Myostatin in black Muscovy duck (Cairina moschata): full-length cDNA cloning and age-dependent mRNA expression compared with IGF-I

1. Insulin-like growth factor-I (IGF-I) and myostatin (MSTN) are a pair of critical positive and negative growth regulators. The aim of the current study was to examine the age-dependent and muscle-specific expression of IGF-I and MSTN mRNAs in black Muscovy ducks in order to understand their roles in regulating the postnatal muscle growth of domestic ducks.

2. The full-length cDNA of the black Muscovy duck MSTN gene was cloned and the age-dependent mRNA expression profile was compared with that of the IGF-I mRNA in skeletal muscles.

3. The cDNA sequence of the MSTN gene was 1128 bp in length and encodes 375 amino acids, with more than 94.9% homology with poultry MSTN genes, and 83.0–92.0% homology with that of human and mammals (accession: KR006339.1).

4. The IGF-I and MSTN mRNA expression exhibited opposite trends in age-dependency and in different muscles: IGF-I mRNA level was high in the early postnatal stage and low in the late mature stage, corresponding positively to growth; while the MSTN mRNA was low in the early stage, increased gradually and reached the highest level in mature muscles, and was negatively related to muscle growth. In the breast muscles, IGF-I mRNA was much higher than in the leg muscles; the opposite effect was seen in MSTN mRNA.

5. These data suggest that the relative expression levels of IGF-I and MSTN are essential determinants in the temporal and muscle-specific regulation of postnatal skeletal muscle growth in Muscovy duck and possibly in other poultry species as well.

     

Cloning and tissue expression profiling of the GnRHR gene of the Muscovy duck (Cairina moschata): polymorphism and association with egg-laying performance

1. cDNA sequence of gonadotropin-releasing-hormone receptor (GnRHR) gene was cloned and an association analysis between mutations and laying performance was conducted.

2. A 1680-bp cDNA sequence of Muscovy duck GnRHR, which encodes 415 amino acids, was obtained and characterised. Phylogenetic analysis revealed that Muscovy duck GnRHR has a close relationship with Gallus gallus GnRHR.

3. There were significantly different expression profiles between 4 age periods in the hypothalamus, pituitary, and ovary. The expression of GnRHR at the age of 36 weeks (laying period) was higher than other time points in the three tissues. GnRHR was expressed in 12 different tissues. The highest expression levels were observed in hypothalamus, pituitary and gonads.

4. A single nucleotide polymorphism detected in the second intron was associated with egg-laying performance. Individuals with genotype TT had better egg-laying performance from individuals with genotypes CC or TC. Therefore, GnRHR could be used as a marker gene for laying performance in Muscovy duck.     

An outbreak of duck viral enteritis (duck plague) in domestic Muscovy ducks (Cairina moschata domesticus) in Illinois.

Duck viral enteritis (DVE) was diagnosed in an outbreak of the disease in a resident population of Muscovy ducks (Cairina moschata domesticus) on a privately owned multispecies game bird production facility in Illinois, where it claimed 625 ducks. This disease condition had not been reported previously in domestic ducks in Illinois. Although other varieties and age groups of domestic waterfowl (i.e., black ducks, rhumen ducks, Pekin ducks, ducklings, and geese) were present on the game bird farm, the morbidity and mortality (100%) in this epornitic was solely limited to adult ducks of the Muscovy lineage. The clinical signs in the affected ducks were lethargy, diarrhea, dehydration, and death within 2-3 hr of onset of symptoms. Gross pathologic changes were nonspecific and included ecchymotic hemorrhage, effusion of fluid and blood within body cavities reflective of an acute systemic infectious disease. Light microscopic findings were necrosis of primarily digestive lining epithelium and variable lymphohistiocytic infiltration within mucosal and serosal connective tissues. Intranuclear inclusions resembling characteristic herpetic (i.e., Cowdry type A) inclusions were observed primarily in the digestive, respiratory, and reproductive tracts; liver; and spleen. Esophageal candidiasis, bacteriosis, and systemic Pasteurella anatipestifer infections, thought to be concurrent or opportunistic infections, were present in several ducks. DVE virus was demonstrated in infected Muscovy duck embryo fibroblast cells by direct DVE virus-specific fluorescent antibody staining.     

Pharmacokinetics and bioavailability of valnemulin in Muscovy ducks (Cairina moschata)

1. A pharmacokinetic study of valnemulin was conducted in healthy Muscovy ducks after intravenous (IV), intramuscular (IM) and oral administrations at a dose rate of 15?mg/kg body weight.

