Dimensions and composition of eggs from captive bustards (Gruiformes: Otididae): houbara (Chlamydotis undulata), rufous‐crested (Eupodotis ruficrista), and kori (Ardeotis kori)

This study sought to increase understanding of the size and composition of eggs from two subspecies of houbara bustard (Chlamydotis undulata macqueenii and C. u. undulata). Eggs from the rufous‐crested bustard (Eupodotis ruficrista) and kori bustard (Ardeotis kori) were also examined for comparison. Infertile eggs were collected from captive birds; egg mass and linear dimensions were recorded, and egg component masses were determined wet and dry. Significant differences were observed in the composition of the eggs from the two houbara subspecies. Despite being smaller in size, eggs from the macqueenii subspecies had a relatively larger yolk (and relatively less albumen) than eggs from the undulata subspecies. The relative composition of the rufous‐crested and kori bustard eggs showed patterns similar to that of the undulata eggs. For the houbara bustards, changes in initial egg mass (IEM) were associated with changes in egg length more than egg breadth, and changes in egg length and breadth appeared to be due more to increases in albumen mass than to increases in yolk mass. Zoo Biol 21:337–346, 2002. © 2002 Wiley‐Liss, Inc.     

Proceedings of the New Zealand Society for Parasitology Inc.

Abstract

A study of the nesting habits and breeding biology of blue penguin Eudyptula minor was undertaken over the 1995–96 and 1996–97 breeding seasons on Matiu‐Somes Island, Wellington, New Zealand. Male and female blue penguins tended to be faithful to both mates and nest sites, although there was insufficient evidence to detect any association between a bird's breeding success in 1995 and a subsequent change of mate or nest in 1996. Over the 1995 and 1996 seasons the recorded hatching success (0.51 ±0.11 and 0.63 ± 0.10 respectively), fledging success (0.81 ±0.12 and 0.85 ±0.10 respectively) and reproductive success (0.41 + 0.11 and 0.54 ± 0.11 respectively) were similar each season. There was no significant difference between the proportion of eggs laid, or eggs hatched and chicks fledged, between the two seasons. The mean number of chicks raised over the two seasons was 0.94 ± 0.05 per nest. Replacement clutches were laid by 11 per cent of failed breeders in each season, but only in 1996 were they successful in fledging chicks.

No significant difference was found between the breeding success of the Matiu‐Somes Island blue penguin colony recorded during this study and a previous study undertaken on the island 40 years ago.     

Larviposition in the ovoviviparous blowfly Calliphora dubia

This study examined larviposition in Calliphora dubia Macquart (Diptera: Calliphoridae), an ovoviviparous blowfly of considerable forensic importance in Australia. Females in the field carried 22–83 live larvae, exhibiting a strong linear relationship between female size and the number of live larvae carried. Females took just over 1 min (mean 67.7 ± 7.7 s, n = 54) to larviposit live larvae on or near fresh liver in the laboratory. Females laid larvae at a mean rate of 1.2 ± 0.1 larvae/s, with the fastest rate being 3.4 larvae/s. Most females (70%) laid live larvae only, but 14% laid larvae and eggs at the same time and 16% laid eggs only (none of the eggs laid were viable). Females laying only live larvae laid a mean of 53.7 ± 2.3 larvae, whereas those laying only eggs laid a mean of 48.6 ± 2.8 eggs on each occasion. None of the eggs laid were viable. Most females (86%) laid all their larvae in a single spot, even if they engaged in several bouts of laying live larvae. Nearly one‐third of females did not lay all the live larvae in their ovisacs, but retained half of their complement of developed larvae. Females may be opting to spread their larvae across several carcasses in order to increase their survival and not to overcrowd small, ephemeral carcasses. The fact that a blowfly can lay either eggs or live larvae has enormous implications for the accurate determination of the post‐mortem interval (PMI) as the presence of larvae derived from eggs laid on the body add 6–18 h to the PMI. This paper represents the first report of the ability of female calliphorids to resorb some of their own live larvae.     

