Bile immunoglobulin of the duck (Anas platyrhynchos). I. Preliminary characterization and ontogeny

Immunoglobulin (Ig) of a single class was found in duck (Anas platyrhynchos) bile. Its molecular weight was 890,000; serum IgM was 800,000. Heavy chains were 75,000 for bile Ig, 86,000 for IgM, 67,000 for 7.8S IgG and 37,000 for 5.7S IgG. Antigenic comparison showed that bile Ig resembled IgM but carried additional determinants. The ontogeny of bile Ig was distinct from that of serum IgM and IgG. Thus, duck bile Ig appears to be an IgM-like molecule secreted independently of serum Ig.     

Isolation of influenza virus from wild ducks (Anas platyrhynchos).

In the course of studies on influenza virus ecology, Influenzavirus A was isolated from a cloacal smear from a wild duck (Anas platyrhynchos) caught in west Slovakia. The strain was identified as A(Hav7Nav1). The results of virus isolation experiments from other species of aquatic and other small birds were negative.     

Bilateral seminoma in a duck (Anas platyrhynchos).

A case of bilateral seminoma in an aged Mallard duck (Anas platyrhynchos) is described. Both testes were enlarged. Historically, the testes revealed large irregular seminiferous tubules containing diffuse sheets of large round, hyperchromatic neoplastic spermatogenic cells. Mitotic figures were seen. The growth had spread diffusely in both testes with no normal tissue left. Occurrence of bilateral seminoma without metastasis to other organs is unusual.     

Unilateral seminoma with multiple visceral metastases in a duck (Anas platyrhynchos).

A case of unilateral seminoma with visceral metastases in a Mallard duck (Anas platyrhynchos) is reported. The right testis was markedly enlarged. The liver surface showed multifocal to coalescent regular circular umbilicated greyish-white spots. In addition, multiple rough whitish nodules were evident on the pancreas and the visceral peritoneum lining the intestine. Histologically, the right testicular parenchyma was diffusely affected and replaced by neoplastic growth, consisting of sheets of large round to polyhedral cells with conspicuous vesicular nuclei having distinctly granular chromatin and prominent nucleoli. Sheets of cells with similar features were observed also in the other affected organs. Multiple lung metastases were detected on histology. This is the first known report of seminoma with hepatic, pancreatic, pulmonary and peritoneal metastases in a Mallard duck.     

The innervation of the ureter in the duck (Anas platyrhynchos). A morphological and quantitative study

The morphology and distribution of the innervation in the duck ureter were studied using AChE histochemistry and PGP 9.5 immunohistochemistry. The density of AChE positive ganglia and neurons was calculated in the adventitial and muscular layers both in young and adult birds. Moreover, in order to investigate regional differences in neuronal density, separate neuron counts and neuron density calculations were performed for the upper, intermediate and lower parts of the ureter, and the data were statistically evaluated. Three nerve plexuses located in the tunica adventitia, in the tunica muscularis and in the lamina propria respectively, were observed. Both in young and adult ducks, the density of adventitial neurons was significantly greater in the lower tract than in the upper and intermediate tracts. The findings of the present study suggest that, in birds, the innervation may play a role in ureteric functions such as epithelial mucosecretion, muscular motility, and closure and/or opening of the ureteric papilla.     

Susceptibility of the domestic duck (Anas platyrhynchos) to experimental infection with Toxoplasma gondii oocysts.

A total of 28 domestic ducks were divided into seven groups of four ducks. Six groups were inoculated per os with 10(1), 10(2), 10(3), 10(4), 10(5) and 10(5.7) oocysts Toxoplasma gondii oocysts (K21 strain, which is avirulent for mice), and the remaining group was used as a control. Antibodies to T. gondii were detected in all ducks by the indirect fluorescence antibody test first on day 7 post-inoculation (p.i.). Antibody titres were found in the range of 1:20 to 1:640 depending on the infectious dose of the oocysts. From day 14 p.i. antibody titres increased to 1:80 to 1:20 480. Between days 14 and 28 p.i. (end of the experiment), antibody titres decreased in 14 ducks, remained the same in seven ducks, and continued to increase in three ducks. Bioassay in mice revealed T. gondii in the breast and leg muscles and the heart (100%, n=47), brain (91%, n=22), liver (54%, n=13) and stomach (46%, n=24). The infected ducks showed no clinical signs; however, the results of bioassay indicate that, compared with some gallinaceous birds, domestic ducks were relatively susceptible to T. gondii infection.     

