Spermatozoa from the maned wolf (Chrysocyon brachyurus) display typical canid hyper-sensitivity to osmotic and freezing-induced injury,but respond favorably to dimethyl sulfoxide

We assessed the influences of medium osmolality, cryoprotectant and cooling and warming rate on maned wolf (Chrysocyon brachyurus) spermatozoa. Ejaculates were exposed to Ham’s F10 medium (isotonic control) or to this medium plus NaCl (350–1000 mOsm), sucrose (369 and 479 mOsm), 1 M glycerol (1086 mOsm) or dimethyl sulfoxide (Me₂SO, 1151 mOsm) for 10 min. Each sample then was diluted back into Ham’s medium and assessed for sperm motility and plasma membrane integrity. Although glycerol and Me₂SO had no influence (P > 0.05), NaCl and sucrose solutions affected sperm motility (P < 0.05), but not membrane integrity. Motility of sperm exposed to <600 mOsm NaCl or sucrose was less (P < 0.05) than fresh ejaculate, but comparable (P > 0.05) to the control. As osmolality of the NaCl solution increased, motility decreased to <5%. In a separate study, ejaculates were diluted in Test Yolk Buffer containing 1 M glycerol or Me₂SO and cooled from 5 °C to −120 °C at −57.8 °C, −124.2 °C or −67.0 °C/min, frozen in LN₂, thawed in a water bath for 30 s at 37 °C or 10 s at 50 °C, and then assessed for motility, plasma- and acrosomal membrane integrity. Cryopreservation markedly (P < 0.05) reduced sperm motility by 70% compared to fresh samples. Higher (P < 0.05) post-thaw motility (20.0 ± 1.9% versus 13.5 ± 2.1%) and membrane integrity (51.2 ± 1.7% versus 41.5 ± 2.2%) were observed in samples cryopreserved in Me₂SO than in glycerol. Cooling rates influenced survival of sperm cryopreserved in glycerol with −57.8 °C/min being advantageous (P < 0.05). The findings demonstrate that although maned wolf spermatozoa are similar to domestic dog sperm in their sensitivity to osmotic-induced motility damage, the plasma membranes tolerate dehydration, and the cells respond favorably to Me₂SO as a cryoprotectant.     

The vocal repertoires of the bush dog (Speothos venaticus), crab-eating fox (Cerdocyon thous), and maned wolf (Chrysocyon brachyurus)

The vocalizations of the bush dog (Speothos venaticus), crab-eating fox (Cerdocyon thous), and maned wolf (Chrysocyon brachyurus) were studied in a captive setting. The three species' repertoires were similar and included whines, long-distance calls, and growls. The whines and growls varied in amplitude, duration, and repetition rate; these variations and the frequencies of occurrence probably were influenced by both social organization and habitat characteristics. The bush dog, which lives in family groups and inhabits rain forests, had an elaborate whine-scream continuum that at close distances conveyed a wide range of moods. On the other hand, the maned wolf, which is solitary and frequents open pampas, used amplitude variations of the growl to signal hostile intentions over short, medium, and long distances.The long-distance vocalizations of the three species were the least variable in structure; the bush dog and crab-eating fox used the call to promote contact with separated pairmates or family members. Based on recordings during development, these two calls appeared to arise from the repetitive whine, a variant of the basic whine syllable. The maned wolf also had a contact-promoting howl, apparently an extended whine of high amplitude, but it was rarely recorded during the present study. More prominent was the maned wolf roar-bark, which functioned to space individuals over large distances, and appeared to be a growl of high amplitude.     

Induction of fertile oestrus in the bitch using Deslorelin, a GnRH agonist

Oestrus induction in various canine breeds was attempted in 32 bitches. A group of 8 bitches were treated 80–160 d following their previous oestrus (G1) whereas a second group of 24 bitches (G2) were implanted 200–590 d following their previous oestrus. The treatment for each bitch consisted in one Deslorelin implant (Suprelorin®4,7mg, Virbac, France), inserted subcutaneously in the post-umbilical region. Ovulation, pregnancy rate and litter size were recorded. All bitches came in heat 4.3 ± 1.4 d after implantation (2–7 d). Ovulation was reported in 62.5% in G1 and 87.5% in G2. One bitch refused mating and since no AI was performed, she was not considered for further analysis. Pregnancy was obtained in 25% in G1 versus 78.3% in G2. Mean litter size was 6.7 ± 3.5 puppies (1–14). Luteal failure was suspected in 3 bitches, two that remained non-pregnant and one which aborted 58 d post-ovulation since the owner refused progesterone supplementation. Deslorelin implants can therefore be considered as a valuable alternative to induce fertile oestrus in bitches in anoestrus. Follow-up of the luteal phase is recommended, since some bitches might encounter luteal failure.     