2. Drug concentrations in plasma were determined by high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Pharmacokinetics parameters of valnemulin were analysed by compartmental analysis using the WinNonlin program.

3. After IV administration, valnemulin was widely distributed with a volume of distribution based on a terminal phase (V_z) of 8·19?±?3·07?l/kg, a mean elimination half-life (t_1/2Ke) of 2·63?h, and a clearance (Cl) value of 5·56?±?1·53?l/kg/h. Following intramuscular and oral administration, valnemulin was rapidly absorbed; the C_max was 0·44?±?0·13 and 0·12?±?0·02?µg/ml (achieved at 0·28 and 1·80?h), the t_1/2Ke was 3·17?±?3·83 and 4·83?±?1·81?h, and the absolute bioavailability (F) was 72% and 37%, respectively.

4. The plasma profile of valnemulin exhibited favourable pharmacokinetic characteristics in Muscovy ducks, such as wide distribution, and rapid absorption and elimination, though oral bioavailability was low.     

Food restriction and development of thermoregulation in Muscovy ducklings (Cairina moschata)

1. The interaction between the effects of food restriction and cold stress on the development of body temperature, homeothermy index, metabolic rate and body weight were studied in Muscovy ducklings from hatching to 21 d of age.

2. The control group (ad libitum fed) and the food restricted group (fed to zero growth rate for 9 d) both became homeothermic when they were 2 d old with moderate (10°C) cold stress. At severe cold stress (0°C) the control group was homeothermic 5 days after hatching. However the food restricted group did not reach homeothermy at 0°C and showed a large decrease (to hatching level) in homeothermy index at 10°C and 0°C after 9 d of food restriction.

3. Body temperature was lower in the food‐restricted group during restriction and increased by 1°C after 24 h of ad libitum feeding. During food restriction, resting metabolic rate did not increase with age and was lower than the basal and existence metabolic rate predicted by Aschoff and Pohl (1970) and Kendeigh (1970) respectively.

4. The ratio of metabolisable energy (ME) intake to resting metabolic rate was 3 times lower in the food‐restricted group than in the control group (0.09 and 0.27 respectively) on day 9. The availability of ME was more important than age for the development of thermoregulation in Muscovy ducklings. It is concluded that small improvements in the feeding regimen of young ducklings enhance the endurance and consequently reduce mortality from to environmental cold stress in a scavenger poultry system.     

Comparative study of ovomucoid isolated from Muscovy duck (Cairina moschata), domestic goose (Anser anser) and domestic duck (Anas platyrhynchos)

1. Ovomucoids were purified from Muscovy duck, domestic duck and domestic goose.

2. Peptide maps of cyanogen bromide‐cleaved ovomucoids from Muscovy duck and domestic duck were very similar to one another, but differed from that of goose.

3. Muscovy duck ovomucoid showed the same protease inhibitory pattern as ovomucoid from domestic duck, inhibiting trypsin in the molar ratio of 1:2 and chymotrypsin 1:1.

4. Inhibitory complexes could be detected between chymotrypsin and ovomucoid from both Muscovy and domestic duck, but not from goose, by using non‐denaturing gels.

5. No complexes could be detected between DFP‐inactivated chymotrypsin and any of the ovomucoids.

6. The results show that of ovomucoid from Muscovy duck more closely resembles that from domestic duck than goose.     

Reproductive season and male effect on quantitative and qualitative traits of individually collected Muscovy duck (Cairina moschata) semen

Males of Muscovy duck (Cairina moschata) are mainly used for mule duck production via artificial insemination of females originated from wild mallard duck (Anas platyrhynchos); therefore, the quantity and quality of drake semen play a crucial role. The assessment results of male reaction to sexual stimulation by dummy female and basic semen characteristic (ejaculate volume, sperm concentration and morphology) of 12 individually kept Muscovy drakes carried out during the entire reproductive season are described. The male and period of the reproductive season effect on scored semen traits are documented. In total, 792 individual semen collections and evaluations were performed. The average of positive reaction in the entire reproductive season varied from 90.6% in December and April to 50.0% in July, while for individual males, it varied between 97.1% and 29.0%. Throughout the season, the ejaculate volumes ranged from 0.05 to 2.45 ml, sperm concentration from 0.15 × 10⁹ ml⁻¹ to 4.44 × 10⁹ ml⁻¹, total number of live spermatozoa from 68.0% to 100% and live normal (properly formed, with any deformations) from 51.0% to 99.0%. Our study indicates the necessity of male breeders pre-selection before the onset of the reproductive season, and the need to leave an appropriate number of males to ensure adequate amount of semen for female insemination, especially when using Muscovy drakes (Cairina moschata) for interspecies crossing with Anas platyrhynchos ducks.     