Histology and ultrastructure of the chorioallantoic membrane of the mallard duck (Anas platyrhynchos)

The histology and fine structure of the chorioallantoic membrane of the mallard duck (Anas platyrhynchos), and the density of vessels per millimeter of membrane were assessed between days 12 and 24 of incubation. Light and transmission electron microscopy of the chorioallantoic membrane of the mallard duck after various days of incubation was carried out. Blood vessels within the mesoderm were counted per millimeter of membrane by light microscopy (40x). The chorioallantoic membrane had three distinct layers from day 12 to 24 of incubation, the chorionic epithelium, the mesoderm, and the allantoic epithelium. After day 12, chorionic epithelium consisted of two layers of flattened, elongated epithelial cells interfaced by numerous desmosomes, and separated from the underlying mesoderm by a basement membrane. At this stage, the allantoic epithelium consisted of a single layer of flattened, overlapping cells. Blood capillaries were observed in the mesoderm close to the chorionic epithelium on days 12 and 13; by day 14, these capillaries were located within the chorionic epithelium, forming a capillary sinus. Between days 14 and 16, the chorion underwent cellular and cytological differentiation into three cell types: capillary covering cells, villus cavity cells, and less differentiated basal cells. The mesoderm was composed of a loose matrix of mesenchymal cells and collagen fibrils through which coursed blood and lymphatic vessels. The vascular density in the mesoderm increased rapidly from 4.2+/-0.6 vessels per mm (n = 12) on day 12 to a maximum of 9.4+/-0.3 vessels per mm (n = 15) by day 16. From day 16, the allantoic epithelium had two to three layers of elongated and overlapping cells. The luminal layer of allantoic epithelial cells had microvillus projections and varying numbers of membrane-bound dense vesicles at all stages from day 12 onward. The histologic and ultrastructural features of mallard duck chorioallantoic membrane from day 12 to 24 of incubation were very similar to those described in the chorioallantoic membrane of the chicken (Gallus gallus) from day 8 to 20 of incubation. Much of the information available concerning the CAM of the chicken also may apply to the CAM of the mallard, with timing adjusted to match the developmental time-frame recorded here.     

Topography and neurochemistry of the enteric ganglia in the proventriculus of the duck (Anas platyrhynchos)

The topographical distribution of the enteric ganglia has been investigated in the proventriculus of the duck using protein gene product 9.5 (PGP 9.5) immunohistochemistry. Myenteric ganglia were usually located between the outer longitudinal and the inner circular muscle layer. Submucous ganglia were sparsely distributed and seemed to be substituted by ganglia located in the tunica mucosa. The neurochemical profile of proventricular ganglion cells was also investigated using nicotinamide adenine dinucleotide phosphate reduced-diaphorase (NADPH-d)-histochemistry and pituitary adenylate cyclase activating peptide (PACAP)/galanin (Gal) double-labelling immunohistochemistry. The majority of mucosal ganglion cells were shown to contain the NADPH-d enzyme and both the investigated peptides. These findings provide evidence for the presence of a mucosal ganglionated plexus in the glandular stomach of birds. Moreover, the neurochemical characteristics of this plexus suggest that it plays an important role in regulating several mucosal functions and, in particular, the production and the composition of the gastric juice.     

3 beta-Hydroxysteroid-dehydrogenases of the testis and epididymis of the Peking duck (Anas platyrhynchos L.)

