A proteomic analysis of Nipponia nippon (ID#162)

We investigated the proteome of a female Crested Ibis (Nipponia nippon, ID#162) that died on March 10, 2010 at the Sado Japanese Crested Ibis Conservation Center. Protein preparations from the brain, trachea, liver, heart, lung, proventriculus, muscular stomach, small intestine, duodenum, ovary and neck muscle were subjected to in‐solution shotgun mass spectrometry (MS)/MS analyses using an LTQ Orbitrap XL mass spectrometer. A search of the National Center for Biotechnology Information Gallus gallus databases revealed 4253 GI (GenInfo Identifier) numbers with the sum of the same 11 tissues examined in the Crested Ibis. To interpret the obtained proteomics data, it was verified in detail with the data obtained from the brain of the Crested Ibis. It has been reported that drebrin A is specifically expressed in adult chicken brain. In the shotgun proteomic analyses of the Crested Ibis, we identified drebrin A as a brain‐specific protein. Furthermore, Western blotting analysis of the protein preparations from 10 tissues of the Crested Ibis and 150‐day‐old hens using anti‐drebrin antibodies showed intensive expression of approximately 110 kDa polypeptides of drebrin in both brains. We believe firmly that the present data will contribute to initial and fundamental steps toward understanding the Crested Ibis proteome.     

The W‐ and Z‐linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island

The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR‐based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W‐ and Z‐linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR‐based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR‐based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.     

Failure to turn eggs during incubation: effects on embryo weight, development of the chorioallantois and absorption of albumen

Turning eggs during incubation is essential for good hatchability. In the present paper additional effects on the development of the chorioallantois, absorption of albumen and growth of the embryo are recorded. The ability of an unturned egg to hatch was not affected by egg weight, egg shell porosity or water loss during incubation. The ability of the chorioallantois to spread around the inner surface of the inner shell membrane and the degree of absorption of the residual albumen affected the growth of the embryo and its ability to hatch. Unturned eggs hatched later than eggs which were turned throughout incubation.     

The interaction between maternal and post‐hatch n‐3 fatty acid supplementation in broiler diets

This study investigated whether offspring from n‐3‐supplemented breeders have an enhanced performance and immune organ weight when fed a post‐hatch n‐3‐enriched diet in comparison with their control‐fed counterparts and the importance of timing of omega‐3 supplementation. Therefore, 480 Ross‐308 broiler breeder hens were fed one of four different diets (120/treatment). The control diet (CON) was a basal diet, rich in n‐6 fatty acids (FA). The three other diets were enriched in n‐3 FA, formulated to obtain a different EPA/DHA ratio of 1/1 (EPA = DHA), 1/2 (DHA) or 2/1 (EPA). At 33 weeks of age, eggs were incubated to obtain 1440 offspring. They were set up according to their maternal diet and sex in 48 pens of 30 chicks each (12 pens per maternal treatment: six male and six female). Half of the offspring were given a post‐hatch control diet, whereas to other half received an n‐3‐supplemented diet. Zootechnical performance was followed for starter, grower and finisher phase, and at the end of each phase two, chicks per pen were sacrificed to determine the weight of the immune organs. No interaction was found between maternal and post‐hatch n‐3 treatment for zootechnical performance. An interaction arose between the maternal and post‐hatch n‐3 supplementation for proportional bursa weight at day 1 and day 14 and proportional liver weight at day 14, but effects on immune organ weight were rather limited. Offspring post‐hatch n‐3 supplementation did not enhance maternal n‐3 supplementation.     

笼养朱鹮非繁殖期昼间行为活动时间分配

采用即时取样和扫描法对河南董寨国家级自然保护区3个朱鹮笼舍的3对朱鹮行为进行了45 d,计585 h的观察。结果显示：（1）朱鹮非繁殖期昼间各行为休息（40.12%）占比例最大,其次是理羽（17.45%）、刨啄（14.03%）、运动（12.32%）、取食（11.30%）、剧烈运动（3.99%）、水浴（0.54%）、其他（0.26%）。（2）朱鹮活动呈一定的节律性,休息的2个显著高峰分别在6：00和18：00,另外2个小高峰分别在10：00和14：00;运动小高峰在12：00;取食2个显著高峰分别在9：00和15：00;理羽有4个小高峰,分别在7：00、10：00、14：00和17：00;水浴的2个高峰分别在13：00和14：00;刨啄的2个高峰分别在12：00和16：00。（3）两性昼间行为峰值也存在着一些差异,雌鸟和雄鸟相比之下,运动早,中午休息少,洗浴早,创啄多,取食强度较小。     