Effect of an implant containing the GnRH agonist deslorelin on secretion of LH, ovarian activity and milk yield of postpartum dairy cows

Prevention of high plasma progesterone concentrations in the early postpartum period may improve fertility. Our objective was to determine whether a Deslorelin implant (DESL; 2100 microg, s.c.) would reduce secretion of LH and alter follicle dynamics, plasma concentrations of progesterone, estradiol and PGF2alpha metabolite (PGFM) in postpartum dairy cows. Cows received DESL on Day 7 postpartum (Day 7, n=8) or were untreated (Control, n=9). All cows were injected with GnRH (100 microg, i.m.) on Day 14 to assess LH response. A protocol for synchronization of ovulation with timed AI was initiated on Day 60 (GnRH [Day 60], CIDR [Day 60 to Day 67], PGF2alpha [Day 67, 25 mg and Day 68, 15 mg], GnRH [Day 69] , AI [Day 70]). The LH response to injection of GnRH on Day 14 was blocked in animals treated with DESL. Numbers of Class 1 (<6 mm) follicles were unaffected (P > 0.05) whereas numbers of Class 2 (6 to 9 mm) (P < 0.01) and Class 3 (>9 mm) follicles were less (P < 0.01) in DESL cows between Day 7 and Day 21. From Day 22 to Day 60, DESL-treated cows had more of Class 1 follicles and less Class 2 (P < 0.01) and Class 3 (P < 0.01) follicles, and lower plasma concentrations of progesterone and estradiol (P < 0.01). Concentrations of PGFM between Day 7 and Day 42 were not affected by treatment (P > 0.05). All cows ovulated in response to GnRH on Day 69. Subsequent luteal phase increases in plasma progesterone concentrations (Day 70 to Day 84) did not differ. The use of the DESL implant associated with PGF2alpha given 14 days later suppressed ovarian activity and caused plasma progesterone concentrations to remain < 1 ng/mL between Day 22 and Day 51. The DESL implant did not affect milk production.     

Endocrine changes,timing of ovulation,ovarian follicular growth and efficacy of a novel protocol (Estradoublesynch) for synchronization of ovulation and timed artificial insemination in Murrah buffaloes (Bubalus bubalis)

Experiments were conducted to investigate (1) ovarian follicular growth, timing of ovulation and associated endocrine changes (progesterone, estrogen, and LH) in cycling, and (2) efficacy in terms of pregnancy rate in cycling and anestrus Murrah buffaloes subjected to the Estradoublesynch protocol (prostaglandin F2α [PGF_2α] 0, GnRH 2, PGF_2α 9, estradiol benzoate, EB 10). Twelve cycling buffaloes were subjected to the Estradoublesynch protocol and observed for ovulation, follicle size, and endocrine changes after EB treatment. Ovulation occurred in 12 of 12 buffaloes (100%) at 48.5 ± 1.6 hours (range, 38.0–56.0) after EB treatment. Plasma LH, total estrogen, and progesterone concentrations were determined in intensive blood samples collected after EB treatment. Peak LH concentration of 34.2 ± 7.7 ng/mL (range, 17.8–178.5) occurred at 18.3 ± 0.8 hours (range, 14.0–22.0) after EB treatment. Peak total estrogen of 50.8 ± 6.9 pg/mL (range, 32.3–82.7) occurred 5.7 ± 1.0 hours (range, 2.0–14.0) after EB treatment. Follicle size was gradually increased from second PGF_2α injection (9.7 ± 0.3 mm; range, 8.0–12.0) until ovulation was detected (12.9 ± 0.4 mm; range, 11.0–15.0). Fourteen cycling and 11 anestrus buffaloes were subjected to the Estradoublesynch protocol, with timed artificial insemination (TAI) 48 and 60 hours after EB treatment, and 58 cycling buffaloes were inseminated after spontaneous estrus (control group). Pregnancy rates were 62% for TAI of cycling buffaloes, 64% for anestrus buffaloes, and 34.5% for control group. Our observations demonstrated that the Estradoublesynch protocol followed by TAI satisfactory enhanced pregnancy rates in both cycling and anestrus buffaloes.     

