Reproductive aspects of American minks (Neovison vison) in central Spain: Testing the effects of prey availability

Some reproductive components of fitness (breeding times, litter size and the proportion of breeding females per year) of two feral populations of American minks in central Spain were investigated by means of direct observation from 2006 to 2010. The effects of prey availability were explored. Mating season was from February to March. Parturition dates did not differ between the two sites and pooled births took place between April (33.3%) and July (2.8%), peaking in May (40.3%). Mean litter size (the number of small cubs following their mother) was 3.43 ± 1.01 cubs (±SD; n = 30) with statistical significant differences between the two study sites. The strong seasonality in births and differences in litter size were related to prey availability (i.e. these are food-limited life-history traits), but in a complex non-linear fashion. A minimum value of prey abundance is necessary to breed. While litter size rises with prey abundance, there is a point where increasing prey did not result in an increase in litter size. Up to 75% of the females in the breeding pool reared cubs each year in both areas, which represents a somewhat high pool of breeding females. Delayed implantation is shorter than a month or does not exist in these populations. Such a short time-span seems to emerge because longer delays would have an important fitness cost to females. The breeding calendar and the litter size were similar to that reported previously from other areas.     

Time-lapse videography of human oocytes following intracytoplasmic sperm injection: Events up to the first cleavage division

A total of 341 fertilized and 37 unfertilized oocytes from 63 intracytoplasmic sperm injection (ICSI) treatment cycles were included for retrospective assessment using the Embryoscope™ time-lapse video system. The second polar body (pb2) extrusion occurred at 2.9 ± 0.1 h (range 0.70–10.15 h) relative to sperm injection. All oocytes reduced in size following sperm injection (p < 0.05) with shrinkage ceasing after 2 h in the unfertilized and at pb2 extrusion in the fertilized oocytes. Pb2 extrusion was significantly delayed for women aged >38 years compared to those <35 years (3.4 ± 0.2 vs. 2.8 ± 0.1, p < 0.01) or 35–38 years (3.4 ± 0.2 vs. 2.8 ± 0.1, p < 0.01), but timing was not related to the Day 3 morphological grades (1–4) of subsequent embryos (2.9 ± 0.1, 2.9 ± 0.1, 2.8 ± 0.2 and 3.0 ± 0.1; p > 0.05 respectively). A shorter time of first cleavage division relative to either sperm injection or pb2 extrusion is associated with both top grade (AUC = 0.596 or 0.601, p = 0.006 or 0.004) and usable embryos (AUC = 0.638 or 0.632, p = 0.000 respectively) on Day 3. In summary, (i) pb2 of human oocytes extrudes at various times following sperm injection, (ii) the timing of pb2 extrusion is significantly delayed when female age >38 years, but not related to subsequent embryo development, (iii) all human oocytes reduce in size following sperm injection, (iv) completion of pb2 extrusion in the fertilized oocytes is a pivotal event in terminating shrinkage of the vitellus, and (v) time to first cleavage division either from sperm injection or pb2 extrusion is a significant predictive marker for embryo quality on Day 3.     

Line and birth season effects on plasma testosterone and oxidative stress parameters in testis of maturing rabbits

Oxidative stress plays a key role in the male reproductive function. Differences between rabbit breeds have been found for testis size, seminiferous tubule diameter, number and size of interstitial and germ cells, etc. Traits related to the redox system could also be affected by genetic factors. It is likely that differences between breeds for these traits would lead to differences in reproductive maturation and fertility.We have investigated in the present paper the age-related changes of the plasma testosterone (TST) concentration, superoxide anion (O₂^?) radical formation, superoxide dismutase (SOD) activity, catalase (CAT) activity and thiobarbituric acid-reactive substances (TBARs) level in testis of rabbits in two breeds selected for different aptitudes (Caldes for growth rate and Prat for litter size). The effect of birth season for these traits was also assessed.Major changes in parameters related to oxidative stress were observed at an early age and most probably can be explained by the concomitant changes in testicular structure and function. Both lines showed similar developmental profiles and levels for all the variables studied. There was no interaction between line and birth season, consequently environmental conditions affected both lines in the same manner. Significant differences between males born in different seasons were found for O₂^? (4.84 ± 0.19 RLU/mg tissue min versus 5.67 ± 0.19 RLU/mg tissue min), SOD (6.12 ± 0.11 U/mg protein versus 7.09 ± 0.11 U/mg protein) and CAT (0.058 ± 0.002 K/mg protein versus 0.040 ± 0.002 K/mg protein). Future studies should take into account differences between seasons for a more precise analysis.     

