Antioxidant Enzyme Activities Vary with Predation Risk and Environmental Conditions in Free-Living Passerine Birds.

Prolonged physiological stress response may lead to an excessive production of reactive oxygen species (ROS) and ultimately to oxidative stress and severe fitness costs. We investigated whether natural variation in predation risk, induced by pygmy owls (Glaucidium passerinum), modifies the oxidative status of two free-living food-supplemented passerine bird species-the great tit (Parus major) and the willow tit (Poecile montanus)-in March 2012 and 2013. Predation risk significantly affected antioxidant enzyme activities of willow tits. Antioxidant enzyme activities (principal component factor 2 [PC2] representing glutathione-S-transferase and superoxide dismutase activities) were higher in high predation risk areas in 2013 than in low predation risk areas in the same year. Higher enzyme activities may suggest higher ROS production in birds living under high predation risk. In addition, antioxidant enzyme activities (PC2) were also higher in high predation risk areas in 2013 than in high predation risk areas in the previous year, 2012. This may represent variation in the risk represented by pygmy owls, which is probably inversely related to the natural fluctuations in the densities of their main prey, voles. In willow tits, PC1 (representing catalase, total glutathione, the ratio of reduced to oxidized glutathione, and protein carbonylation) was not affected by perceived predation risk, nor were antioxidant levels or enzyme activities in great tits. Higher enzyme activities observed in willow tits suggest that predator presence can modify the antioxidant status of avian prey, but the response also seem to be influenced by other environmental characteristics, like harsh winter conditions.

Distribution and concentration of maternal progesterone in the yolk of Greater Rhea eggs (Rhea americana).

Progesterone is the most concentrated maternal yolk steroid characterized to date in birds; however, no information about it is available in ratite eggs. We collected freshly laid eggs from zoo-housed Greater Rhea females (Rhea americana) bred under similar rearing conditions during two breeding seasons to characterize concentration and distribution of maternal yolk progesterone. After high-performance liquid chromatography analysis, yolk hormone was measured using a commercial electrochemiluminescence immunoassay. Progesterone concentrations were found to vary significantly among the yolk layers, supporting a follicular origin for this steroid in Greater Rhea eggs. Additionally, highly similar mean absolute yolk progesterone concentrations were detected between 2013 and 2015 breeding seasons (1,332.98 ± 82.59 and 1,313.59 ± 85.19 ng/g, respectively). These values are also comparable to those found in some domestic carinate species. Findings suggest that at population level, when rearing conditions are similar, mean absolute yolk maternal progesterone concentrations also appear bounded. Future research on the factors and mechanisms that regulate progesterone deposition in Greater Rhea eggs is needed to better understand whether its levels depend on different rearing conditions.

Pre-hatch lung development in the ostrich.

We studied development of the ostrich lung using light microscopy as well as electron microscopy techniques. At E24, the lung comprised a few epithelial tubes, interspersed with abundant mesenchyme with scattered profiles of incipient blood vessels. Between E24 and E39, the epithelial thickness was reduced by 90% from 13.5 ± 0.41 µm to 1.33 ± 0.014 µm (mean ± SD, respectively). Atria were evident at E32, and by E35, the first portions of the blood-gas barrier (BGB) measuring 3.41 ± 1.12 µm were encountered. Gas exchange tissue was well formed by E39 with atria, infundibulae, air capillaries and a mature blood-gas barrier (BGB). BGB formation proceeded through the complex processes of secarecytosis and peremerecytosis, which entailed decapitation of epithelial cells by cutting or pinching off respectively and by E39, the BGB was thin at 2.21 ± 1.21 µm. Vascular remodeling by intussusceptive angiogenesis was a late stage process mediated by intraluminal pillars in the pulmonary vasculature.

Serum biochemical values of adult ostriches (Struthio camelus) anesthetized with xylazine, ketamine, and isoflurane.