Effect of parental origin on early life history traits of European eel

Establishment of European eel (Anguilla anguilla) hatchery production will rely on selectively bred individuals that produce progeny with the best traits in successive generations. As such, this study used a quantitative genetic breeding design, between four females and nine males (four wild‐caught and five cultured), to investigate the effect of paternal origin (wild‐caught vs. cultured) and quantify the relative importance of parental effects, including genetic compatibility, on early life history (ELH) performance traits (i.e. fertilization success, embryonic survival at 32 hr post‐fertilization, hatch success and larval deformities at 2 days post‐hatch) of European eel. Wild‐caught males had higher (56%) spermatocrit values than cultured males (45%), while fertilization success, embryonic survival, hatch success and larval deformities were not significantly impacted by paternal origin. This demonstrates that short‐term domestication of male eels does not negatively affect offspring quality and enables the consideration of cultured male broodstock in future breeding programmes. Moreover, paternity significantly explained 9.5% of the variability in embryonic survival, providing further evidence that paternal effects need to be taken into consideration in assisted reproduction protocols. Furthermore, maternity significantly explained 54.8% of the variation for fertilization success, 61.7% for embryonic survival, 88.1% for hatching success and 62.8% for larval deformities, validating that maternity is a major factor influencing these “critical” ELH traits. At last, the parental interaction explained 12.8% of the variation for fertilization success, 8.3% for embryonic survival, 4.5% for hatch success and 20.5% for larval deformities. Thus, we conclude that eggs of one female can develop more successfully when crossed with a compatible male, highlighting the importance of mate choice for successful propagation of high‐quality offspring. Together, this knowledge will improve early offspring performance, leading to future breeding programmes for this critically endangered and economically important species.     

A simple and efficient method for extraction of PCR-amplifiable DNA from chicken eggshells

Recently, we reported a method for discriminating a Japanese brand of chicken, the Hinai-jidori. As an application of this method for discriminating Hinai-jidori eggs, we here report an efficient method for extracting maternal DNA from eggshells. Eggshell powder was completely decalcified with EDTA solution, and then DNA was isolated by conventional phenol-chloroform extraction and ethanol precipitation. The efficiency of DNA recovery from eggshells was 50-fold higher than that of a previously reported method. The recovered DNA could be used for PCR, and 10 markers for identifying the Hinai-jidori chicken were detected. The genotypes of the Hinai-jidori exactly matched those of the Hinai-dori breed. Using this method, Hinai-jidori and Hinai-dori eggs could be distinguished from the eggs of Rhode Island Reds. This is the first report of a technique that can be used to discriminate the eggs of Hinai-jidori from those of other chickens, and it can also be utilized to validate the labeling of Hinai-jidori eggs in the market.     

Clinical and pathological findings in two Mauritian pink pigeons (Columba mayeri)

Two Mauritian pink pigeons (Columba mayeri) from a captive breeding programme, with locomotor disorders were examined clinically and post mortem. One bird was unable to fly owing to a permanent luxation of the left elbow. The other bird had an inclined foot caused by a medial curvature and outward rotation of the tarsometatarsal bone, a deformity similar to the previously recorded 'inclined feet' in birds bred in Mauritius.     

Effect of pre-incubation storage conditions on hatchability,chick weight at hatch and hatching time in broiler breeders

1. Eggs were stored for two different times at varying temperatures. The effects on hatchability, chick weight at hatch and hatching time were examined in two broiler breeder lines from 33 to 58 weeks of age. 2. Short storage (1 to 3 d). Storage at 20 degrees C compared with 16.5 degrees C reduced hatchability of all eggs set. No effect was observed on hatchability of fertile eggs, hatching time or chick weight. 3. Long storage (9 to 11 d). Storage at 16.5 degrees C compared with 10 degrees C decreased both hatchability of fertile eggs and chick weight at hatch. Incidence of early embryonic death increased and incubation time decreased at 16.5 degrees C compared with 10 degrees C. 4. Chicks from morning eggs were heavier than those from afternoon eggs irrespective of storage conditions. 5. Hatchability (all eggs set and fertile eggs) and chick weight varied with hen age irrespective of storage conditions. During long storage, hatching time varied with hen age independently of breeder line, storage temperature or egg laying time. 6. Hatchability (all eggs set and fertile eggs) was higher in line A than in line B. Line B eggs hatched later and produced heavier chicks than line A eggs irrespective of storage time.     

