Culex quinquefasciatus (Diptera: Culicidae) survivorship following the ingestion of bird blood infected with Haemoproteus sp. parasites.

Arthropod vectors are frequently exposed to a diverse assemblage of parasites, but the consequence of these infections on their biology and behavior are poorly understood. We experimentally evaluated whether the ingestion of a common protozoan parasite of avian hosts (Haemoproteus spp.; Haemosporida: Haemoproteidae) impacted the survivorship of Culex quinquefasciatus (Say) (Diptera: Culicidae). Blood was collected from wild northern cardinals (Cardinalis cardinalis) in College Station, Texas, and screened for the presence of Haemoproteus spp. parasites using microscopic and molecular methods. Experimental groups of Cx. quinquefasciatus mosquitoes were offered Haemoproteus-positive cardinal blood through an artificial feeding apparatus, while control groups received Haemoproteus-negative cardinal blood or domestic canary (Serinus canaria domestica) blood. Culex quinquefasciatus mosquitoes exposed to Haemoproteus infected cardinal blood survived significantly fewer days than mosquitoes that ingested Haemoproteus-negative cardinal blood. The survival of mosquitoes fed on positive cardinal blood had a median survival time of 18 days post-exposure and the survival of mosquitoes fed on negative cardinal blood exceeded 50% across the 30 day observation period. Additionally, mosquitoes that fed on canary controls survived significantly fewer days than cardinal negative controls, with canary control mosquitoes having a median survival time of 17 days. This study further supports prior observations that Haemoproteus parasites can be pathogenic to bird-biting mosquitoes, and suggests that Haemoproteus parasites may indirectly suppress the transmission of co-circulating vector-borne pathogens by modulating vector survivorship. Our results also suggest that even in the absence of parasite infection, bloodmeals from different bird species can influence mosquito survivorship.

Characteristics of song, brain-anatomy and blood androgen levels in spontaneously singing female canaries.

Females of many northern temperate songbird species sing sporadically. However, detailed descriptions of female song are rare. Here we report a detailed analysis of song in a small number of spontaneously-singing female domesticated canaries (Serinus canaria) under non-breeding, laboratory conditions in a large population of domesticated birds. In-depth analysis showed that these females sang rarely, and the spontaneous songs varied between and within birds over time. Furthermore, spontaneous female songs were distinct from songs of testosterone-induced singing female canaries and from songs of male canaries in both temporal and spectral features. Singing females had significantly elevated plasma androgen levels and a larger size of the major song controlling brain nuclei HVC (used as a proper name) and the robust nucleus of the arcopallium (RA) than non-singing females housed under similar conditions. The sporadically observed production of song and accompanying differences in brain anatomy in female canaries may thus depend on minute intraspecific differences in androgen levels.

Eggshell Biliverdin and Protoporphyrin Pigments in a Songbird: Are They Derived from Erythrocytes, Blood Plasma, or the Shell Gland?

Biliverdin and protoporphyrin pigments are deposited into the eggshell when the developing egg is in the shell gland. However, the site of synthesis of eggshell pigments is still uncertain, although it may influence the possible costs and potential functions of eggshell coloration in avian species. Eggshell pigments may be derived from red blood cells or be produced in other organs and then transferred to the shell gland, or they may be synthesized de novo in the shell gland. We studied in the canary (Serinus canaria) whether eggshell blue-green and brown pigmentations are associated with experimentally elevated anemia, female hematocrit level, immature erythrocyte percentage, and feces and plasma pigment levels during egg laying to find out the possible origin of eggshell pigments. We found no significant effects of hematocrit level or experimentally elevated anemia on intensity of eggshell blue-green and brown pigmentations; therefore, we consider it less likely that eggshell pigments are derived from erythrocytes. In addition, we found no significant associations between female feces biliverdin concentration during egg laying and intensity of eggshell blue-green pigmentation, suggesting that eggshell biliverdin may not originate from the spleen or liver. We found a negative association between plasma and feces protoporphyrin concentrations during egg laying and eggshell brown chroma. This result suggests that an increased production of protoporphyrin in the liver, which could have elevated plasma and feces protoporphyrin concentrations, could inhibit eggshell protoporphyrin pigmentation, probably through affecting enzymatic activities. We suggest that both pigments are produced de novo in the shell gland in the canary, but circulating pigment levels may influence shell gland pigment synthesis, thus connecting the physiological status of the female to eggshell coloration.