Effects of early rearing experience on adult behavior and nesting in captive Hawaiian crows (Corvus hawaiiensis)

Behavioral data were collected during the breeding season on eight pairs of Hawaiian crows (Corvus hawaiiensis) housed at two facilities on different islands. All data were collected from videocamera time‐lapse recordings of the nesting platforms. Behaviors included frequency of nest cup manipulation, percent of time spent on nest, allopreening, play, and stereotypy. The number of breeding pairs increased from four in 1996 to six in 1997, to seven in 1998, and to eight in 1999. Five of the older birds (three males and two females) were solitary‐reared for most if not all of their first year, while the remaining 12 birds were all socially reared. Significant differences were found between isolate‐ and socially‐reared birds, with isolate‐reared birds having higher rates of solo play (P = 0.0041) and stereotypies (P = 0.0090). Pairs that were comprised of at least one isolate‐reared bird engaged in significantly less allopreening (P = <0.0001) than pairs in which both birds were socially reared. From 1996 to 1999, 87 eggs were laid, with a mean of 1.88 ± 0.24 SEM eggs per clutch. Only three females produced clutches every year, and they were responsible for 85.1% of the eggs laid. Although not significant, the mean number of clutches produced per pair decreased from 2.50 ± 0.65 in 1996 to 0.87 ± 0.99 in 1999. Age of females does not appear to be a critical factor in the decrease in clutch production. New pairing combinations are under way in an effort to improve propagation in this highly endangered species. Zoo Biol 21:59–75, 2002. © 2002 Wiley‐Liss, Inc.     

Cues for communal egg‐laying in lizards (Bassiana duperreyi,Scincidae)

Animals may aggregate either because the presence of conspecifics provides information about habitat suitability, or because the presence of conspecifics directly enhances individual viability. For a female lizard, the advantage of laying her eggs in a communal nest may entail either information transfer (hatched eggshells show that the site has been successful in previous seasons) or direct physiological benefits (recently‐laid eggs can enhance water availability to other eggs). We tested the relative importance of these two mechanisms in the three‐lined alpine skink (Bassiana duperreyi Gray, 1838) by offering gravid females a choice between sites with hatched eggshells versus freshly‐laid eggs. Females selectively oviposited beside fresh eggs. In this species, early‐nesting females use information transfer (i.e. the presence of old eggshells) as a nest‐site criterion, whereas later nesters switch to a reliance on direct benefits of conspecific presence (i.e. the presence of freshly‐laid eggs). © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 839–842.     

Field and laboratory studies in a Neotropical population of the spinose ear tick, Otobius megnini

Abstract One ear of each of five cows on a property close to Dean Funes, province of Córdoba, Argentina, was inspected monthly from December 2004 to November 2006 to determine the presence of Otobius megnini (Dugès) and to ascertain its seasonality. Ticks were collected to study the biological parameters of larvae, nymphs and adult ticks. Groups of nymphs were also maintained at three different photoperiods at 25 °C. The abundance of immature stages was greatest during January–April and August–October in the first and second years of the study, respectively. No larvae successfully moulted. Nymphs weighing < 17 mg also failed to moult, but 89% of heavier nymphs moulted into adults. Nymphs moulting to males weighed less (49.5 ± 16.09 mg) than nymphs moulting to females (98.1 ± 34.08 mg). The pre‐moult period was similar for nymphs moulting to either sex and significantly longer (P < 0.01) for female nymphs maintained at 25 °C compared with nymphs kept at 27 °C. No effect of photoperiod on the pre‐moult periods of nymphs was detected. Female ticks produced a mean of 7.0 ± 1.94 egg batches after a preoviposition period of 16.4 ± 8.41 days for the first batch. The mean oviposition period was 61 ± 20.8 days and the duration of oviposition for each batch varied from 1 to 6 days. The mean number of eggs per batch was 93.1 ± 87.53. The minimum incubation period for the first egg batch was 13.6 ± 2.77 days. The total number of eggs laid by each female was 651.6 ± 288.90. Parthenogenesis was not observed. The reproductive efficiency index (REI) (number of eggs laid/weight of female in mg) was 5.5 ± 1.26. Pearson’s correlations showed a significant direct relationship between the weight of the female and number of eggs laid (P < 0.01) and REI (P < 0.05). Several of the biological values presented above for the tick population from the Neotropical zoogeographic region showed marked differences to equivalent values for O. megnini populations from the U.S.A. (Nearctic) and India (Oriental). Nevertheless, the only two sequences of 16S rDNA deposited in GenBank from ticks originating in Argentina and allegedly in the U.S.A. indicate that they are conspecific (99.8% agreement). We tentatively consider the biological differences among populations of this tick species to represent adaptations for survival at different conditions.     