The influence of maternal thermal environments on reproductive traits and hatchling traits in a Lacertid lizard,Takydromus septentrionalis

The thermal environment can induce substantial variation in important life-history traits. Experimental manipulation of the thermal environment can help researchers determine the contribution of this factor to phenotypic variation in life-history traits. During the reproductive season, we kept female northern grass lizards, Takydromus septentrionalis (Lacertidae), in three temperature-controlled rooms (25, 28 and 32 °C) to measure the effect of the maternal thermal environment on reproductive traits. Maternal thermal environment remarkably affected reproductive frequency and thereby seasonal reproductive output, but had little effect on reproductive traits per clutch or hatchling traits. Females kept at 32 °C produced more clutches and thus had shorter clutch intervals than females from 28 to 25 °C. Clutch size, clutch mass, relative clutch mass, egg size and hatchling traits did not vary among the three treatments. The eggs produced by the females were incubated at 27 °C and the traits of hatchlings were measured. The result that egg (offspring) size was independent of maternal thermal environments is consistent with the prediction of the optimal egg size (offspring) theory. The eggs produced by low temperature females (28 and 25 °C) took longer time to complete their post-oviposition development than did eggs produced by high temperature females (32 °C). This suggests that the eggs from low temperatures might have been laid when the embryos were at relatively early stages. Therefore, maternal thermal environment prior to oviposition could affect post-oviposition development in T. septentrionalis.     

Ultrasonographic imaging of the reproductive tract and surgical recovery of oocytes in Cebus apella (capuchin monkeys)

Two-dimensional real-time and Doppler ultrasonography are valuable non-invasive methods to assess reproductive anatomy and physiology. In adult, postpubertal female Cebus apella (capuchin monkeys), the objectives were to determine (1) uterine and ovarian dimensions, ovarian follicular dynamics, day of ovulation, and arterial blood flow of uterus and utero-ovarian ligament during the follicular phase of the menstrual cycle and (2) the number of oocytes aspirated from antral follicles at laparotomy. Based on two-dimensional, transabdominal B-mode ultrasonography, mean (± S.E.M.) length, height, width, and volume of the uterus were 17.9 ± 0.4, 12.4 ± 0.3, 13.6 ± 0.3 mm, and 1.55 ± 0.08 mL, respectively, and of the ovary were 13.4 ± 0.2, 8.2 ± 0.1, 7.7 ± 0.1 mm, and 4.5 ± 0.2 mL. Ovarian follicles were monitored for 6 days before ovulation, which occurred on day 9.3 ± 0.5 (range, days 7–11; day 1 = start of menses), with 10 of 12 ovulations in the right ovary. Diameter and volume of the preovulatory follicle were 10.1 ± 0.2 mm and 0.55 ± 0.03 mL (on the estimated day of ovulation) and of the CL were 8.1 ± 0.4 mm and 0.3 ± 0.05 mL. Resistivity and pulsatility indices were 0.86 ± 0.02 and 2.15 ± 0.11 for uterine arteries, and were 0.69 ± 0.04 and 1.63 ± 0.15 for the utero-ovarian ligament (UOL) artery; just prior to ovulation, both indices peaked (P < 0.05) in the uterine artery ipsilateral to the side of ovulation, but both reached a nadir (P < 0.05) in the UOL artery. In the absence of ovarian stimulation, 31 oocytes (diameter, 137 ± 10 μm) were aspirated (average of 2 oocytes/(female attempt)) on days 5, 7, and 9. In conclusion, transabdominal ultrasonography facilitated assessment of reproductive anatomy and physiology in C. apella adult females. Resistance and pulsatility indices of uterine and UOL arteries changed near the time of ovulation. Dominant follicles were easiest to aspirate at 8–9 mm in diameter (∼day 9), with intact cumulus-oocyte complexes recovered from ovarian follicles 2–9 mm in diameter.     