Pharmacokinetics of orally administered tramadol in Muscovy ducks (Cairina moschata domestica)

This study documents the pharmacokinetics of oral tramadol in Muscovy ducks. Six ducks received a single 30 mg/kg dose of tramadol, orally by stomach tube, with blood collection prior to and up to 24 hr after tramadol administration. Plasma tramadol, and metabolites O‐desmethyltramadol (M1), and N,O‐didesmethyltramadol (M5) concentrations were determined by high‐pressure liquid chromatography (HPLC) with fluorescence (FL) detection. Pharmacokinetic parameters were calculated using a one‐compartment model with first‐order input. No adverse effects were noted after oral administration. All ducks achieved plasma concentrations of tramadol above 0.10 μg/ml and maintained those concentrations for at least 12 hr. Elimination half‐life was 3.95 hr for tramadol in ducks, which is similar to other avian species. All ducks in this study produced the M1 metabolite and maintained plasma concentrations above 0.1 μg/ml for at least 24 hr.     

Cloning and expression analysis of PRL and PRLR genes in black Muscovy duck

ABSTRACT

1. Prolactin hormone (governed by the PRL gene) is secreted by the anterior pituitary of animals, which combines with its receptor (prolactin receptor, PRLR) to act on target cells. Both PRL and PRLR are mainly associated with reproductive performance. The genetic mechanism of nesting in poultry is not yet clear, and so the aim of the current study was to determine expression patterns of PRL and PRLR at different times across the breeding stages of black Muscovy ducks.

2. In this study, the CDS regions of PRL and PRLR were determined by RACE sequencing. The expression levels of PRL and PRLR in the pituitary, ovary and uterus from the black Muscovy duck were compared and analysed during the pre-laying, laying and nesting periods.

3. The results showed that PRL and PRLR are highly homologous in a variety of poultry species. The expression of the PRL gene in the pituitary was the highest, which was significantly higher than seen in the ovary and uterus. This trend ran through the entire prenatal period, i.e. the laying period and the nesting period. The expression level of the PRLR gene in the pituitary and ovary was generally low, and expression in the uterus was the highest. There was no significant difference in expression of the PRLR gene between pituitary and ovary during different periods, but the expression level of the PRLR gene in the uterus reached its highest level during the nesting stage, which was significantly higher than seen in the early laying period.     

Development of prenatal acoustic interaction in the Muscovy duck (Cairina Moschata)

1. The development of prenatal acoustic communication was investigated in Muscovy ducks, comparing the vocalisation activities and call types in isolated eggs and pairs of embryos (two eggs in shell contact).

2. Incubator experiments showed that the number of vocalisations in embryos which had mutual acoustic contact was nearly twice that of isolated ones.

3. An acoustic interaction, that is, an exchange of signals, was only recognised after the first chick had hatched, that is, between embryos and chicks.

4. Specific call types used for communication were the double call and the trill call.     

Comparative pathological studies on domestic geese (Anser anser domestica) and Muscovy ducks (Cairina moschata) experimentally infected with parvovirus strains of goose and Muscovy duck origin

Parvovirus infection of Muscovy ducks caused by a genetically and antigenically distinct virus has been reported from Germany, France, Israel, Hungary, some Asian countries and the USA. The pathological changes include those of degenerative skeletal muscle myopathy and myocarditis, hepatitis, sciatic neuritis and polioencephalomyelitis. In the study presented here, day-old and 3-week-old goslings and Muscovy ducks were infected experimentally with three different parvovirus strains (isolates of D-216/4 from the classical form of Derzsy's disease, D-190/3 from the enteric form of Derzsy's disease, and strain FM from the parvovirus disease of Muscovy ducks). All three parvovirus strains caused severe disease in both day-old and 3-week-old Muscovy ducks but in the goslings only the two strains of goose origin (D-216/4 and D-190/3) caused disease with high (90-100%) mortality when infection was performed at day old. Strain FM (of Muscovy duck origin) did not cause any clinical signs or pathological lesions in the goslings. In the day-old goslings and Muscovy ducks the principal pathological lesions were severe enteritis with necrosis of the epithelial cells (enterocytes) of the mucous membrane and the crypts of Lieberkühn, and the formation of intranuclear inclusion bodies. Other prominent lesions included hepatitis and atrophy (lymphocyte depletion) of the lymphoid organs (bursa of Fabricius, thymus, spleen). In goslings infected with the strain originating from the classical form of Derzsy's disease mild myocarditis was also detected. After infection at three weeks of age, growth retardation, feathering disorders, myocardial lesions (degeneration of cardiac muscle cells, lympho-histiocytic infiltration) and hepatitis were the most prominent lesions in both geese and Muscovy ducks. In addition to the lesions observed in the geese, muscle fibre degeneration, mild sciatic neuritis and polioencephalomyelitis were also observed in the Muscovy ducks infected with any of the three parvovirus strains.     