Studies of HVC Plasticity in Adult Canaries Reveal Social Effects and Sex Differences as Well as Limitations of Multiple Markers Available to Assess Adult Neurogenesis.

In songbirds, neurogenesis in the song control nucleus HVC is sensitive to the hormonal and social environment but the dynamics of this process is difficult to assess with a single exogenous marker of new neurons. We simultaneously used three independent markers to investigate HVC neurogenesis in male and female canaries. Males were castrated, implanted with testosterone and housed either alone (M), with a female (M-F) or with another male (M-M) while females were implanted with 17ß-estradiol and housed with a male (F-M). All subjects received injections of the two thymidine analogues, BrdU and of EdU, respectively 21 and 10 days before brain collection. Cells containing BrdU or EdU or expressing doublecortin (DCX), which labels newborn neurons, were quantified. Social context and sex differentially affected total BrdU+, EdU+, BrdU+EdU- and DCX+ populations. M-M males had a higher density of BrdU+ cells in the ventricular zone adjacent to HVC and of EdU+ in HVC than M-F males. M birds had a higher ratio of BrdU+EdU- to EdU+ cells than M-F subjects suggesting higher survival of newer neurons in the former group. Total number of HVC DCX+ cells was lower in M-F than in M-M males. Sex differences were also dependent of the type of marker used. Several technical limitations associated with the use of these multiple markers were also identified. These results indicate that proliferation, recruitment and survival of new neurons can be independently affected by environmental conditions and effects can only be fully discerned through the use of multiple neurogenesis markers.

Does reproduction protect against oxidative stress?

A central principle of life-history theory is that parents trade investment in reproduction against that in body maintenance. One physiological cost thought to be important as a modulator of such trade-off is oxidative stress. Experimental support for this hypothesis has, however, proved to be contradictory. In this study, we manipulated the nestling rearing effort of captive canaries (Serinus canaria) soon after the hatching of their nestlings using a brood-size manipulation to test whether an increase in nestling rearing effort translates into an increase in oxidative damage, an increase in ceruloplasmin (which is upregulated in response to oxidative damage) and a decrease in thiol antioxidants. We also compared the blood oxidative stress level of reproducing birds with that of non-reproducing birds, a crucial aspect that most studies have invariably failed to include in tests of the oxidative cost of reproduction. Compared with non-breeding canaries and pre-manipulation values, plasma oxidative damage (reactive oxygen metabolites and protein carbonyls) decreased in breeding canaries irrespective of sex and brood size. In contrast, oxidative damage did not change in non-breeding birds over the experiment. Ceruloplasmin activity in plasma and both non-protein and protein thiols in red blood cells did not change throughout the experiment in both treatment groups. Our results suggest that reproduction may result in decreased rather than increased blood oxidative stress. Our results may explain some of the inconsistencies that have so far been reported in experimental tests of the oxidative cost of reproduction hypothesis.

A new method for in vitro detection of bromodeoxyuridine in serum: a proof of concept in a songbird species, the canary.

Systemic injection of a thymidine analogue such as bromodeoxyuridine (BrdU) in vertebrates is commonly used to detect and study cell production during development, adulthood, and pathology, particularly in studies of adult neurogenesis. Although researchers are applying this technique to multiple species in various physiological conditions, the rate of BrdU clearance from the serum remains unknown in most cases. Changes in this clearance rate as a function of the species, sex or endocrine condition could however profoundly affect the interpretation of the results. We describe a rapid, sensitive, but simple bioassay for post-injection detection and quantification of BrdU in serum. This procedure was shown to be suitable for determining the length of time a thymidine analogue remains in the bloodstream of one avian species and seems applicable to any vertebrate provided sufficiently large blood samples can be collected. This technique was used to demonstrate that, in canaries, BrdU injected at a dose of 100 mg/kg is no longer available for incorporation into DNA between 30 and 60 min post-injection, a delay shorter than anticipated based on the available literature. Preliminary data suggest a similar fast clearance in Japanese quail and mice.