Assessment of the reproductive parameters,laparoscopic oocyte recovery and the first embryos produced in vitro from endangered Canindé goats (Capra hircus)

The Canindé breed of goats (Capra hircus) is currently endangered. The aims of this study were to characterize the estrus behavior, ovulatory responses and progesterone profiles, and to evaluate the in vitro embryo production (IVP) in this breed. In Experiment 1, ten nulliparous and seven pluriparous females received medroxyprogesterone acetate (MAP)-containing sponges (60 mg) plus 75 μg d-cloprostenol for estrus synchronization and their reproductive parameters were evaluated. In Experiment 2, oocytes obtained by laparascopy from hormonally stimulated females (n = 15) were used for IVP. There was no difference (p > 0.05) between nulliparous and pluriparous goats in terms of estrus response (40.0% vs. 85.7%), time from progestagen sponge removal to the onset of estrus (62.0 ± 15.5 vs. 50.7 ± 19.2 h; mean ± SEM), duration of estrus (25.0 ± 16.1 vs. 30.0 ± 15.1 h), percentage of ovulating animals (60.0% vs. 85.7%), number of ovulations (1.2 ± 0.4 vs. 1.3 ± 0.8), and diameter of the preovulatory follicle (5.8 ± 0.5 vs. 6.1 ± 0.3 mm). Progesterone concentrations were also similar (p > 0.05) in both groups. During laparoscopic recovery, there were average 12.2 aspirated follicles and 9.1 oocytes per goat, resulting in a high recovery rate (74.3%, 182/245). A total of 78 embryos were produced (51.0%). The mean number of cells in the blastocysts at day 7 of in vitro culture was 170.3 ± 12.5. In conclusion, nulliparous and pluriparous Canindé goats exhibited similar reproductive profiles. It was possible to produce embryos in vitro, allowing the instigation of an embryo bank for preservation of this breed.     

Differences in preovulatory follicle dynamics and timing of preovulatory LH surge affect fertility of maiden sheep reared in semi-arid extensive conditions

In the current study follicular dynamics, pituitary function, ovulatory response and luteal activity of 30 maiden Barbarine sheep were analyzed according to oestrus occurrence and lambing outcome after oestrus synchronisation with cloprostenol. Animals were retrospectively classified in three groups named as O? (n = 7, ewes not displaying oestrus), O+L? (n = 7, ewes showing oestrus but failing to lamb) and O+L+ (n = 16; ewes showing oestrus and lambing thereafter). All the sheep ovulated and daily transrectal ultrasonographies revealed that preovulatory follicles were present at cloprostenol injection in all the animals. In sheep O+L+ and O+L?, 50% and 57% of the ovulatory follicles were the largest follicles at cloprostenol treatment (mean size of 4.1 ± 0.26 mm and 4.3 ± 0.74 mm, respectively). In O? ewes, the same percentage was higher (86%, P < 0.05 when compared to group O+L+; mean size of 4.0 ± 0.46 mm). The number of large follicles and the final diameter of the ovulatory follicles at oestrous tended thereafter to be higher in group O+L+ (1.4 ± 0.1 and 6.4 ± 0.2) than in groups O+L? (1 ± 0.2 and 5.7 ± 0.36) and O? (0.9 ± 0.2 and 5.9 ± 0.5, respectively). Conversely, the number of medium follicles at oestrus detection was higher in the group O+L? (2.1 ± 0.3, P < 0.05) than in the other two groups (1 ± 0.2 and 1 ± 0.3 for O+L+ and O? respectively). Timing of preovulatory LH surge was earlier for ewes O? (24.0 ± 4.75, P < 0.05) than for sheep O+L+ and O+L? (37.9 ± 2.45 h and 38.0 ± 4.75 h, respectively) and 94% of O+L+ ewes had a LH surge between 16 h and 64 h after cloprostenol injection compared to 57% in O+L? and O? groups (P < 0.05). Thus, maiden Barbarine sheep failing to display oestrus or conceive showed alterations in their follicular dynamics and, thereafter, pituitary function and ovulatory response.     