Assessment of ovarian activity in captive goral (Naemorhedus griseus) using noninvasive fecal steroid monitoring

To date, there is no information on gonadal steroidogenic activity of female goral (Naemorhedus griseus), a threatened species of Thailand. Captive goral populations have been established to produce animals for ex situ conservation and reintroduction, but as yet none are self-sustaining. The objectives of the present study were to (1) determine the influence of season on ovarian steriodogenic function; and (2) examine the relationship between gonadal hormone excretion and sexual behaviors throughout the year. Fecal samples were collected 5 to 7 days/wk for 15 months from 8 adult females housed at Omkoi Wildlife Breeding Center in Thailand and analyzed for ovarian steroid metabolites using validated enzyme immunoassays. Observations of sexual behaviors and mating were conducted each morning for 30 min/session. Based on fecal estrogen and progestagen metabolite concentrations, the overall estrous cycle length was about 21 days, with a 2- to 3-day follicular phase and an 18- to 20-day luteal phase. Sexual behaviors, most notably tail-up, increased for 2 to 3 days during the time estrogens were elevated during mating. Fecal progestagens were elevated during luteal phases and increased further during gestation, which lasted approximately 7 months. The lactation period was 5 months, and females were anestrus for 2 to 5 of those months, with the exception of one that cycled continuously throughout. Two females conceived around 2 months postpartum and so were pregnant during lactation. Birth records over the past 21 years indicated young are born throughout the year. This combined with the hormonal data suggests that female gorals are not strongly seasonal, at least in captivity, although there was considerable variation among females in estrogen and progestagen patterns. In conclusion, fecal steroid metabolite monitoring is an effective means of assessing ovarian function in this species and will be a useful tool for breeding management and planned development of assisted reproductive techniques such as artificial insemination and embryo transfer.     

Incorporation of a GnRH agonist,leuprolide acetate,into regimens with exogenous gonadotropins to produce ovarian stimulation and ovulation in the nonpregnant squirrel monkey

This study was designed to measure the effects of variations in the length of pretreatment with a GnRH agonist, leuprolide acetate (LA), on subsequent follicular development and ovulation. The hypothesis was that the duration of LA suppression of pituitary function does not adversely affect ovarian response to standardized ovulation induction protocols in squirrel monkeys. The first phase determined the dose and duration of LA needed to achieve a hypogonadal state. One of two groups received daily subcutaneous injections of 50 μg of LA. The other received a single injection of 175 μg of a depot suspension of LA. Sera were assayed for estradiol (E2) and progesterone (P). E2 and P levels increased 2‐ to 5‐fold with peak levels on days 4 and 7, respectively. Suppression of steroid levels took 10 to 15 days in the LA‐treated group. Depot‐LA did not effectively suppress steroid production. After suppression, females receiving daily LA received five daily injections of hMG to stimulate follicular development. E2 and P increased in these animals. These results suggest that cycling squirrel monkeys have P‐secreting capacity throughout the cycle. This may explain how the squirrel monkey is able to accommodate both a short (4–5 day) luteal phase of their 9 day cycle and implantation from 5 to 7 days after ovulation. A second study compared exogenous follicle stimulating hormone (FSH) to endogenous gonadotropins released as a response to LA in ovulation induction. Steroid production and hCG‐induced ovulation were assessed. LA treatment was compared to a standard ovulation induction protocol by using a randomized cross‐over measures design. There were no differences in E2 and P levels in response to dosages of either LA or hMG. The ovulatory response following LA treatment was not significantly greater than that using FSH. The number of animals with unovulated, large follicles was greater on the FSH protocol (12/18) compared to the LA protocol (3/18). Thus, a single injection of a depot preparation of LA is sufficient to stimulate follicular development and ovulation when followed by an hCG injection. Based on this observation and the data on unovulated large follicles, it is suggested that the ovary responds more readily to endogenous gonadotropins released by LA than to exogenous FSH. Am. J. Primatol. 49:153–164, 1999. © 1999 Wiley‐Liss, Inc.     