This study is concerned with the histochemical activity of hydrosteroid dehydrogenases (HSD), glucose-6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) in testis and epididymis of 20 male Pekin ducks. Various 3 beta-HSD, the G6PDH, and the 6PGDH were localized in these organs. 3 beta-etiocholane was much quicker utilized by the 3 beta-HSD than epiandrosterone, dehydroepiandrosterone, and androstendiol. The capability of testis Leydig cells of utilizing the 3 beta-etiocholane was higher in winter than in summer. The G6PDH and the 6PGDH in the Leydig cells and basal cell layer of the tubuli seminiferi contorti were active throughout the year. There was weak 3 beta-HSD activity in the epithelia of the ductuli efferentes distales and in the ductus epididymidis from March to June. The G6PDH and the 6PGDH showed positive reaction in the rete testis and ductuli efferentes proximales throughout the year. The 3 beta-HSD activity in testis and epididymis is saisonally different: In summer, there is no 3 beta-HSD activity in both organs. In winter, the 3 beta-HSD is demonstrable in the testis but not in the epididymis. In spring, there is 3 beta-HSD activity in the epididymis but not in the testis.     

Expression of pituitary adenylate cyclase-activating polypeptide in the primary lymphoid organs of the duck Anas platyrhynchos

The expression of pituitary adenylate cyclase-activating polypeptide (PACAP) was studied in the thymus and bursa of Fabricius of the duck Anas platyrhynchos, at different ages, using immunohistochemistry, Western blotting, RT-PCR and sequencing. In the thymus, PACAP immunoreactivity (-ir) was found in lymphoid cells. CD68/ and PGP 9.5/PACAP38 double labelling showed that PACAP was not expressed either in macrophages or in epithelial cells, suggesting that the PACAP-positive cells observed were lymphoid cells. Immunoreactive lymphocytes were observed in the interlobular septa. They increased in number with ageing. In the bursa, PACAP-ir was found in nerve fibres and in a few lymphoid cells. RT-PCR revealed PACAP mRNA expression in the thymus but not in the bursa. These results suggest that PACAP plays a role in the functions of the immune system in birds.     

Cranial joint histology in the mallard duck (Anas platyrhynchos): new insights on avian cranial kinesis

The evolution of avian cranial kinesis is a phenomenon in part responsible for the remarkable diversity of avian feeding adaptations observable today. Although osteological, developmental and behavioral features of the feeding system are frequently studied, comparatively little is known about cranial joint skeletal tissue composition and morphology from a microscopic perspective. These data are key to understanding the developmental, biomechanical and evolutionary underpinnings of kinesis. Therefore, here we investigated joint microstructure in juvenile and adult mallard ducks (Anas platyrhynchos; Anseriformes). Ducks belong to a diverse clade of galloanseriform birds, have derived adaptations for herbivory and kinesis, and are model organisms in developmental biology. Thus, new insights into their cranial functional morphology will refine our understanding of avian cranial evolution. A total of five specimens (two ducklings and three adults) were histologically sampled, and two additional specimens (a duckling and an adult) were subjected to micro‐computed tomographic scanning. Five intracranial joints were sampled: the jaw joint (quadrate‐articular); otic joint (quadrate‐squamosal); palatobasal joint (parasphenoid‐pterygoid); the mandibular symphysis (dentary‐dentary); and the craniofacial hinge (a complex flexion zone involving four different pairs of skeletal elements). In both the ducklings and adults, the jaw, otic and palatobasal joints are all synovial, with a synovial cavity and articular cartilage on each surface (i.e. bichondral joints) ensheathed in a fibrous capsule. The craniofacial hinge begins as an ensemble of patent sutures in the duckling, but in the adult it becomes more complex: laterally it is synovial; whereas medially, it is synostosed by a bridge of chondroid bone. We hypothesize that it is chondroid bone that provides some of the flexible properties of this joint. The heavily innervated mandibular symphysis is already fused in the ducklings and remains as such in the adult. The results of this study will serve as reference for documenting avian cranial kinesis from a microanatomical perspective. The formation of: (i) secondary articular cartilage on the membrane bones of extant birds; and (ii) their unique ability to form movable synovial joints within two or more membrane bones (i.e. within their dermatocranium) might have played a role in the origin and evolution of modern avian cranial kinesis during dinosaur evolution.     

Cell-mediated immune response to influenza virus infections in mice.