Because of the size, speed, and powerful kicking ability of adult ostriches (Struthio camelus), chemical immobilization and general anesthesia are commonly used in these animals to provide veterinary care. To determine the serum biochemical values in ostriches anesthetized with xylazine and ketamine and isoflurane, 7 healthy adult ostriches (weighing 100 to 130 kg) were fasted for 12 hours. Each bird received an intramuscular injection of xylazine (4 mg/kg), followed 20 minutes later by an intravenous injection of ketamine (8 mg/kg), and were then intubated and maintained under anesthesia with isoflurane gas. Jugular blood samples were collected from each bird immediately before, and 20 minutes after, administration of xylazine; 10 and 20 minutes after the administration of ketamine; 10, 30, and 50 minutes after the delivery of isoflurane; and 20 and 50 minutes after discontinuing isoflurane. Concentrations of potassium and glucose increased significantly after the administration of ketamine, and that increase was maintained until the end of the study period. During recovery, levels of creatinine and amylase increased significantly. We conclude that administration of xylazine and ketamine and isoflurane anesthesia may affect the concentration of some serum biochemical values. These results should be taken into account when blood samples are evaluated in chemically immobilized or anesthetized ostriches.

Distribution and morphology of argyrophilic cells in the digestive tract of the African ostrich.

We used light microscopy to elucidate the morphological features of argyrophilic cells in the digestive tract of the African ostrich (Struthio camelus). The results indicated that argyrophilic cells were found to be distributed among the epithelial cells of the mucosa or glands throughout the digestive tract, except for the esophagus; two types of argyrophilic cells were found; i.e., closed-type cells and cells with triangular or elongated shapes and with their apical cytoplasmic process in contact with the lumen (open-type cells); the greatest number of argyrophilic cells was found in the proventriculus, and the argyrophilic cell density gradually decreased from the proventriculus to the rectum; Furthermore, the number of argyrophilic cells in the duodenum and ileum was higher than that in the jejunum. This text still combined the characteristics that the argyrophilic cells in digestive tract of ostrich maybe related to different digestive function of different region and the basis of their morphology to carry on a discussion. It was speculated that argyrophilic cells in the digestive tract may have both endocrine and exocrine functions.

The use of xylazine, ketamine, and isoflurane for induction and maintenance of anesthesia in ostriches (Struthio camelus).

To evaluate the use of xylazine/ketamine and isoflurane for the induction and maintenance of anesthesia in adult ostriches (Struthio camelus), 7 healthy adult ostriches (weight 100-130 kg) were deprived of food for 12 hours and then given an injection of xylazine (4 mg/kg IM), followed 20 minutes later by an injection of ketamine (8 mg/kg IV). After intubation, each bird was maintained on isoflurane anesthesia, and physiologic and hematologic parameters were measured. The respiratory rate and the systolic, diastolic, and mean blood pressures decreased significantly 10 minutes after delivery of isoflurane, and these decreases continued until the isoflurane was discontinued. Jaw and pedal withdrawal reflexes were useful indicators for evaluating muscle relaxation and depth of anesthesia in the ostriches while under general anesthesia. Recovery from anesthesia was relatively smooth, with minimal complications, and was complete at mean (SD) 50 +/- 24 minutes after discontinuing isoflurane. From these results, we concluded that induction of anesthesia with xylazine-ketamine followed by maintenance with isoflurane produced sufficient anesthesia for performing surgical operations with relatively smooth recovery in adult ostriches.

Follicle dynamics and its relation with plasma concentrations of progesterone, luteinizing hormone and estradiol during the egg-laying cycle in ostriches.