Cryogenic preservation of semen from the Aleutian Canada goose (Branta canadensis leucopareia)

Aleutian Canada geese (Branta canadensis leucopareia) were inseminated with frozen-thawed semen containing 6% or 7% dimethylsulfoxide (DMSO) resulting in 32 fertile eggs and 17 goslings; with 7% DMSO, 19 of 31 eggs were fertile. Beltsville Poultry Semen Extender (BPSE), adjusted to 270 ± 30 mOs and 7.5 ± 0.4 pH, was used to dilute semen samples and the DMSO before cryopreservation. About half of the live spermatozoa in the fresh semen (92.9 ± 2.5% live cells, laboratory studies; 87.3 ± 7.3%, insemination trials) survived the freeze-thaw process (46.7 ± 7.8%, laboratory; 33.3 ± 17.8%, insemination trials). Samples of frozen-thawed semen contained a greater percentage of bent spermatozoa (27.1 ± 8.4% of live cells) than fresh semen (14.4 ± 3.0% of live cells). Fecal- and urate-contaminated semen (a common problem when collecting goose semen) reduced the sperm motility score from 3.2 ± 0.6 to 2.7 ± 0.7 and number of live spermatozoa in frozen-thawed semen from 49 ± 9% to 24 ± 18%. Other variables examined that had less of an effect on semen quality included semen extenders, semen holding temperature, dilution and equilibration, relationship between hour of semen collection and level of semen contamination, and the relationship between season and sperm concentration.     

The breeding biology of the Namaqua Sandgrouse,Pterocles namaqua

The breeding biology of the Namaqua Sandgrouse, Pterocles namaqua, was studied and its nesting success determined through the observation of 278 nests over four consecutive breeding seasons at Droëgrond, Northern Cape Province, South Africa. The normal clutch of three eggs is laid over five days (±48-hour laying interval). The incomplete clutch is left unattended overnight, but is attended during the heat of the day by the female on days when an egg is laid and by the male on alternate days. After clutch completion, the pair share incubation, with the female relieving the male 151 min (±21 S.D.) after sunrise and the male relieving the female 105 min (±21 S.D.) before sunset. The incubation period is 21 days from clutch completion, and the three chicks normally hatch within 18 hours of one another. Nesting success ranged from 5.7% to 13.5% between seasons and averaged 8.2%. Predation, primarily by small mammals, was responsible for 96% of nest losses. Estimates of annual recruitment at Droëgrond ranged from minima of 3–10% to maxima of 6–20%, and are believed to be representative of a core area of the distribution of the Namaqua Sandgrouse in South Africa. These low estimates suggest that annual juvenile recruitment may be too low to maintain Namaqua Sandgrouse populations locally. Possible reasons for the sustained low level of breeding success are discussed.     

Reproductive biology and ecology of white‐winged trumpeters (Psophia leucoptera) and recommendations for the breeding of captive trumpeters

The reproductive biology and ecology of a wild population of white‐winged trumpeters (Psophia leucoptera) were studied in southeastern Peru from 1983 to 1987. Because little information is available about any of the trumpeter species and because trumpeters have proven difficult to breed in captivity, information relevant to breeding and management of captive trumpeters is reported in this paper. White‐winged trumpeters lived in territorial social groups that ranged in size from four to 13 individuals. A typical territorial group contained three adult males, two adult females, and several sexually immature offspring, but smaller temporary groups sometimes formed for the duration of the breeding season. Only the dominant female contributed eggs to the clutch, and all adult males in the group competed to obtain copulations with her. Eggs were laid in elevated nesting cavities and no nest was constructed. The average clutch size was three eggs and incubation was not begun until the final egg was laid. The dominant male and female shared most of the incubation duties, but subordinate males covered approximately 15% of the incubation shifts. Eggs hatched approximately 27 days after incubation was begun and chicks left the nesting cavity the day after they hatched. Chicks were completely dependent on older birds to feed them for their first 3 weeks and then gradually began to feed themselves more and more food. The subordinate adult males fed chicks the most food, the dominant male and female and older offspring fed chicks an intermediate amount, and the subordinate adult female fed chicks the least. Young chicks behaved aggressively toward each other but were separated by adults before they injured each other. If at least one chick from the clutch survived, trumpeters did not breed again until the beginning of the next breeding season the following year. Zoo Biol 19:65–84, 2000. © 2000 Wiley‐Liss, Inc.     

