/j.cryobiol.2012.07.042
(50  106 sperm/mL) straws and cooled to 5 °C in 90 min. The straws were further
exposed to liquid nitrogen steam for 20 min, plunged in liquid nitrogen and stored
for at least 2 weeks at 196 °C. Samples were thawed at 37 °C for 30 s and objective
assessment of motility and viability were performed using the Sperm Class AnalyzerÒ 42. Age-old changes in ecosystems of Tit–Ary, the polar island in the lower reaches of the
System. Acrosomal integrity was evaluated using a conjugate of Pisum sativum agglu- Lena river (north-east Asia). A.P. Isaev 1,2, L.G. Mikhalyova 1, N.G. Solomonov 1,2, M.V.
tinin and fluorescein isothiocyanate (PSA-FITC) visualized by epifluorescence micros- Okoneshnikova 1, V.K. Vasilyeva 1, A.A. Popov 1, T.I. Ivanova 1, 1 Institute for Biological
copy and, simultaneous analysis of viability and acrosome integrity through flow Problems of Cryolithozone SB RAS, Yakutsk, Russia, 2 North-Eastern Federal Univer-
cytometry using propidium iodide and FITC-PSA. Phosphoproteins analysis were per- sity, Yakutsk, Russia
formed by Western blot and indirect immunofluorescence with the following anti-
bodies: p-AMPK, p-CaMKIV, P-DARPP-32, total phosphotyrosines (pY20) and b-
At the beginning of the 20th century, the expedition headed by Cajander (1903,
tubulin as loading control.
1904) discovered the northern outpost of a larch forest in the lower reaches of the
In vitro evaluation of frozen-thawed stallion semen indicates that of three indepen-
Lena River, on a polar island Tit-Ary. At that time, the canopy density of the forest
dently evaluated parameters (viability, acrosome integrity and motility), only motility
values reached 0.4; tree height was 3–5 m, and tree trunk stems were crooked. Two
was higher when BotuCrioÒ was used as the extender. However, motility did not differ
years later, the Australian ornithologists Hall and Trebilcock (Robin, Sirina, 2003)
between treatments when assessments were performed after one hour of incubation or
also explored the island, they discovered a number of migrant bird species includ-
in the presence of metabolic substrates such as glucose and/or fructose. Cytometric
ing sandpipers that spent winters in remote Australia.
analysis revealed a larger population of live sperms with intact acrosome using the
In 1942–1943, the forest was cut. In the 1950s, the course of restoration of the
experimental extender HM-0 (43.1 ± 3.3). However, no significant differences were
Tit–Ary forest was studied by Tikhomirov and Shtepa (1956). Later on, the island
found using HM-0 + AMPK activators such as Metformin (41.3 ± 3.6), AMP (41.1 ± 3.4)
was regularly surveyed by specialists from the Institute of Biology: in 1972, 1977,
and AICAR (39.3 ± 3.9), or with Compound C (37.3 ± 3.6), Compound C + AICAR
1982, 1984, 1989, 1994.
(32.7 ± 3.3) and BotuCrioÒ (34.8 ± 2.7). In addition, western blot analysis revealed sev-
During the last three years, a special investigation of the island ecosystems has
eral isoforms of p-AMPK and p-CaMKIV, for which AMPK did not respond to classic acti-
been conducted which revealed the following parameters of the forest: 2750 trees/
vators, but seemed that compound C inhibited its phosphorylation. Moreover, we also
ha, canopy density – 0.3, height – 1.5–3.2 (6.5) m, DBH – 1.7–6.2 (12) cm. After World
confirmed the presence of p-DARPP32 and its sensitivity to activation by NaHCO3/
War II, three basic waves of restoration were recorded in the 1960s, 1970s and 1980s.
cAMP, which makes it an early marker of the capacitation process.
The study of larch height increment proves that the climatic conditions in the begin-
The results of in vitro sperm evaluation validate the freezing protocol and the use
ning of the 21st century favoured the growth and development of larch at the north-
of HM-0, nonetheless, further studies improving our understanding of cellular mech-
ern limits of its distribution area. And, this is reflected in growth enhancement – the
anisms activated by cryopreservation stress, particularly related to sperm cryocapac-
average increment of axial shoot length during the last 10 years has increased from
itation will attempt to discover the importance/relevance of these and others
3.1 to 7.7 cm.
phosphoproteins in cryopreservation process.
Recent studies yielded an increase in the number of Tit–Ary vascular plant species
from 150 in the early 20th century to 206, moss species from 18 to 32, lichens from 5
Funding:FONDEF-CONICYT D08I1076 and DID-UACh S200961.