Photorefractory period of the Muscovy duck (Cairina moschata): endocrine and neuroendocrine responses to day length after a full reproductive cycle

1. The Muscovy duck, of equatorial origin, is photosensitive and its sexual maturation can be advanced by long days. The aim of this study was to investigate the photorefractory nature of the seasonal sexual rest in this species.

2. Sixty males were allocated to three groups of 20 birds each after completion of one full reproductive cycle in long days (16L:8D). Group A was transferred to longer days (20L:4D). Group B remained in 16L:8D. Group C was transferred to short days (6L: 18D) for 9 weeks and then returned to 16L:8D.

3. Group B spontaneously redeveloped their testes and recrudescence, indicated by increased plasma concentrations of luteinising hormone and testosterone, started between the 4th and 10th week after total regression. Longer days (in group A) accelerated recrudescence. Shorter days (in group C) delayed it, but the return to long days restored reproductive function.

4. In group C, the transfer to short days increased the hypothalamic content of luteinising hormone‐releasing hormone (LHRH) and secretion of the neuropeptide in vitro. The pituitary sensitivity to LHRH was increased in that group after the return to long days.

5. These effects show that the birds were photosensitive when daylength was changed 8 weeks after the completion of regression, and that short days were not necessary for the birds to regain photosensitivity.

6. The shortness of the regressed period and the spontaneous recrudescence in long days are more typical of equatorial birds than temperate zone species, which are more dependent on photoperiodic changes. The Muscovy duck probably retains features of its wild ancestor, which, living in equatorial latitudes, was more subject to non‐photic synchronisers.     

Effect of semen extenders and storage time on quality of Muscovy duck (Cairina moschata) drake semen during the entire reproductive season

Avian semen dilution with appropriate extender allows to prolong the fertilizing ability of sperm stored in vitro. In the present study, the impact of extenders and time of storage on morphology of Muscovy duck (Cairina moschata) drake semen were examined. Semen was collected twice a week, using male stimulation by a female method, from 12 adults (29 weeks old) drakes kept individually in cages, under controlled environmental conditions. Freshly collected, pooled ejaculates were divided into three part: neat undiluted sample, and diluted 1:1 with Schramm (SCH) or Watanabe (W) extender and stored at 4°C. Morphological examination of all samples was conducted after dilution and then, after 3 and 6 hr of storage. The storage of undiluted semen caused decrease (p ≤ .01) in live morphologically normal sperm, from 79.73% in the freshly collected ejaculates to 55.75% and to 12.12% after 3 and 6 hr of storage, respectively (average calculated for the entire reproductive season). In the semen diluted with Schramm's extender the adequate values attained 86.84, 79.65 and 61.66%, and using Watanabe extender 84.77, 83.58 and 75.25%, respectively. The period of semen storage and the type of extender caused significant (p ≤ 0,05; p ≤ 0,01) changes in sperm morphology. The longer period of storage contributed to the decrease in number of morphologically normal sperm, whereas their content in Watanabe extender after 3 and 6 hr of storage was higher (p ≤ .01) than in semen diluted in Schramm extender.     

Morphological and glycohistochemical studies on the epididymal region of the Sudani duck (Cairina moschata)

In this study, the epididymal region of the Sudani duck was investigated using histological and lectin histochemical methods. Morphologically, the epididymal region of the Sudani duck is composed of extratesticular rete testis, proximal and distal efferent ductules, a short connecting duct, and epididymal ducts. Morphometric analysis of the epididymal region of Sudani duck revealed that the efferent ductules predominate in relation to the epididymal ducts. The distribution of sugar moieties within the epididymal region of the Sudani duck was investigated using ten different fluorescein isothiocyanate (FITC) conjugated lectins. In the rete testis epithelium, only PHA-L showed a positive reaction. Efferent ductules in contrary exhibited a wide range of lectin affinity whereas six positive lectins (Con A, LCA, PNA, WGA, PHA-L, PHA-E) were observed. In the connecting and epididymal ducts, four lectins (Con A, WGA, PHA-L, PHA-E) were also detected. GSA-I, UEA-I, and LTA were at all not evident in the epididymal region of the Sudani duck. In conclusion, the correlation between the large areas of the epididymal region occupied by the efferent ductules and the wide range of sugar affinity of this portion may confirm the speculation that efferent ductules might be the primary site of fluid reabsorption in the epididymal region of Sudani duck.     