Establishment and characterization of an ovarian cell line of the silkworm, Bombyx mori

A cell line BmN-SWU1 was established from the ovarian tissues of 3-day-old fourth instar Bombyx mori larvae of the 21-872nlw variety by performing primary cultures in Grace's medium supplemented with 20% fetal bovine serum (FBS). The cell line primarily consisted of short spindle cells and round cells. The frequency of cells with chromosome number 2n = 56 was 80.5%; therefore, the cell line was considered to be a diploid cell line. The population-doubling time (PDT) at 45th passage line was 57.7 h. This cell line was susceptible to the B. mori nuclear polyhedrovirus (BmNPV), and the median tissue culture infective dose (TCID₅₀) at a cell density of 10⁵ cells/ml was 16.3 OBs/ml. The transient expression efficiency of the green fluorescent protein (GFP) gene in this cell line was 54.8%. We used the BmN-SWU1 cell line to select and establish a GFP transgenic cell line.     

Induction of lysozymelike activity in the hemolymph and hemocytes of an insect, Spodoptera eridania

Lysozymelike activity is present in the hemocytes and cell-free hemolymph of Spodoptera eridania. Its level remains essentially constant during larval development and can be induced by injection of various foreign materials. Serum bacteriolytic activity rises 24 hr after injection of saline, BSA, bacteria, bacterial endotoxin (LPS), latex particles, or sham injection. However, the magnitude and subsequent duration of the response depends on the nature of the injected material. The response is transient following sham injection or injection of soluble substances, such as saline and BSA, as compared to treatment with latex or bacteria. Both soluble and insoluble fractions of bacterial LPS preparations stimulated the lysozyme response. The response to a single injection of E. coli LPS was dose dependent and persisted for at least 5 days; however, additional injections had no effect on serum lysozyme level. The basal intracellular lysozyme level was significantly increased by E. coli LPS injection. Lysozyme release by hemocytes was proportional to intracellular concentration and did not increase after phagocytic stimulation of hemocytes.     

Comparison of endocrine changes,timing of ovulations,ovarian follicular growth,and efficacy associated with Estradoublesynch and Heatsynch protocols in Murrah buffalo cows (Bubalus bubalis)

Poor estrus expression and the difficulty encountered in predicting the time of ovulation compromise the reproductive efficiency of Murrah buffalo cows. Synchronization of ovulation and timed artificial insemination are able to precisely control the time of ovulation and thus avoid the need for estrus detection. Recently, the Estradoublesynch protocol (administration of a PGF2α injection 2 days before Heatsynch protocol; GnRH 0, PGF2α 7, estradiol benzoate [EB] 8) was developed that precisely synchronized ovulation twice, i.e., after GnRH and EB injections and resulted in satisfactory pregnancy rates in Murrah buffaloes. The present study was conducted on 104 cycling and 31 anestrus buffaloes to compare (1) the endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy of Estradoublesynch and Heatsynch protocols in cycling and (2) the efficacy of Estradoublesynch and Heatsynch protocols for the improvement of fertility in cycling and anestrus Murrah buffalo cows. Ovulation was confirmed after all GnRH and EB treatments by ultrasonographic examination at 2-hour intervals. Plasma progesterone and total estrogen concentrations were determined in blood samples collected at daily intervals, beginning 2 days before the onset of protocols until the day of second ovulation detection. Ovulatory follicle size was measured by ultrasonography at six time points (first PGF2α administration of Estradoublesynch protocol every 2 days before the onset of Heatsynch protocol, GnRH administration of both protocols, 2 hours before ovulation detection after GnRH administration of both protocols, second PGF2α injection of Estradoublesynch protocol, PGF2α injection of Heatsynch protocol, EB injection of both protocols and, 2 hours before ovulation detection after EB administration of both protocols). Plasma LH, total estrogen, and progesterone concentrations were determined in blood samples collected at 30-minute intervals for 8 hours, beginning GnRH and EB injections, and thereafter at 2-hour intervals until 2 hours after the detection of ovulation. The first ovulatory rate was significantly higher (P < 0.05) in the Estradoublesynch protocol (84.6%) than that in the Heatsynch protocol (36.4%). The first LH peak concentration (74.6 ± 10.4 ng/mL) in the Estradoublesynch protocol was significantly higher (P < 0.05) than that of the Heatsynch protocol (55.3 ± 7.4 ng/mL). In Estradoublesynch protocol, the total estrogen concentration gradually increased from the day of GnRH administration coinciding with LH peak, and then gradually declined to the basal level until the time of ovulation detection. However, in Heatsynch protocol, the gradual increase in total estrogen concentration after GnRH administration was observed only in those buffalo cows, which responded to treatment with ovulation. In both Estradoublesynch and Heatsynch protocols, ovulatory follicle size increased by treatment with GnRH and EB until the detection of ovulation. The pregnancy rate after the Estradoublesynch protocol (60.0%) was significantly higher (P < 0.05) than that achieved after the Heatsynch protocol (32.5%). Satisfactory success rate using the Estradoublesynch protocol was attributed to the higher release of LH after treatment with GnRH, leading to ovulation in most of the animals and hence creating the optimum follicular size at EB injection for ovulation and pregnancy to occur.     