Trophic niche overlap and wild ungulate consumption by red fox and wolf in a mountain area in Italy

Red fox (Vulpes vulpes) and wolf (Canis lupus) are two widespread opportunistic predators living in simpatry in many areas. Nonetheless, scarce information are available on their trophic interactions. We investigated food habits of these two carnivores in a mountain area in Italy and assessed the extent of their trophic niche overlap, focusing on the consumption of wild ungulates. Thereby we analyzed the content of 669 red fox scats and 253 wolf scats collected between May 2008 and April 2009. Red foxes resulted to have a more than three times higher niche breadth than wolves. Vegetables, small mammals, wild ungulates, and invertebrates were major items (altogether 92% of volume) of the red fox annual diet. On the contrary wolf annual diet relied on wild ungulates (94% of volume) with wild boar (Sus scrofa) being the main food item. The degree of trophic niche overlap between the two species was found to be low (Pianka's O = 0.356). Diet variation between the warm and the cold seasons was limited in both species, and higher in red fox than in wolf. The two canids appeared to use wild ungulates unevenly being the former more selective for younger preys, smaller in size (newborn piglets and roe deer Capreolus capreolus fawns), whereas the latter exhibited a preference for medium-sized and large ungulates (10–35 kg wild boar and adult roe deer). Even if wild ungulates represent the main shared food category, the different use of age/weight classes by the two predators, together with their possible consumption as carrions by red fox, suggests a very limited trophic competition between wolf and red fox.This study represents a contribution to the knowledge of trophic interaction in predator–prey systems where sympatric carnivores are present.     

Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro

In vitro fertilization (IVF) and embryonic development of mature and meiotically arrested porcine oocytes were compared in the present study. After in vitro maturation (IVM) of cumulus-oocyte complexes for 48 h, 75.4% of them extruded a visible polar body (PB). Most of the oocytes with a first polar body (PB+ group) were at the metaphase-II (M-II) stage (91.4%). Most of the oocytes without a visible polar body (PB− group) appeared to be arrested at the germinal vesicle (GV) (41.6%) and metaphase-I (M-I) (34.0%) stages. After IVF of oocytes (day of IVF = Day 0), there was no difference between PB⁺ and PB⁻ groups in rates of sperm penetration, mono-spermy, however oocyte activation rate after penetration was greater in the PB+ than in the PB− group (P < 0.05). On Day 2, there was no difference between rates of embryos cleaved at the 2–4 cell stages in PB+ and PB− groups (42.1 ± 48.8% and 33.6 ± 2.1%, respectively). On Day 4, the rate of PB+ embryos developing beyond the 4-cell stage was greater than that of PB− embryos (P < 0.05, 31.7 ± 3.9% and 14.1 ± 1.5%, respectively), and PB+ embryos had more cells than the PB− embryos (P < 0.05, 8.3 ± 0.4 and 6.0 ± 0.8 cells, respectively). On Day 6, a greater proportion of PB+ embryos developed to the blastocyst stage than did PB− embryos (P < 0.05, 34.6 ± 2.4% and 20.7 ± 2.8%, respectively). However, when the GV oocytes of the PB− group were not included in recalculations, there was no difference in blastocyst rates between M-I arrested and M-II oocytes (35.3 and 34.6%, respectively). The number of blastomere nuclei in embryos obtained from the PB+ group (52.0 ± 2.5) was greater than that from the PB− group (P < 0.05, 29.1 ± 2.8). The proportion of degenerated parts in the blastocysts, as determined by morphological appearance, was the same in the PB+ and PB− groups. Although the quality of PB+ embryos was enhanced as compared with that of the PB− group, the proportion of inner cell mass and trophectoderm cells in PB+ and PB− blastocysts did not differ (1:1.9 and 1:2.2, respectively). Chromosome analysis revealed that PB+ blastocysts had more diploidy (P < 0.05, 69.7%) than did PB− blastocysts (44.0%), whereas PB− blastocysts had more triploid cells (P < 0.05, 34.0%) than did PB+ oocytes (8.4%). These results indicate that pig oocytes arrested before the M-II stage (M-I oocytes) undergo cytoplasmic maturation during maturation culture and have the same ability to develop to blastocysts after IVF as M-II oocytes, but some of them resulted in degeneration or delayed development with poor embryo quality.     