Observations on the female reproductive cycles of captive Asian yellow pond turtles (Mauremys mutica) with radiography and ultrasonography

The annual reproductive cycle of 27 female Mauremys mutica was observed by radiography and ultrasonography from April 2006 to August 2007. Radiography was used to monitor clutch size and ultrasonography was used to monitor changes in the ovarian follicles. The follicles started to enlarge in September and became preovulatory in January. The mean maximum follicle diameter of ovulation was 18.30±1.44 mm, and ovulation occurred from March through August. Eggs were laid between April and August. Turtles entered latent period in early August and the maximum follicular size was at a low of 13.22±2.36 mm in late September. The vitellogenesis of the next reproductive cycle began in October. The 24 adult females laid 56 clutches containing a total of 227 eggs. Average clutch size was 4.05 eggs (range 1–8) and there were 2.33 clutches (range 1–4) per female. Egg shell images were first observed on the sixth or seventh day after ovulation. The oviductal period averaged 6.9 weeks (range 2–16 weeks) on the first clutch, 3.4 weeks (range 2–8 weeks) on the second, and 2.75 weeks (range 2–6 weeks) for the third. Radiography and ultrasonography are non‐invasive and convenient methods to evaluate the reproductive cycle of female M. mutica. They should be applicable to other turtles and should greatly enhance knowledge of reproductive physiology. Zoo Biol 29:50–58, 2010. © 2009 Wiley‐Liss, Inc.     

Breeding success in a Houbara Bustard Chlamydotis [undulata] macqueenii population on the eastern fringe of the Jungar Basin, People's Republic of China

Nesting success and chick survival of a migratory population of Houbara Bustard Chlamydotis [undulata] macqueenii were studied during three consecutive years (1998–2000) in the Xinjiang province of north‐west China. A total of 45 nests was monitored and 85 broods comprising 227 chicks were captured, of which 82 chicks were radio‐tracked. Start of laying varied between 6 and 17 April between years but the laying mode fell consistently between 26 and 30 April. Mean clutch size was 4.0 (sd = 0.8) (range 2–6) for early clutches and 3.3 (sd = 1.1) for late clutches (range 2–5). The average nesting success was 0.588 (sd = 0.270) but great variations were observed between years –0.882 in 1998, 0.530 in 1999 and 0.351 in 2000. This was related to increased predation in 1999 and 2000, which is reflected by increased predator density (chiefly Corsac Fox Vulpes corsac and Long‐legged Buzzards Buteo rufinus). The overall hatchability, defined as the proportion of eggs hatched in successful nests was 0.839 sd = 0.238). The average brood size at hatching varied from 2.9 (sd = 0.8) to 3.3 (sd = 0.9) according to years, and no significant decrease in brood size was observed in the first 5 days post‐hatching. In 1999 and 2000 the brood size diminished sharply (14% and 27%, respectively) in prefledging chicks. A further severe decrease (37%) was observed in fledglings in 2000, probably due to predation by raptors. For the 3 years of the study, a successful female Houbara would bring on average 2.3 (sd = 0.9) chicks to fledging and would have lost 30.2% (sd = 14.9%) of its brood to adversity during the rearing process. The proportion of females that lost their entire brood was 0.181 in 1998, 0.708 in 1999 and 0.453 in 2000. For the 3 years of the study, only 55.3% (sd = 26.3%) of the females hatching eggs brought chicks to fledging. The overall chick production was 0.827 per breeding female per year and the probability of an egg laid producing a fledgling of 8 weeks old was 0.190.     