Comparison of diet and feeding activity of two polychaetes, Nephtys caeca (Fabricius) and Nereis virens (Sars), in an estuarine intertidal environment in Québec, Canada

It is crucial to study how food resources are allocated in order to understand how invertebrate species of soft-bottomed benthic communities can live in the same environment, especially when belonging to the same trophic level. On the southern shore of the Lower St. Lawrence Estuary, the polychaetes Nereis virens (Nereidae) and Nephtys caeca (Nephtyidae) represent the main infaunal predators and are known to regulate (by predation) other populations within their community. For these animal populations, there is a high degree of spatial overlap in the horizontal dimension as well as vertically in the sediment column. The purpose of this study was to describe and compare the feeding activity of N. virens and N. caeca in relation to several environmental factors. The feeding activity of N. caeca was further studied under natural and experimental conditions and a feeding experiment was carried out to evaluate how the food resource is subdivided in conditions of high spatial overlap. Finally, the prospecting activity of N. virens was described under natural conditions. N. virens and N. caeca exhibited a high degree of dietary overlap, particularly at the adult stage. This food resource overlap can easily influence the regulation of these species, even though they display different feeding responses to environmental stimuli. Tides and storm events can also influence the feeding behaviour. We conclude that different behavioural feeding responses likely favour the occurrence of these two species in the same trophic environment.     

Genetic link between Cabeza, a Drosophila homologue of Fused in Sarcoma (FUS), and the EGFR signaling pathway

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease that causes progressive muscular weakness. Fused in Sarcoma (FUS) that has been identified in familial ALS is an RNA binding protein that is normally localized in the nucleus. However, its function in vivo is not fully understood. Drosophila has Cabeza (Caz) as a FUS homologue and specific knockdown of Caz in the eye imaginal disc and pupal retina using a GMR-GAL4 driver was here found to induce an abnormal morphology of the adult compound eyes, a rough eye phenotype. This was partially suppressed by expression of the apoptosis inhibitor P35. Knockdown of Caz exerted no apparent effect on differentiation of photoreceptor cells. However, immunostaining with an antibody to Cut that marks cone cells revealed fusion of these and ommatidia of pupal retinae. These results indicate that Caz knockdown induces apoptosis and also inhibits differentiation of cone cells, resulting in abnormal eye morphology in adults. Mutation in EGFR pathway-related genes, such as rhomboid-1, rhomboid-3 and mirror suppressed the rough eye phenotype induced by Caz knockdown. Moreover, the rhomboid-1 mutation rescued the fusion of cone cells and ommatidia observed in Caz knockdown flies. The results suggest that Caz negatively regulates the EGFR signaling pathway required for determination of cone cell fate in Drosophila.     