Biology and demography of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) on mulberry

Lesser mulberry pyralid, Glyphodes pyloalis Walker, is a monophagous pest of mulberry and has recently been reported in northern Iran. The biology and life table of this pest were studied in controlled conditions (24 ± 1 °C, 75 ± 5% RH and 16:8 LD). Mortality rate, metamorphosis, appearance of adult insects and adult sex ratio were recorded daily. Data were analyzed based on an age-stage, two-sex life table. Developmental rate among individuals and between sexes were also considered. The developmental periods for the egg, first through fifth instar larvae, prepupae and pupae were 4.06 ± 0.03, 2.93 ± 0.03, 2.03 ± 0.02, 2.01 ± 0.01, 2.10 ± 0.03, 4.09 ± 0.03, 2.04 ± 0.02 and 9.7 ± 0.09 days, respectively. The mean total developmental period from egg to adult was 35.40 ± 0.37 days. The maximum adult longevity was 7 and 11 days for males and females, respectively. The pre-oviposition period lasted 2.14 ± 0.04 days. Laboratory observation showed that adults are nocturnal and female lay eggs at night. The intrinsic rate of increase (r_m) and the finite rate of increase (λ) were 0.14 ± 0.005 d^? 1 and 1.15 ± 0.01 d^? 1. The net reproductive rate (R₀), gross reproductive rate (GRR) and mean generation time (T) were 134.67 ± 20.6 female offspring, 294.71 ± 4.07 eggs/female and 34.44 ± 0.2 days, respectively. Life expectancy of freshly laid egg was 32.15 days.     

Utilization of poultry litter pellets in meat goat diets

Forty-eight crossbred meat goats were used to determine if poultry litter pellets could be used as a protein source in the diets of growing meat goats. Goats were fed one of three 19–21% CP diets containing 0 (CON; n = 18), 20% (20PL; n = 12) or 40% poultry litter pellets (40PL; n = 18). In Experiment 1, 38 animals (n = 13 CON; n = 12 20PL; n = 13 40PL) were used. Goats were allowed a 23-day adjustment period and body weight (BW) and feed intake were measured every 7 days for 42 days. In Experiment 2, 10 males fed CON or 40PL (n = 5 per diet) were used in two metabolism trials at 93.7 ± 0.9 (Trial 1) and 121.7 ± 0.9 d of age (Trial 2). Goats were placed in metabolism crates and after a 3-day adjustment period, feed intake and fecal and urine output were measured and sampled daily for 7 days to determine diet digestibility. In Experiment 1, ADG (79 ± 8 g) and feed efficiency (130 ± 12 g per kg) were not influenced by diet. In Experiment 2, for both trials, organic matter and CP digestibility were similar between diets (80 ± 1 and 70 ± 3% for Trial 1, respectively and 63 ± 2 and 75 ± 7% for Trial 2, respectively). Dry matter digestibility was greater (P < 0.05) for CON (81 ± 1 and 82 ± 1% for Trials 1 and 2, respectively) when compared to 40PL (77 ± 1 and 75 ± 1% for Trials 1 and 2, respectively). The ADF (41 ± 4% for CON and 67 ± 4% for 40PL) and NDF (48 ± 4% for CON and 71 ± 4% for 40PL) were greater (P < 0.01) for 40PL compared to CON diet in Trial 1 only. Digestibility (GE) was higher (P < 0.05) for 40PL (83 ± 0.3%) compared to CON (82 ± 0.3%) in Trial 2 only. The poultry litter pellets may be used effectively as a short-term feedstuff for meat goats.     