The local and systemic cell-mediated immune (CMI) responses to influenza virus infection in mice were examined by leukocyte migration inhibition and lymphocyte-mediated cytotoxicity tests. Mice were inoculated intranasally with 5 50% lethal doses of the A/WSN (H0N1) strain of influenza virus. Cells from the lymph nodes draining the upper and lower respiratory tract were used to measure the local response, and the spleen was the source of cells used for systemic determinations. The local response by pulmonary lymph node cells was greater and appeared earlier than was observed systemically in the spleen. The specificity of the CMI response was investigated by using a heterologous virus strain, A/Jap (H2N2), and recombinants A/Jap-NWS (H2N1) and A/NWS-Jap (H0N1), obtained from a cross between A/Jap (H2N2) and a virus, A/NWS (H0N1), with surface antigenic specificity similar to that of the inoculated virus. From the results of both tests used as correlates of CMI, it appeared that the response was specific against the hemagglutinin component of the inoculated virus. No reactivity was observed against the heterologous virus A/Jap (H2N2) nor against the recombinant A/Jap-NWS (H2N1) bearing the same neuradminidase as that of the inoculated virus.     

Avian paramyxoviruses and influenza viruses isolated from mallard ducks (Anas platyrhynchos) in New Zealand

Summary.  A comprehensive study using virological and serological approaches was carried out to determine the status of live healthy mallard ducks (Anas platyrhynchos) in New Zealand for infections with avian paramyxoviruses (APMV) and influenza viruses (AIV). Thirty-three viruses isolated from 321 tracheal and cloacal swabs were characterized as: 6 AIV (two H5N2 and four H4N6), 10 APMV-1 and 17 APMV-4. Of 335 sera samples tested for AIV antibodies, 109 (32.5%) sera were positive by nucleoprotein-blocking ELISA (NP-B-ELISA). Serum samples (315) were examined for antibody to APMV-1, -2, -3, -4, -6, -7, -8, -9 by the haemagglutination inhibition test. The largest number of reactions, with titres up to ≥1/64, was to APMV-1 (93.1%), followed by APMV-6 (85.1%), APMV-8 (56%), APMV-4 (51.7%), APMV-7 (47%), APMV-9 (15.9%), APMV-2 (13.3%) and APMV-3 (6.0%). All of the H5N2 isolates of AIV and the APMV-1 isolates from this and earlier New Zealand studies had low pathogenicity indices assessed by the Intravenous Pathogenicity Index (IVPI) with the result 0.00 and Intracerebral Pathogenicity Index (ICPI) with results 0.00–0.16. Partial genomic and antigenic analyses were also consistent with the isolates being non-pathogenic. Phylogenetic analysis of the 10 APMV-1 isolates showed 9 to be most similar to the reference APMV-1 strain D26/76 originally isolated in Japan and also to the Que/66 strain, which was isolated in Australia. The other isolate was very similar to a virus (MC 110/77) obtained from a shelduck in France. Received October 29, 2001; accepted February 26, 2002 Published online May 24, 2002     

Evaluation of two different swab transport systems in the detection of avian influenza virus excretion from infected Pekin ducks (Anas platyrhynchos)

The role of wild birds in the epidemiology and ecology of influenza A viruses has long been recognised (Alexander, 2007a). As a result of the emergence of a H5N1 highly pathogenic avian influenza (HPAI) virus and the apparent role of wild birds in its spread across Asia, Europe and Africa, avian influenza (AI) wild bird surveillance has been implemented in many countries including, since February 2006, a mandatory programme in the European Union (CEC, 2006a). In the present study the detection of virus excreted from Pekin ducks (Anas platyrhynchos) infected experimentally with A/mallard/England/2126/07 (H3N6) was investigated over a fourteen day period post-infection using cloacal and oropharyngeal swabs, with (wet) and without (dry) viral transport medium which were collected from each duck in alternating order. For influenza A virus matrix gene RNA detection, wet oropharyngeal swabs were significantly more sensitive than dry oropharyngeal on days 4-5 after infection. For cloacal samples, dry swabs were equivalent or superior to wet swabs throughout the study. Although differences in detection between dry and wet swabs were observed, the qualitative bird-level results were unaffected, meaning that the infection status of individual birds was correctly determined.