Ultrastructure of the spermatozoon of Anomotaenia quelea (Mettrick, 1961) (Cestoda,Cyclophyllidea, Dilepididae), an intestinal parasite of Quelea quelea (Aves,Ploceidae) in Senegal

The mature spermatozoon of Anomotaenia quelea exhibits an apical cone of electron-dense material and two helicoidal crest-like bodies. The apical cone near its base is surrounded by a lucent cytoplasm and a spiraled layer of cortical microtubules. The crest-like bodies are of different lengths, spiraled and make an angle of 30–40° to the hypothetical spermatozoon axis. The axoneme is of the 9 + ‘1’ trepaxonematan pattern and is surrounded by a periaxonemal sheath of electron-dense material. The cytoplasm contains in regions III and IV numerous electron-dense granules situated between the periaxonemal sheath and the cortical microtubules. The posterior extremity of the spermatozoon of A. quelea exhibits a nucleus and a disorganized axoneme and cortical microtubules. This type of posterior extremity of the mature spermatozoon has never been described previously in a Dilepididae. Similarly, two crest-like bodies have not been observed before in a dilepidid cestode.     

Anatomy and systematic affinity of Stanleya neritinoides (Smith, 1880), an enigmatic member of the thalassoid gastropod species flock in Lake Tanganyika, East Africa (Cerithioidea, Paludomidae)

The radiation of gastropods in Lake Tanganyika is an ideal system for testing competing hypotheses of species flock formation. Yet, much of the basic biology of these species remains unknown. In an ongoing effort to understand the evolution of Tanganyikan gastropods, we here describe Stanleya neritinoides. Alcohol‐preserved material of the soft parts is rare, consequently, the systematic position of the species, and a repeated suggested affinity to Tanganyicia rufofilosa, have been based primarily on features of the shell. However, features of the radula and operculum are unique and do not suggest an affinity to any other Tanganyikan species. Thus, S. neritinoides has remained a particularly poorly known and enigmatic member of the species flock. This investigation confirmed that several aspects of internal and external anatomy are shared between S. neritinoides, T. rufofilosa, and other Tanganyikan gastropods, but that S. neritinoides is unique in features of the radula and seminal receptacle. Moreover, S. neritinoides differs from T. rufofilosa in features of the foregut, midgut, hindgut, kidney, nervous system, reproductive system and reproductive strategy. These new data are inconsistent with an interpretation of identity of Stanleya and Tanganyicia. In addition, given the pervasive differences between the two, a sister‐group relationship between the two is unlikely. More precise systematic placement of S. neritinoides awaits the establishment of a phylogenetic framework for all Tanganyikan gastropods.     

Nest-site selection and reproductive success of a critically endangered parrot,the Great Green Macaw (Ara ambiguus), in an anthropogenic landscape

Nest-site selection is the principal way in which secondary cavity-nesting species mitigate the negative effects of factors such as predation, parasitism and exposure on reproductive success. Large-bodied secondary cavity-nesting birds rely on large cavities in mature trees that are often absent or reduced in anthropogenically disturbed forests. Hence, the availability of high-quality nest-sites may be limited for these species, potentially reducing reproductive success. The aim of this study was to investigate nest-site selection and the effect of nest-site features on reproductive success in the critically endangered Great Green Macaw Ara ambiguus in northern Costa Rica. We show that Great Green Macaws select nest-sites according to the characteristics of the cavity and the tree in which they are located. Moreover, reproductive success is a function of certain cavity features. However, the availability of high-quality cavities, those with features associated with higher reproductive success, is potentially limited. By studying nest-site selection and productivity together, we have identified that the availability of high-quality cavities could potentially limit the future recovery of the critically endangered Great Green Macaw.     

Agouti signalling protein is an inverse agonist to the wildtype and agonist to the melanic variant of the melanocortin-1 receptor in the grey squirrel (Sciurus carolinensis)

The melanocortin-1 receptor (MC1R) is a key regulator of mammalian pigmentation. Melanism in the grey squirrel is associated with an eight amino acid deletion in the mutant melanocortin-1 receptor with 24 base pair deletion (MC1RΔ24) variant. We demonstrate that the MC1RΔ24 exhibits a higher basal activity than the wildtype MC1R (MC1R-wt). We demonstrate that agouti signalling protein (ASIP) is an inverse agonist to the MC1R-wt but is an agonist to the MC1RΔ24. We conclude that the deletion in the MC1RΔ24 leads to a receptor with a high basal activity which is further activated by ASIP. This is the first report of ASIP acting as an agonist to MC1R.     