The aims of this study were (i) to describe the changes in the volume of large ovarian follicles (diameter >3 cm) during the 48 h egg laying cycle in farmed ostriches, and (ii) to quantify factors affecting the volume of the largest measured follicle and the plasma concentrations of progesterone (P(4)) and estradiol-17beta (E(2)beta). In eight egg-producing birds, which all ovulated during the study period, transcutaneous ultrasound scanning and blood sampling was performed at 3 h intervals. The average volume of the total number of visualized large follicles (V(total)), the largest measured follicle (V(F1)), the second largest follicle (V(F2)) and of all follicles smaller than F2 (V(F3-Fn)) were each higher before than after oviposition. V(total), V(F2) and V(F3-Fn) nearly doubled in the 24-h period before oviposition, while V(F1) remained at an equal, rather high level until oviposition. Immediately after oviposition V(total), as well as the volume of the other follicle categories, decreased within 6 h, i.e. around the moment of ovulation. By performing statistical analysis on the basis of linear mixed-effects modelling, we quantified that: (i) V(F1) was 13.2% higher before than after oviposition and increased with 6.5% when LH increased with 1 ng/ml; (ii) P(4) levels were 93.2% higher before than after oviposition and increased with 43.1% for every 3 h closer to oviposition; when LH and E(2)beta levels and V(F1) increased with 1 ng/ml, 10 pg/ml and 10 ml, respectively, P(4) increased with 116.6%, 50% and 6.1%; and (iii) E(2)beta levels were 35.6% higher before than after oviposition, increased with 2.7% for every 3 h closer to oviposition and increased with 14.6% when LH increased with 1 ng/ml. It is concluded that during the egg-laying cycle in ostriches: (i) follicular mass, as estimated by the volume of visualized follicles larger than 3 cm, increases before and decreases after ovulation, and (ii) follicular dynamics and its accompanying endocrine plasma hormone profiles during the egg-laying cycle in ostriches follow a pattern similar to that in chickens.

Ultrastructural features of the uterus in the sexually immature ostrich (Struthio camelus) during periods of ovarian inactivity and activity.

The ultrastructure of the surface epithelium and tubular glands of the uterus in the immature ostrich is described. In ostriches with inactive ovaries the uterus is lined by a non-ciliated simple columnar epithelium, with basally located heterochromatic nuclei. Scanning electron microscopy revealed that these non-ciliated cells have a dense microvillous cover. A simple columnar to pseudostratified columnar epithelium, comprised of non-ciliated and ciliated cells, lines the uterus in birds with active ovaries. The ciliated cells possess a wide luminal region, which contains a nucleus and various organelles. An accumulation of secretory granules was observed in the apical regions of the non-ciliated cells, as well as in a few ciliated cells. In addition to non-ciliated and ciliated cells, a cell type with rarefied cytoplasm was also identified. These cells appear to correspond to calcium secreting cells identified in other avian species. The results of this study indicate that, although uterine differentiation is present in immature ostriches with active ovaries, the production of secretory product appears to occur mainly in non-ciliated epithelial cells.

Changes in numbers of large ovarian follicles, plasma luteinizing hormone and estradiol-17beta concentrations and egg production figures in farmed ostriches throughout the year.

In this study we described and analysed changes in the numbers of large ovarian follicles (diameter 6.1-9.0 cm) and in the plasma concentrations of luteinizing hormone (LH) and estradiol-17beta (E(2)beta) in relation to individual egg production figures of farmed ostriches (Struthio camelus spp.) throughout one year. Ultrasound scanning and blood sampling for plasma hormone analysis were performed in 9 hens on a monthly basis during the breeding season and in two periods of the non-breeding season. Our data demonstrated that: (1) large follicles were detected and LH concentrations were elevated already 1 month before first ovipositions of the egg production season took place; (2) E(2)beta concentrations increased as soon as the egg production season started; (3) numbers of large follicles, LH and E(2)beta concentrations were elevated during the entire egg production season; and that (4) numbers of large follicles, LH and E(2)beta concentrations decreased simultaneous with or following the last ovipositions of the egg production season. By comparing these parameters during the egg production season with their pre-and post-seasonal values, significant differences were found in the numbers of large follicles and E(2)beta concentrations between the pre-seasonal, seasonal and post-seasonal period; while LH concentrations were significantly different between the seasonal and post-seasonal period. In conclusion, our data demonstrate that changes in numbers of large follicles and in concentrations of LH and E(2)beta closely parallel individual egg production figures and provide some new cues that egg production in ostriches is confined to a marked reproductive season. Moreover, our data provide indications that mechanism, initiating, maintaining and terminating the egg production season in farmed breeding ostriches are quite similar to those already known for other seasonal breeding bird species.

The relation between ultrasonographic observations in the oviduct and plasma progesterone, luteinizing hormone and estradiol during the egg laying cycle in ostriches.