Modulation by steroid hormones of a "sexy" acoustic signal in an Oscine species, the Common Canary Serinus canaria.

The respective influence of testosterone and estradiol on the structure of the Common Canary Serinus canaria song was studied by experimentally controlling blood levels of steroid hormones in males and analyzing the consequent effects on acoustic parameters. A detailed acoustic analysis of the songs produced before and after hormonal manipulation revealed that testosterone and estradiol seem to control distinct song parameters independently. The presence of receptors for testosterone and estradiol in the brain neural pathway controlling song production strongly suggests that the observed effects are mediated by a steroid action at the neuronal level.

Developmental changes and among-sibling variation of corticosterone levels in an altricial avian species.

The postnatal development of the activity of the brain-pituitary-adrenal axis was investigated in an altricial bird species by measurements of plasma corticosterone levels in nestling and fledgling canaries, Serinus canaria. Corticosterone was detectable (>2.6 ng/ml) in 30% of 5-day-old, 67% of 10-day-old, 72% of 15-day-old, and 88% of 23-day-old birds. When detectable, the corticosterone levels of 5-day-old nestlings were comparable to the baseline levels of adult birds. Levels were higher in 10- and 15- than in 5-day-old nestlings. The levels of 23-day-old fledglings (about 6 to 7 days after fledgling) were significantly higher than those of 15-day-old nestlings. They were intermediate between adult baseline and stress-induced levels. Sex did not influence this general profile, but levels varied with the order of hatching within broods. At the age of 15 and 23 days first hatched chicks had higher corticosterone levels than last hatched chicks, while second hatched chicks had intermediate levels. These differences were not correlated with body mass. The results suggest that (1) the brain-pituitary-adrenal axis of this altricial bird becomes fully functional after hatching and (2) birth order within broods influences corticosterone secretion during subsequent stages of development. It is unlikely that the brain-pituitary-adrenal axis matures at different rates in first and later hatched chicks or that the different levels of first and later hatched chicks were caused by capture and handling stress. Rather, they may result from such maternal effects as hatching asynchrony or differential concentrations of yolk steroids among the eggs in a clutch. Further studies will have to show whether this systematic variation of corticosterone levels among siblings during early life persists into adulthood and how it is related to behavior and fitness.

Neurogenesis in adult canary telencephalon is independent of gonadal hormone levels.

Neurons generated in adulthood are found throughout the canary telencephalon. We are interested in the factors that control the rate of proliferation of stem cells that give rise to these new neurons. The rate of incorporation of newly generated neurons into vocal-control regions varies seasonally. This difference could reflect a higher rate of neurogenesis, a lower rate of cell death, or an altered migration. We examined the incidence of thymidine-labeled cells in the telencephalic ventricular zone of adult canaries as a function of variations in gonadal hormone levels. Adult female canaries maintained on a short-day photoperiod were anesthetized and gonadectomized. Four separate groups of birds received systemic exposure to either testosterone, estradiol, a combination of an anti-androgen and an inhibitor of estrogen synthesis, or nothing. All birds were also implanted with an osmotic minipump that released 3H-thymidine for 3 d and were killed 4 or 7 d following the onset of treatment. Analysis of autoradiograms revealed no differences between groups in the incidence of labeling within the ventricular zone either at the level of the anterior commissure or directly adjacent to the vocal-control nucleus HVC (higher vocal center). These results suggest that sex steroids do not regulate the rate of cell division in the ventricular zone. Seasonal differences in the incorporation of labeled cells into HVC may therefore be due to regulation of neurogenesis by photoperiodic factors other than gonadal steroids or to some other cellular mechanism, such as differential migration or survival of neurons.