Relationship between female body size and demographic parameters inBactrocera Malaysian A (Diptera: Tephritidae)

The effect of body size, as measured by the head width, of the female Bactrocera sp. Malaysian A (kept separately in sexual pairs) on the demographic parameters was investigated in the laboratory under ambient conditions of 28–30°C, 78–85% RH and natural photoperiod. Body size was shown to influence significantly all the demographic parameters. The expectation of life of females at eclosion from pupae was respectively for head widths of 1.6, 1.8, 1.9, 2.0 and 2.1 mm: 76.2, 73.4, 73.8, 102.4 and 115.2 days. The mean number of eggs laid per female in its life time was respectively: 86.4±48.7, 181.8±56.1, 229.7±72.6, 364.3±69.4 and 477.5±109.3 which was significantly different from one another (F=3.73,P<0.05) especially the two smaller sizes from the two larger sizes. The regression line for total eggs laid (Y) against head width (X) was Y=785.2X−1208.7 (R²=0.35,P<0.001). The net reproductive rate (R₀) was respectively 15.8, 34.0, 43.5, 66.9 and 88.8 eggs, while the intrinsic rate of increase (r) was respectivley 0.0435, 0.0538, 0.0670, 0.0665 and 0.0711. The results confirm that for mass rearing purposes, larger females which produce more offspring are to be preferred.     

Captive breeding for reintroduction: influence of management practices and biological factors on survival of captive kaki (black stilt)

An important component of the restoration strategy for the critically endangered kaki or black stilt (Himantopus novaezelandiae) is captive breeding for release. Since 1981 1,879 eggs were collected from wild and captive pairs, with birds laying up to four clutches. Eggs were incubated artificially and most chicks reared by hand until released as juveniles (about 60 days) or sub‐adults (9–10 months). Because survival in captivity is a significant determinant of the number of birds available for release, we wished to identify sources of variation in mortality to assess potential impacts of management on productivity. Hatchability was 78% for captive‐laid eggs and 91% for wild‐laid eggs. Survival of hatched eggs was 82% by 10 months of age for both wild and captive birds. Most egg mortality occurred early in incubation and around hatching: the timing of mortality was unaffected by whether birds were captive or wild, hybrid or pure kaki, or when eggs were laid. Heavier hatchlings showed higher initial survival, as did chicks from wild parents. Hatchlings from fourth‐laid eggs showed lowest survival, even though hatchling mass tended to increase with hatch order. Survival of chicks subjected to major health interventions was 69% after 4 months. No differences in survival were found between different genders, hybrids and pure kaki, hand‐reared or parent‐reared birds, chicks hatching early or late in the season, different seasons, different‐sized groups of chicks, chicks reared in different brooders, juveniles kept in different aviaries, and chicks from subsequent clutches. Birds subjected to minor health interventions were equally likely to survive as healthy chicks (82%). Survival was high despite aggressive management (quadruple clutching and collecting late in the season). Differences between captive and wild birds suggest further improvements could be made to captive diet. Wide variation in hatchability between parent pairs substantiates the practice of breaking up poorly performing pairs. Zoo Biol 0:1–16, 2005. © 2005 Wiley‐Liss, Inc.     

Semen cryopreservation of yellow croaker <Emphasis Type="Italic">Larimichthys polyactis</Emphasis>

The effects of various extenders and cryoprotectants on movable spermatozoa ratio (MSR), spermatozoa velocity (SV) and duration of spermatozoa motility (DSM) of post-thawed spermatozoa were examined. The MSR, SV and DSM of post-thawed sperm in artificial seminal plasma (ASP) extender were higher than those in marine fish Ringer’s solution (MFRS) extender (P < 0.01) and was not significantly different from that of fresh sperm. No significant differences were observed in the motility parameters between fresh spermatozoa and frozen-thawed spermatozoa cryopreserved with ASP extender supplement 10% EG (ethylene glycol) cryoprotectant. Using the above method, yellow croaker semen was cryopreserved with extender ASP and 10% EG. As a result, at the spermatozoa/egg ratio of 100,000:1, the fertilization rate and hatching rate of the frozen-thawed spermatozoa cryopreserved for 1 week or 1 year in liquid nitrogen (45.7 ± 3.2% and 27.2 ± 5.0% or 37.5 ± 4.4% and 27.2 ± 5.0%) were similar to that of fresh spermatozoa (51.0 ± 3.1% and 36.7 ± 2.2%). There was a small alternation of shape in cryopreserved spermatozoa compared with fresh spermatozoa. In conclusion, the optimal conditions for yellow croaker semen cryopreservation are ASP extender supplement 10% EG cryoprotectant. This is the first report on semen cryopreservation of yellow croaker Larimichthys polyactis.     