In this study we investigated the temporal relationship between ovulation, egg formation, oviposition and the changes in plasma concentrations of progesterone, luteinizing hormone and estradiol-17beta during the egg laying cycle in farmed ostriches. In 10 egg-producing birds, transcutaneous ultrasound scanning was performed at 3h intervals and blood sampling at hourly intervals during a period of at least 48h (one egg laying cycle). In hens (n=8) that ovulated during the observational period, the ovulated egg was first detected 2h after oviposition; thus, ovulation occurred shortly after oviposition in all birds. During the period between two consecutive ovipositions, the developing egg remained for 9h in the proximal part (infundibulum, magnum or isthmus) and for 39h in the distal part of the oviduct (uterus). In ovulating hens, plasma progesterone concentrations showed a characteristic and consistent profile: from basal levels of around 0.1ng/ml concentrations started to increase 12h before oviposition, reached an average maximum of 3.5ng/ml at 3h before oviposition and returned to basal levels 3h and 30min after oviposition. Changes in plasma luteinizing hormone and estradiol-17beta concentrations showed comparable patterns of elevation and decline relative to the timing of oviposition and ovulation. However, variation in their individual basal concentrations was generally larger and peak values were less conspicuous than those of progesterone. In non-ovulating hens (n=2) neither progesterone, nor luteinizing hormone nor estradiol-17beta showed elevations to peak concentrations before oviposition. These data demonstrate that during the egg laying cycle of ostriches, events such as ovulation, egg development and oviposition evolve according to a rather strict time schedule, and that progesterone, luteinizing hormone and estradiol-17beta reach peak concentrations shortly before ovulation. Additionally, our findings also show that on-farm ultrasound scanning is a useful technique to discriminate between ovulating and non-ovulating hens.

Morphological and histological study of the ostrich (Struthio Camelus L.) liver and biliary system.

The peculiarity of the digestive system of the ostrich (Struthio Camelus L.), which is characterized by the continuous production of bile, led us to undertake macroscopical and histological studies of the liver and its biliary system, since very little bibliographic data exist on the subject. For this purpose we observed the organs of male and female ostriches 16-18 months of age, in situ, in order to describe their location, relationships and morphology. Samples of the liver were processed for observation by light microscopy; samples of the hepatoenteric duct were processed for observation by light and electron microscopy. Our findings regarding the liver revealed the presence of two lobes: a left lobe, subdivided into three lobes, and a right undivided lobe. There was no gall-bladder. The histological picture showed unlimited hepatic lobules, with hepatocytes arranged in cord-like fashion two cells thick. A large hepatoenteric duct arose from the porta hepatis, and opened into a papilla in the descending limb of the duodenum. The mucosa of the duct was lined by simple columnar epithelium consisting of cells having the same morphological cytoplasmatic features but distinguished by either a light or a dark nucleus.

The morphological features of the rete testis of the ostrich (Struthio camelus).

The epithelium of the rete testis of the ostrich is simple cuboidal to columnar in nature, unlike the more flattened epithelium encountered in other non-passerine birds. A solitary cilium projects into the ductal lumen. A unique lateral cell membrane modification, similar, in some respects, to a hemi-desmosome, occurs frequently along the length of this membrane. The lateral cell membrane is complexly folded, and the cell contains an abundance of intermediate filaments, particularly in the vicinity of the nucleus and the internal surface of the basal cell membrane. The Golgi complex and endoplasmic reticulum are moderately developed. Another unique feature of the cell is the presence of a solitary, large, heterogeneous lipid body/droplet situated in the immediate supranuclear region of the cell. Overall, the cell structure conveys the impression of a metabolically active cell, which has the capability of transporting material from the duct lumen to the basal part of the cell, and of protein synthesis, apparently much in excess of what is necessary for cellular maintenance and sustenance.

Progesterone and oestrogen receptor immunoreactivity in the vagina of the immature ostrich, Struthio camelus.

1. The presence of sperm storage tubules in the cranial, middle and caudal regions of the vagina in the immature ostrich was studied. In addition, progesterone and oestrogen receptor immunoreactivity in these regions was investigated. 2. Sperm storage tubules were observed only in birds with active ovaries. Progesterone receptor immunoreactivity was more intense in birds with active ovaries, whilst immunostaining for the oestrogen receptor was absent in birds with both active and inactive ovaries. 3. These results suggest that steroid hormones produced by the active ovary mediate the activation of the progesterone receptor.