Embryo production in superstimulated llamas pre-treated to inhibit follicular growth

Llamas are monotocous and the length of their gestation period varies between 342 and 350 days. Thus the average number of offspring any female can produce throughout her reproductive life is very limited to spread a desired genome. The multiple ovulation and embryo transfer (MOET) technique allows an alternative to this limitation and reduces the generation interval. The objective of this study was to evaluate embryo recovery in superstimulated llamas which had previously been hormone-treated to inhibit follicular growth. A total of 50 female llamas were monitored daily via rectal palpation and ultrasound and divided according to their ovarian follicular growth into four phases. The females in each phase were then randomly divided into two groups: A (n = 20) received a single dose of 1 mg of estradiol benzoate (EB) on the first day of the treatment + 100 mg of progesterone (P4) i.m. for 5 days with 5 animals per phase and B (n = 20) received 1 mg EB at onset + 150 mg P4 i.m. for a period of 5 days with 5 animals per phase. Group C (n = 10) or control did not receive any prior hormonal treatment and the females were in follicular phase I. All groups were monitored daily and, in the presence of ovarian follicles smaller than the dominant size at the end of treatment, all were superstimulated with 1000 IU eCG. For plasma progesterone concentration recording, daily blood samples were collected from days ?1 to 5 in the treated females in Group A and B. No significant differences were observed regarding the inhibition of follicle growth and in the plasma progesterone concentrations between Group A and B. The ovarian response to superstimulation was 56.2%, 71.4% and 90%, with the average number of dominant follicles produced per female being 4.4 ± 0.9; 4.8 ± 0.7 and 4.6 ± 0.6 in Groups A, B and C, respectively. The embryo recovery rate was 77.7%; 90% and 66.7% and the average number of embryos recovered per female was 2.9 ± 0.9; 2.6 ± 0.9 and 2.4 ± 0.8 for Groups A, B and C, respectively. In Groups A and B, the static follicular phase (III) seemed to be ideal for initiating the assisted reproductive technique of MOET. Although prior administration of P4 + EB seems to have no effect on the number of females that responded to the superstimulation treatments, the number of embryos recovered showed a tendency to be higher when ovarian follicle growth inhibition was performed beforehand.     

Experimental evaluation of the reliability of placental scar counts in American mink (Mustela vison)

Counting of placental scars in the uterus is a valuable method to estimate female fecundity in wild-ranging mammalian populations, but the reliability of placental scar counts (PSC) has only been thoroughly assessed for a few species. Here we evaluate the reliability of PSC in American mink (Mustela vison) through examination of farm animals with known reproductive history. PSC estimated pregnancy rate and litter size accurately in individuals up to 3 months postpartum. Placental scars were detectable in most female American mink sampled 7–8 months postpartum, but the proportion of minks misclassified as barren was overestimated, and mean litter size was underestimated. We found a large variation in the intensity of pigmentation in placental scars with similar age. Differentiation of scars according to shading as indicating either successful full-term pregnancies or post-implantation loss of embryos was not possible. We propose that these results are applicable for fecundity analysis in American mink and closely related mustelids.     

Body temperature,thermoregulatory behaviour and pelt characteristics of three colour morphs of springbok (Antidorcas marsupialis)

Using intra-abdominal miniature data loggers, we measured core body temperature in female springbok (Antidorcas marsupialis) of three colour morphs (black, normal and white), free-living in the Karoo, South Africa, for one year. During winter, white springbok displayed lower daily minimum body temperatures (37.4 ± 0.5 °C), than both black (38.1 ± 0.3 °C) and normal (38.0 ± 0.6 °C) springbok. During spring, black springbok displayed higher daily maximum body temperatures (40.7 ± 0.1 °C) than both white (40.2 ± 0.2 °C) and normal (40.2 ± 0.2 °C) springbok. These high maximum body temperatures were associated with larger daily amplitudes of nychthemeral rhythm of body temperature (2.0 ± 0.2 °C), than that of white (1.6 ± 0.1 °C) and normal (1.7 ± 0.2 °C) springbok. Biophysical properties of sample springbok pelts were consistent with these patterns, as the black springbok pelt showed lower reflectance in the visible spectral range, and higher heat load from simulated solar radiation, than did the pelts of the other two springbok. Black springbok had lower diurnal activity in winter, consistent with them having to forage less because their metabolic cost of homeothermy was lower, but were disadvantaged in hot periods. White springbok, by contrast, were more protected from solar heat load, but potentially less able to meet the energy cost of homeothermy in winter. Thus energy considerations may underlie the rarity of the springbok colour morphs.     