Effects of in ovo feeding of L‐arginine on the development of digestive organs,intestinal function and post‐hatch performance of broiler embryos and hatchlings

This study was to investigate the effects of in ovo feeding (IOF) L‐arginine (Arg) solution on the development of digestive organs, the duodenal mucosa of broiler embryos and hatchlings, and the growth performance of chicks during the first week post‐hatch. A total of 720 fertilized eggs with similar weight were randomly allocated to three groups, consisting of eight replicates of 30 eggs each. Three treatments were arranged as non‐injected control, diluent‐injected (0.75% NaCl solution) group and Arg solution‐injected group containing 1% Arg, dissolved in diluent. At 17.5 days of incubation, 0.6 ml of IOF solution was injected into amniotic fluid of each egg of injected groups. Results showed IOF of Arg solution increased (p < .05) the chick embryo weight at 19 days of incubation; the body weight gain of post‐hatch broilers during 1–7 days; the weights of liver, pancreas, proventriculus and gizzard; the concentrations of duodenal ghrelin, vasoactive intestinal peptide and glucagon‐like peptide 2; and the duodenum mucosal enzyme activities of alkaline phosphatase, maltase, sucrase and inducible nitric oxide synthase of 7‐day‐old post‐hatch broilers compared with other groups. The IOF of Arg solution also increased (p < .05) the villus height (VH) and the ratio of VH to crypt depth (CD) and decreased (p < .05) the CD in duodenum of broiler embryos and post‐hatch hatchlings, except for the CD at 19 days of incubation. In conclusion, IOF of 1% Arg solution into the amnion at 17.5 days of incubation could improve the development of digestive organs, the duodenal morphology, the releasing of gastrointestinal hormones and mucosal enzyme activities of broiler embryos and hatchlings and finally the growth performance of chicks during the first week post‐hatch. Therefore, IOF of appropriate Arg solution could be an effective technology for regulating early nutrition supply and subsequent growth development in poultry industry.     

Genetic profile and serosurvey for virus infections of Japanese wild boars in Shikoku Island

To estimate the genetic background of Japanese wild boars inhibiting the 4 prefectures of Shikoku Island (Kagawa, Tokushima, Kouchi and Ehime), we examined haplotypes of mitochondrial DNA (mtDNA) and genotypes of the nuclear glucosephosphate isomerase-processed pseudogene (GPIP) in 189 wild boars. Of the 8 different mtDNA haplotypes (J5, J10, J12, J13, J15, J19, J20 and E33) detected in Shikoku Island, the first to be identified were J19 and J20 (of Japanese wild boar lineage) and E33 (of European domestic pig lineage). The presence of haplotype E33 indicates local crossbreeding between wild boar and escaped domestic pigs and/or crossbred Inobuta. Four kinds of the GPIP genotypes were detected from 189 wild baors in Shikoku Island, but no European GPIP alleles were found. In 113 wild boars, no antibodies against Classical Swine Fever or Aujeszky's disease were detected, suggesting that they had not been exposed to those pathogens.     

Fertility and hatchability results with fowl spermatozoa stored in fresh and freeze-dried diluent

Semen obtained from Rhode Island Red males was stored for 3 and 24 h in fresh and freeze‐dried diluent. Fertility and hatchability of eggs laid from the 2nd to 8th d inclusive after a single insemination was tested with hens of proven fertility. The fertilising power of diluted semen after storage did not differ significantly from the undiluted control, but the hatchability of the fertile eggs was significantly reduced after 24 h storage in the freeze‐dried diluent.     

一例朱鹮雏鸟皮下气肿的诊治

动物皮下气肿成因复杂,鸟类(禽类)的皮下气肿常见于体内气囊破裂所致。德清县珍稀动物繁育研究中心2012年首次出现了1例人工饲养的朱鹮雏鸟在15日龄时发生皮下气肿。该气肿位于右侧大腿处,内部充气,气泡内未见其他病变。根据其临床症状,推测可能是由于患雏与其他雏鸟嬉戏打斗过度的充气,或受到撞击等原因使气囊破裂。诊断为右侧腹或后胸气囊破裂,引发皮下气肿。鉴于临床上穿刺排气效果不显著,以及开创排气易引起细菌感染等因素,本病例采用了自然恢复的治疗方式,将患雏转入安静环境,避免外界干扰,加强防护,减少剧烈运动,实行隔离饲养等措施,15日后气肿变小,25日后气肿消失。该病例的诊治及病因的推测,为珍稀鸟类临床上该病的防治提供了方法。     

Risk Factors Associated with Salmonella Status of Broiler Flocks Delivered to Grow‐Out Farms