The first outbreak of fowl cholera in Muscovy ducks (Cairina moschata) in Japan.

The first outbreak of fowl cholera occurred in a flock of Muscovy ducks (Cairina moschata) in Okinawa Prefecture of Japan in November 1990. Fifty (25%) of 200 birds in a farm died of an acute disease. Remaining birds recovered after treatment with oxytetracycline. Pasteurella multocida subsp. multocida was isolated in pure culture from all tissues tested from two dead birds. Serovars of the isolates were identified as Carter's capsular type A. Heddleston's type 3.4.12, and Namioka's type 5:A which have not been demonstrated in Japan. Pathologically, multiple necrosis and bacterial aggregates were prominent in several organs, particularly in the liver. The isolate killed chickens when inoculated intravenously at a concentration of 10(8) colony forming units.     

First report of parasitic esophagitis by Streptocara incognita in Muscovy ducks (Cairina moschata domesticus) in Italy

This work describes the first outbreak of streptocariasis by Streptocara incognita reported from Italy. The disease was observed in a backyard flock of 62 ducks (Cairina moschata domesticus) located in Treviso, northeastern Italy. Fifteen birds died in a period of 2 wk, showing debilitation and emaciation. Two ducks were submitted for postmortem examination and an esophagitis associated with nematodes was observed. The nematodes were identified as Streptocara incognita.     

Requirements of Muscovy ducks (Cairina moschata domestica) for crude protein and energy levels of fattening diets

Groups of male and female Cairina ducks subjected to equal starter feeding (1st to 3rd weeks) were used from the 4th week of live (fattening period) to test feed mixes of varying energy and crude protein levels (480 energetic feed equivalents/heu[EFH] - 140 g crude protein[CP]; 480 EFh - 180/g CP; 555 EFh - 140 g CP; 555 EFh - 180 g CP; 625 EEh - 140 g CP; 625 EFh - 180 g CP - all figures referred to 1 kg of feed). The different energy levels remained without influence of the live weight development. High-energy feed caused, however, higher expenditures of energetic feed equivalents per kg of gain and resulted in carcases with higher fat proportion. At all the 3 energy levels under study, high-protein feeding proved superior to low-protein rations with regard to live weight gains and meat yield. For the time being, 500 EFh and 180 g CP per kg of organic matter are recommended as indicators for Cairina duck fattening rations.     

Pharmacokinetics of doxycycline after a single intravenous,oral or intramuscular dose in Muscovy ducks (Cairina moschata)

1. The pharmacokinetics of doxycycline in ducks were investigated after a single intravenous (IV), intramuscular (IM) or oral (PO) dose at 20 mg/kg body weight.

2. The concentrations of doxycycline in plasma samples were assayed using a high performance liquid chromatography method, and pharmacokinetic parameters were calculated using a non-compartmental model.

3. After IV administration, doxycycline had a mean (±SD) distribution volume (V_z) of 1761.9 ± 328.5 ml/kg and was slowly eliminated with a terminal half-life (t_1/2λz) of 21.21±1.47 h and a total body clearance (Cl) of 57.51 ± 9.50 ml/h/kg. Following PO and IM administration, doxycycline was relatively slowly absorbed – the peak concentrations (C_max) were 17.57 ± 4.66 μg/ml at 2 h and 25.01 ± 4.18 μg/ml at 1.5 h, respectively. The absolute bioavailabilities (F) of doxycycline after PO and IM administration were 39.13% and 70.71%, respectively.

4. The plasma profile of doxycycline exhibited favourable pharmacokinetics characteristics in Muscovy ducks, such as wide distribution, relatively slow absorption and slow elimination, though oral bioavailability was low.

     

番鸭白细胞介素2基因的克隆及其原核表达

为原核表达番鸭白细胞介素2 (MdIL-2),本研究利用ConA刺激诱导番鸭脾淋巴细胞,采用RT-PCR技术扩增MdIL-2基因的编码序列,并将其克隆至pGEX-6P-1中,构建重组质粒pGEX-MdIL-2,转化E.coli BL21受体菌.测序结果表明,MdIL-2基因ORF全长420 bp,编码140个氨基酸.重组菌经IPTG诱导表达后,SDS-PAGE分析显示,重组蛋白约为40 ku.Western blot分析表明,重组蛋白能够被抗GST单克隆抗体特异性识别.MdIL-2基因的克隆及其原核表达为进一步进行MdIL-2的活性检测以及应用研究奠定了基础.