两种寄生蜂的卵巢结构与卵成熟特点初步观察

对斜纹夜蛾的内寄生蜂双斑侧沟茧蜂MicroplitisbicoloratusChen和南美斑潜蝇的外寄生蜂潜蝇姬小蜂Diglyhusisaea(Walker)的卵巢结构进行了研究。双斑侧沟茧蜂的2个卵巢管底部膨大有大量的卵,通过凹陷的颈部与萼区相连接,在萼区里大量成熟待产的卵浸泡在萼液中。产卵量随产卵时间延长而降低,在日产卵实验中,日产卵量202(±114SE),从1000至1800每2h的产卵量分别占日产卵量的38,32,17和13%;在一生产卵量实验中,9d产卵期,每头雌峰一生平均产卵量为502(±78SE)粒,第1d产卵量占一生总产卵量的42%。潜蝇姬小蜂的每个卵巢由3条卵巢管组成,每条输卵管中仅有1粒成熟卵,整个卵巢中只有6粒成熟卵,每次产卵数不超过6粒。结果表明,M.bicoloratus是早熟卵(proovigenic)类昆虫;D.isaea是同步卵(synovigenic)类昆虫。     

Reproductive parameters in captive hand‐reared black‐bellied sandgrouse

Flock breeders of black‐bellied sandgrouse originated from wild‐laid eggs collected in west central Morocco in 2003 and 2004, were hatched, and hand‐reared in captivity in the framework of a reinforcement population program. Three to five pairs of different ages were housed in an aviary. Egg‐pulling procedure was used and eggs incubated artificially. Breeding parameters, hatchability, and posthatching mortality were recorded. The black‐bellied sandgrouse showed a seasonal breeding pattern with a laying period extending from 7 to12 weeks. The clutch frequency varied from 3 to 7 clutches per female per season. The mean clutch size was 2.66±0.47 eggs, and the mean interclutch interval was 10±2.7 days. The average total egg production was 12±5.83 eggs per female varying with age from 8 to18 eggs per female. Egg hatchability of incubated eggs increased with age and varied from 37.5 to 72.2%. Chick mortality occurred only in the first week after hatching, averaging 60.5%. The obtained results showed that black‐bellied sandgrouse can be successfully bred in captivity and opened the possibility of controlling, managing, and maximizing their production for the reinforcement of the local declining wild populations. Zoo Biol 27:269–281, 2008. © 2008 Wiley‐Liss, Inc.     

Cryopreservation of Atlantic salmon Salmo salar sperm: effects on sperm physiology