In a prospective field observational study in the southeastern USA, we sampled gastrointestinal (GI) tracts from chicks of 65 broiler flocks delivered to conventional grow‐out farms for rearing. The flocks were hatched at seven broiler hatcheries. The mean within‐flock prevalence of Salmonella‐positive samples was 6.5% and ranged from 0% to 86.7%. Of the 65 flocks studied, 25 (38.5%) had at least one Salmonella‐positive sample. Accounting for confounding variability among the hatcheries and broiler companies, we tested whether the probability of detecting Salmonella in GI tracts of the chicks delivered was associated with certain characteristics of parent breeder flocks; hatchery production volume; hatchery ventilation system; hatchery egg‐room conditions; egg incubation, candling, hatching, eggshell and bird separation, and bird‐processing procedures; management of hatchery‐to‐farm transportation; day of week of hatch; weather conditions during transportation; or season of the hatch. Two risk factor models were adopted. The first model indicated that a greater number of parent flocks, manual separation of eggshell and bird, and a greater amount of fluff and feces on tray liners used during hatchery‐to‐farm transportation at delivery were associated with increased probability of detecting Salmonella in chick GI tracts, whereas a greater number of birds in the delivery vehicle was associated with decreased probability. The second model indicated that broiler flocks hatched on Tuesdays versus either Mondays or Thursdays (with no hatches on Wednesdays, Fridays or week‐ends), increased average hatchability of the eggs from the parent flocks, and greater amounts of fluff and feces on the transport tray liners at delivery were all associated with increased probability of detecting Salmonella in chick GI tracts. The results of this study suggest potential management decisions to lessen Salmonella contamination of broilers supplied by commercial hatcheries and areas for further research.     

Comparison of culturing Mycoplasma gallisepticum from fresh eggs and 18-day-old embryos

Twenty-four 70-week-old and sixteen 27-week-old white leghorn hens were challenged with R strain Mycoplasma gallisepticum (MG) by injection into the caudal thoracic air sac and infraorbital sinus. Eggs were collected daily and cultured within 7 days or incubated for 18 days. Vitelline membranes of eggs were cultured directly; in 18-day-old embryos, cultures were taken from the yolk sac, air sacs, and oral cavity. Culture of vitelline membrane of eggs within 2 days was compared with culture of eggs stored 10 days post oviposition. The first MG-positive egg was laid 2 days postinfection (PI). Hens continued to lay positive eggs to the end of the experiments. There was no significant difference in MG recovery between eggs cultured within 2 days and those cultured 10 days post oviposition. MG was isolated at a significantly higher rate from eggs than from 18-day-old embryos. MG was isolated at a higher rate from the yolk sac of 18-day-old embryos than from the air sacs or oral cavity of the same embryos.     

In ovo feeding of creatine pyruvate modulates growth performance,energy reserves and mRNA expression levels of gluconeogenesis and glycogenesis enzymes in liver of embryos and neonatal broilers

The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the growth performance, energy reserves and mRNA expression levels of gluconeogenesis and glycogenesis enzymes in liver of late‐term embryos and neonatal broilers were investigated. After candling on 16 day of incubation, a total of 960 eggs were randomly assigned to three treatments: (i) non‐injected control, (ii) saline group injected with 0.6 ml of 0.75% physiological saline and (iii) Creatine pyruvate group injected with 0.6 ml of physiological saline containing 12 mg CrPyr/egg. After hatching, 120 male chicks with average body weight (BW) were randomly allocated into each treatment group for a 7‐day feeding trial. The results showed that broilers subjected to CrPyr treatment had higher BW than those of the control and saline groups on 1, 3 and 7 day post‐hatch, as well as the yolk sac weight on 19 day of incubation (19 E), the day of hatch and 3 day post‐hatch (p < .05). Compared with the control and saline groups, IOF of CrPyr increased the plasma creatine concentration on the day of hatch, and the plasma pyruvate concentration on the day of hatch and 3 day post‐hatch (p < .05). Moreover, IOF of CrPyr increased the liver pyruvate and glucose concentrations on 19 E and the day of hatch, and the liver glycogen concentration during the experiment (p < .05). Broilers in the CrPyr group showed increased mRNA expression levels of pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen synthase 2 (GYS2) on 19 E and the day of hatch (p < .05). These results indicated that IOF of CrPyr increased energy reserves in liver of embryos and neonatal broilers possibly through upregulating the mRNA expression levels of PC, PEPCK and GYS2, which could benefit the increase of BW in broilers on 7 day post‐hatch.     