The Leucocytozoidae of South African birds: Passeriformes.

The leucocytozoids of ten families of Passeriformes--Estrildidae, Fringillidae, Laniidae, Nectariniidae, Passeridae, Ploceidae, Promeropidae, Pycnonotidae, Sturnidae and Zosteropidae--are reviewed. Leucocytozoon roubaudi from the Estrildidae, L. fringillinarum from the Fringillidae, L. balmorali from the Laniidae, L. gentili from the Passeridae, L. bouffardi from the Ploceidae, L. brimonti from the Pycnonotidae and L. zosteropis from the Zosteropidae are re-described. Leucocytozoon dutoiti, L. nectariniae, L. deswardti, L. pycnonoti and L. sturni are new species described from the Fringillidae (Carduelinae), Nectariniidae, Promeropidae, Pycnonotidae and Sturnidae respectively while L. monardi is considered to be a synonym of L. gentili and L. molpastis is considered to be a synonym of L. brimonti.

Seasonal changes in gonadal hormone levels of adult male canaries and their relation to song.

Samples of song and blood levels of three gonadal hormones, T, DHT, and E2, were taken at monthly intervals from six adult male canaries over a period of 1 year, as these birds went from 12 to 24 months of age. Song variability and addition of new syllable types were maximal during the summer and early fall, with a peak in September. A secondary peak in new syllable acquisition occurred in March. Blood T levels were particularly low during July-August and during February. Thus, a lowering of blood T levels preceded by about 1 month the two marked peaks in new syllable acquisition. Blood levels of the two other hormones were related less obviously to song learning peaks. It is hypothesized that a concurrence of hormonal, neural, and behavioral changes facilitates song learning in adulthood.

Inverse changes in plasma prolactin and LH concentrations in female canaries after disruption and reinitiation of incubation.

Prolactin and LH concentrations were measured in the plasma of female canaries sampled during the breeding cycle and after disruption and reinitiation of incubation behaviour. The late incubation period was characterized by low LH and high prolactin concentrations, and canaries separated from their nests at this stage showed an increased in LH and a decline in prolactin within 3 h. In one experiment mean (+/- S.E.M.) concentrations before and 24 h after nest deprivation were: prolactin 397 +/- 86 and 18 +/- 5 micrograms/l; LH 1.04 +/- 0.21 and 2.03 +/- 0.17 micrograms/l. Female canaries which abandoned their nests after the eggs had been removed also showed an increase in LH together with a fall in prolactin 24 h after egg removal. When nest-deprived canaries were allowed to resume incubation, plasma prolactin increased again within 5 h and after 2 days had reached levels normal for incubating birds (398 +/- 46 micrograms/l). Concentrations of LH changes more gradually but had decreased 2 days after the resumption of incubation. Thus prolactin and LH show inverse changes after the disruption and reinitiation of incubation behaviour; it is not clear, however, if the change in one is dependent on the other or if both hormones are responding to the same external stimulus.

Plasma concentrations of prolactin during incubation and parental feeding throughout repeated breeding cycles in canaries (Serinus canarius).

Concentrations of prolactin were measured in the plasma of male and female canaries serially sampled during repeated breeding cycles. Concentrations in female canaries were low during nest building and increased significantly (P less than 0.05) during the first few days of incubation. Levels increased further (P less than 0.01) in the middle of the incubation period to reach concentrations which were ten times higher than before breeding. Levels remained high for several days after the eggs hatched, as the young were fed in the nest. Prolactin concentrations declined gradually as the young were reared, reaching basal levels by the time the young were fledged, but always increased again in the females as they began incubating eggs in subsequent breeding cycles. Male canaries, which do not incubate but do assist in feeding the young, showed only slight increases in prolactin during the parental period. Male and female canaries which did not breed had low levels of prolactin throughout the experiment. The results show that prolactin secretion is high not only during incubation but also during the parental feeding phase in an altricial passerine bird. These findings are discussed in relation to the different patterns of prolactin secretion which occur during breeding in other birds.