Hatching,survival and deformities of piracanjuba (Brycon orbignyanus) embryos subjected to different cooling protocols

Groups of one hundred Brycon orbignyanus embryos at the stage of blastopore closure were subjected to different cooling protocols. Different combinations and concentrations of cryoprotectants were tested: sucrose, methanol, ethylene glycol and dimethyl sulfoxide (Me₂SO); at different temperatures (0.0 ± 2.0 °C and 8.0 ± 2.0 °C) and refrigeration times (6, 10, 24, 72 and 168 h), with the exception of the positive control (incubation without previous cooling). At the end of each refrigeration time, the embryos were acclimatized, rehydrated and incubated to determine hatching, survival and deformity rates. Morphological analysis of embryos was also carried out. The results showed that temperature and refrigeration time are critical factors for embryo survival. No embryos survived after 24, 72 and 168 h of refrigeration. Furthermore, when the refrigeration time increased from 6 to 10 h and the temperature decreased from 8.0 ± 2.0 °C to 0.0 ± 2.0 °C, mortality rates increased significantly. It was also found that in all protocols dead eggs and/or larvae with some degree of deformity were present. The main larval deformities observed were the malformation of the head, tail, yolk sac, vertebral column and eyes.     

Effect of the timing of ice slurry ingestion for precooling on endurance exercise capacity in a warm environment

It has been demonstrated that precooling with ice slurry ingestion enhances endurance exercise capacity in the heat. However, no studies have yet evaluated the optimal timing of ice slurry ingestion for precooling. This study aimed to investigate the effects of varying the timing of ice slurry ingestion for precooling on endurance exercise capacity in a warm environment. Ten active male participants completed 3 experimental cycling trials to exhaustion at 55% peak power output (PPO) after 15 min of warm-up at 30% PPO at 30 °C and 80% relative humidity. Three experimental conditions were set: no ice slurry ingestion (CON), pre-warm-up ice slurry ingestion (−1 °C; 7.5 g kg⁻¹) (PRE), and post-warm-up ice slurry ingestion (POST). Rectal and mean skin temperatures at the beginning of exercise in the POST condition (37.1±0.2 °C, 33.8±0.9 °C, respectively) were lower than those in the CON (37.5±0.3 °C; P<0.001, 34.8±0.8 °C; P<0.01, respectively) and PRE (37.4±0.2 °C; P<0.01, 34.6±0.7 °C; P<0.01, respectively) conditions. These reductions increased heat storage capacity and resulted in improved exercise capacity in the POST condition (60.2±8.7 min) compared to that in the CON (52.0±11.9 min; effect size [ES]=0.78) and PRE (56.9±10.4 min; ES=0.34) conditions. Ice slurry ingestion after warm-up effectively reduced both rectal and skin temperatures and increased cycling time to exhaustion in a warm environment. Timing ice slurry ingestion to occur after warm-up may be effective for precooling in a warm environment.     

Population density and abundance of sympatric large carnivores in the lowland tropical evergreen forest of Indian Eastern Himalayas

Low density occurrence of large carnivore species and direct hunting of predators and prey make carnivore conservation complex. Vital baseline information on population status of large carnivores is still deficient in most forests of eastern Himalaya, which are known to be the biodiversity hotspots. To fill this information gap, we estimated the large carnivore population status and abundance in an intricate eastern Himalayan lowland tropical forest in Pakke Tiger Reserve, Arunachal Pradesh. Population status and abundance estimates of tigers and leopards were made through individual identification using closed capture-recapture sampling. To estimate the dhole abundance photographic encounter rate was used. For individually non-identifiable species photographic rate seemed to correlate well with animal abundance. The estimated tiger and leopard density through 1/2 MMDM was 2.14 ± 0.04/100 km² and 2.99 ± 1.13/100 km² respectively. Maximum likelihood estimates shows density of tiger 1.86 ± 0.7 and for leopard 2.82 ± 1.2.The estimated dhole abundance was (N) 10.6 ± 0.94, and density 6.62 ± 0.58 individuals in 100 km². Further, occupancy estimation of large carnivores may be tried along with assessing the comparative efficacy of other population estimation methods to establish better monitoring methods for this region.     