In ovo injection of flavanone on bone quality characteristics,biochemical parameters and antioxidant enzyme status of blood in daily chicks

In ovo injection (IOI) of Naringin (N), flavanone was examined on post‐hatch blood biochemical parameters, antioxidant status and bone characteristics. Fertile eggs (n = 700) were distributed in seven groups with 100 eggs. On 14th and 17.5th days of incubation, four groups were injected using 15 or 30 mg active ingredient levels of naringin/0.5 ml saline/egg, low and high level, into amnion sac. Controls include sham (injected normal saline, 0.5 ml/egg on day 14 and 17.5th) and un‐injected group. IOI of high naringin and saline on 14th day of incubation resulted in lower hatchability and then higher mortality in last week of embryonic life. On day hatch, high levels of injected groups more body weight compared to the control. Chick length was increased at high levels of naringin on day 17.5th compared to control and saline injected. Quality traits of bones were improved in naringin‐injected groups compared to control. IOI of naringin influenced thyroid hormones on 14th day of incubation. Naringin groups influenced the Alkaline phosphatase (ALP), Calcium (Ca), superoxide dismutase (SOD), blood biochemical and lipids. Totally, amniotic IOI of naringin in last days of developing embryo may be useful for hatched chick, development of leg long bone or effect on biochemical metabolites by levels of flavanone that it needs more research.     

Evaluation of epididymis storage temperature and cryopreservation conditions for improved mitochondrial membrane potential,membrane integrity,sperm motility and in vitro fertilization in bovine epididymal sperm

The maintaining of the epididymis at lower temperatures during storage and transport improves sperm quality. Our study aimed to test whether epididymis storage temperature (post‐mortem) and sperm cryopreservation affect sperm kinetics, membrane integrity, mitochondrial potential and fertility capacity. Thirty‐six epididymides were collected from 18 bulls after slaughter and divided into two groups: at 4 or 34°C for 2–3 hr. The sperm was collected from the epididymis cauda. The evaluation consisted of computer‐assisted sperm analysis (CASA), SYBR14/PI/JC1 to evaluate membrane integrity, mitochondrial membrane potential (MMP) and measurement of lipid peroxidation (TBARS). The sperm was then frozen using an automatic device. After thawing, sperm samples were evaluated by the same variables and further in vitro fertilization rates. Cryopreservation negatively affected sperm motility in samples stored at 4 and 34°C. Nevertheless, the 4°C samples yielded higher rates of blastocyst formation. Pre‐freeze sperm motility, progressive motility and velocity were higher in sperm from epididymis stored at 4°C while post‐thaw sperm motility, progressive motility and velocity remained the same among samples from epididymis stored at 4 or 34°C. However, with regard to the kinetic patterns, samples collected from epididymis stored at 34°C had lower values when compared to those stored at 4°C prior the cryopreservation process. Our results indicate that epididymis handling conditions after cryopreservation may affect sperm quality after thawing, especially due to compromised MMP in sperm collected from epididymis stored at higher temperatures.     

Effects of storage time on hatchability of artificially incubated Greater Rhea (Rhea americana) eggs

(1) A study was conducted to determine the effects of the length of the storage period on the hatchability of artificially incubated Greater Rhea eggs. Hatchability was evaluated in eggs gathered daily from a captive population and in eggs collected less frequently from a semi-captive population. (2) Eggs form both sites were either immediately incubated after being collected or were stored for 1 to 9 d prior to incubation. (3) The maximum number of days for which an egg could be stored without depressing hatchability (with respect to non-stored eggs) was longer in the eggs collected daily. (4) Eggs collected daily and stored for 4d or more showed total hatchability (28%) and fertile hatchability (43%) which was approximately 30% lower than non-stored eggs or eggs stored for 3 d or less. In the semi-captive population, the total and fertile hatchability of non-stored eggs and of eggs stored for one day were 40% greater than of eggs stored for 2 to 9 d (20 and 34%, respectively). (5) The period for which Greater Rhea eggs could be stored without depressing hatchability varied depending on the frequency of egg collection: non-daily egg collection reduces the possible period of storage.     

朱鹮引种繁殖试验研究

为了建立不同地区朱种群和加快繁殖 ,2 0 0 2年从洋县引种朱 6 0只到秦岭北麓 ,用人工孵化育雏和亲鸟自然孵化育雏相结合的方法 ,并加强幼鸟管理 ,对其中 14对繁殖朱进行了 2年观察研究。结果显示当年产卵 45枚 ,受精卵 35枚 ,受精率 77.78%,出雏 32只 ,成活2 6只 ,成活率 81.2 5%(2 6 / 32 )。 2 0 0 3年产卵 46枚 ,受精卵 40枚 ,受精率 86 .95%,出雏 35只 ,成活 34只 ,成活率 97.1%(34 / 35)。其中 ,2 0 0 2 ,2 0 0 3年亲鸟自然孵化 ,受精率分别为88.89%和 10 0 %,成活率均为 10 0 %,亲鸟自然孵化育雏优于人工孵化育雏