The objective of this study was to determine the effect of freezing on the function in Atlantic salmon Salmo salar spermatozoa. The semen was frozen in Cortland's medium + 1.3M dimethyl sulphoxide + 0.3M glucose + 2% bovine serum albumin (final concentration) in a ratio of 1:3 (semen:cryoprotectant) as the treatment (T) and fresh semen as the control (F). Straws of 0·5 ml of sperm suspension were frozen in 4 cm of N₂L. They were thawed in a thermoregulated bath (40° C). After thawing, the percentage of spermatozoa with fragmented DNA [transferase dUTP (deoxyuridine triphosphate) nick‐end labelling (TUNEL)], plasma membrane integrity (SYBR‐14/PI) and mitochondrial membrane potential (ΔΨMMit, JC‐1) were evaluated by flow cytometry and motility was evaluated by optical microscope under stroboscopic light. The fertilization rates of the control and treatment semen were tested at a sperm density of 1·5 × 10⁷ spermatozoa oocyte^?1, by observation of the first cleavages after 16 h incubation at 10° C. In the cryopreserved semen (T), the mean ± s.d . DNA fragmentation was 4·8 ± 2·5%; plasma membrane integrity 75·2 ± 6·3%; mitochondrial membrane potential 51·7 ± 3·6%; motility 58·5 ± 5·3%; curved line velocity (V_CL) 61·2 ± 17·4 µm s^?1; average‐path velocity (V_AP) 50·1 ± 17·3 µm s^?1; straight‐line velocity (V_SL) 59·1 ± 18·4 µm s^?1; fertilization rate 81·6 ± 1·9%. There were significant differences in the plasma membrane integrity, mitochondrial membrane potential, motility, fertilization rate, V_CL, V_AP and V_SL compared with the controls (P < 0·05). Also the mitochondrial membrane potential correlated with motility, fertilization rate, V_CL and V_SL (r = 0·75; r = 0·59; r = 0·77 and r = 0·79, respectively; P < 0·05); and the fertilization rate correlated with V_CL and V_SL (r = 0·59 and r = 0·55, respectively).     

Captive hybridization of two geographically isolated pygmy angelfish species,Centropyge fisheri and Centropyge resplendens

This study documents the rearing of two pygmy angelfish species, Centropyge fisheri and Centropyge resplendens, and the early life history and reproduction of their hybrid offspring. A C. fisheri female, collected from Hawaii, and a C. resplendens male, captive‐bred from parental stock collected from Ascension Island, were maintained at the hatchery facility for 7 months. Continuous spawning was achieved at a photoperiod cycle of 14L:10D and a water temperature of 26·5° C, range ±1° C. Over the 110 day period, the C. fisheri female spawned 102 times, 57% of which resulted in embryos (fertilized eggs). The mean ±s.d. fecundity per spawn was 730 ± 459 eggs (range 52–1967). Fertility (% eggs that developed into embryos) of all eggs that were preserved was 22·4 ± 25·6%. A total of 235 hybrid juveniles were raised through metamorphosis with an average larval survival of 16·4%. Eight F₁ hybrid juveniles isolated for further study began to display signs of reproductive behaviour c. 300 days post‐hatch (dph). Spawn resulting in non‐fertile eggs were first obtained 319 dph, and fertilized eggs developing into embryos were obtained after 411 dph from at least two female individuals. While no attempt was made at rearing the F₂ larvae, embryo and larval development were normal up to 8 dph. Reproduction and development observed for all hybrid generations in this study were normal, similar to other Centropyge species and indicates a very close phylogenetic relationship between what are currently considered distinct species, e.g. C. fisheri and C. resplendens.     

Fertilized eggs obtained from transplantation of frozen ovaries and parthenogenesis in combination with artificial insemination of frozen semen of the silkworm,Bombyx mori

A reliable method is reported for the long-term preservation of ovaries and spermatozoa of the silkworm (Bombyx mori). Three studies are presented. In the first, ovaries were removed from larvae at either 3rd, 4th, or 5th instar, cryopreserved, and stored in liquid nitrogen. Thawed ovaries were transplanted to surgically castrated female larvae at the same or a different developmental stage. The highest percentage of recipient females producing eggs resulted into either 3rd or 4th instar larvae (respectively, 22.1 and 8.7%). Similarly, the highest levels of other measurements of successful cryopreservation and transplanted ovary, and number of eggs laid, occurred with the same combination of donor and recipient developmental stages. Other combinations of ovary/recipient developmental stages yielded lower results. In the second experiment, semen was collected from male moths, cryopreserved, and then thawed semen was diluted with trypsin solution and artificially inseminated into females obtained from the best conditions of first experiment. A small percentage of inseminated moths laid eggs (8-10.3%) compared to that of controls (100%). In addition, the fertility of eggs from experimental moths was lower than that of control females (respectively, 40.3-88% and 97.5%). In the third experiment, eggs were surgically removed from ovarian tubules of moth following transplantation of thawed ovaries and subjected to parthenogenetic activation and artificial hatching. As expected, all resulting moths were female and, following natural mating or artificial insemination with thawed semen, yielded normal offspring at high rates.