A diapause induction-termination asymmetry in the photoperiodic responses of the Linden bug, Pyrrhocoris apterus and an effect of near-critical photoperiods on development

The critical daylength for the induction of adult (ovarian) diapause in Pyrrhocoris apterus (central Bohemian population) was found to be 15.75 hr per 24 hr, whereas that for diapause termination was 1 hr longer, at 16.75 hr per 24 hr. The termination threshold remained unaltered through 28 to 98 days (4–14 weeks) in diapause, but the need for photoperiodic termination was abolished after 12–16 weeks at 4°C. The rate of nymphal development (25°C) was relatively rapid at both short (< 14.5 hr per 24 hr) and long (> 16.5 hr per 24 hr) photoperiods, but was significantly protracted at intermediate or ‘critical’ daylengths.     

Distyly and incompatibility in Psychotria homalosperma (Rubiaceae), an endemic plant of the oceanic Bonin (Ogasawara) Islands

Distyly rarely occurs on oceanic islands. Indeed, this form of polymorphism is absent on the Hawaiian Islands and in New Zealand. However, Psychotria homalosperma (Rubiaceae), which is endemic to the oceanic Bonin Islands of Japan, was presumed to maintain a distylous breeding system and floral morphology. We investigated the floral and reproductive characteristics of this species and examined incompatibility in self- and intramorph pollination. Psychotria homalosperma is morphologically and functionally distylous, based on findings with the examined populations, but its stigma and anther heights are not exactly reciprocal between the two morphs. This is the second example of distyly among the flora from the Bonin Islands, the other being the endemic P. boninensis. Both of these distylous species contrast with the Hawaiian Psychotria species, which have already become dioecious. The flowers of the long (L)-styled (pin) morph of the Bonin Islands plants are completely self- and intramorph incompatible, while the flowers of the short (S)-styled (thrum) morphs are partially intramorph compatible. The S-styled morph was more abundant in the examined populations from three islands of the archipelago, probably due to the existence of partial S–S intramorph compatibility.     

Effect of salinity variation and pesticide exposure on an estuarine harpacticoid copepod, Microarthridion littorale (Poppe), in the southeastern US

The harpacticoid copepod Microarthridion littorale (Poppe) was tested for interaction effects between salinity change and acute pesticide exposure on the survival and genotypic composition of a South Carolina population. Previous data suggested a significant link between a combined exposure to chlorpyrifos (CHPY) and dichloro-diphenyl-trichloroethane (DDT) and mitochondrial haplotype in the cytochrome b apoenzyme for this euryhaline species when exposed at 12-ppt salinity seawater. Our tests demonstrate a significant non-linear survival response for M. littorale to short-term immersion (24 h) in 3-, 12- and 35-ppt seawater, with copepods transferred to 12-ppt seawater having the lowest survival. There was significant statistical interaction between salinity and pesticide exposure for the dependent variable “survival.” However, changes in genetic composition of survivors were not significant, and they were complicated by extremely low survival in the pesticide/3 ppt and pesticide/36 ppt treatments. As noted for many studies of harpacticoids, males faired worse than females in all treatments, with none surviving pesticide exposure at 45 μg/l CHPY and 6 μg/l DDT.     

Assembly and annotation of full mitochondrial genomes for the corn rootworm species, Diabrotica virgifera virgifera and Diabroticabarberi (Insecta: Coleoptera: Chrysomelidae), using Next Generation Sequence data

Complete mitochondrial genomes for two corn rootworm species, Diabrotica virgifera virgifera (16,747 bp) and Diabrotica barberi (16,632; Insecta: Coleoptera: Chrysomelidae), were assembled from Illumina HiSeq2000 read data. Annotation indicated that the order and orientation of 13 protein coding genes (PCGs), and 22 tRNA and 2 rRNA sequences were in typical of insect mitochondrial genomes. Non-standard nad4 and cox3 stop codons were composed of single T nucleotides and likely completed by adenylation, and atypical TTT start codons was predicted for both D. v. virgifera and D. barberinad1 genes. The D. v. virgifera and D. barberi haplotypes showed 819 variable nucleotide positions within PCG regions (7.36% divergence), which suggest that speciation may have occurred ~ 3.68 million years ago assuming a linear rate of short-term substitution. Phylogenetic analyses of Coleopteran MtD genome show clustering based on family level, and may have the capacity to resolve the evolutionary history within this Order of insects.