Genetic and phenotypic parameters for lactation traits in a flock of Saanen goats in Mexico

Heritability, repeatability and genetic and phenotypic correlations of total milk production (TM), cumulated milk production to 120 days (M120) and lactation length (LL), were estimated from the analysis of 1413 records (566 does) obtained from a Saanen goat flock in Querétaro, Mexico, during 1991–1997. Variance components were estimated using a restricted maximum likelihood procedure and applying an animal model. The model included the fixed effects of year–season of kidding, litter size and age of doe, and random effects of animal, permanent environment and residual. For TM, M120 and LL estimates of heritability were 0.22 ± 0.07, 0.12 ± 0.05 and 0.04 ± 0.03, and of repeatability were 0.40, 0.27 and 0.11, respectively. Genetic correlations between TM and M120, TM and LL, and M120 and LL, were 0.98, 0.58 and 0.53, respectively. Phenotypic correlations ranged from 0.30 to 0.83. The correlated selection response for TM from indirect selection on M120 was 71.5% of that expected from direct selection. Efficient selection for TM is possible and M120 could be used as an early indicator of TM, due to the high indirect selection response calculated.     

Influence of physiological concentrations of androgens on the developmental competence and sex ratio of in vitro produced bovine embryos

This study was designed to determine if the addition of androgens at ovarian follicular fluid (FF) concentrations to oocyte maturation media would alter the development and sex ratio of bovine embryos. To maximize hormone bioavailability, oil was removed and glass culture dishes were used during in vitro maturation (IVM) phase; this modified system was then used in the present experiment along with the standard IVM system utilizing plastic containers and incubation under oil. Ethanol (0.2%) was the vector for steroid hormone delivery. Oocytes were incubated for 22 h in the presence of two doses (“low” and “high”) of androstenedione (A₄) or testosterone (T); the doses were based on the concentrations of both androgens in preovulatory bovine follicles (A₄: 337.5 and 562.5 ng/ml; T: 22.2 and 42.6 ng/ml). The results of hormone assays indicated that bioavailability of steroid hormones remained relatively constant, regardless of the IVM system used. The plasticware with the addition of T resulted in significantly higher cleavage rates (80.0 ± 2.1%) than any other combination of treatments (plasticware × A₄: 71.5 ± 2.6%; glassware × T: 71.2 ± 1.9%; and glassware × A₄: 71.4 ± 2.4%). The blastocyst formation rate for the plasticware × T treatment (39.7 ± 2.5%) was significantly greater than for all other combinations (glassware × T: 28.7 ± 2.2%; glassware × A₄: 24.0 ± 2.8%; and plasticware × A₄: 19.8 ± 3.0%) and the low dose of T (37.1 ± 2.5%) resulted in higher (p < 0.05) blastocyst formation rates than all other treatments (T high dose: 29.2 ± 2.5%; A₄ high dose: 27.1 ± 2.9%; and A₄ low dose: 20.2 ± 3.0%). The proportion of male embryos was greater (p < 0.05) in plastic than glass dishes in the low-dose A₄ group (59.1 ± 8.7% vs. 38.2 ± 5.5%, plasticware vs. glassware, respectively) and it tended to be greater (p < 0.08) in the control groups and high-dose A₄ group, but not in the T groups. There was a moderate positive correlation between blastocyst formation rates across all treatment and control groups, and the percentage of male bovine embryos (r = 0.38, p < 0.05). In summary, specific combinations of androgen and glassware/plasticware treatments did alter early bovine embryo development and sex ratio. The addition of T to IVM media increased the cleavage and blastocyst formation rates in plasticware and may be employed to improve the efficiency of the standard in vitro embryo production systems. Androstenedione appeared to enhance whereas testosterone nullified the deviation in sex ratio (pro-femaleness) associated with the use of glass IVM dishes.