Histochemical Analysis of Glycoconjugates in the Skin of a Catfish (Arius Tenuispinis, Day)

A histochemical study using conventional carbohydrate histochemistry (periodic‐acid staining including diastase controls, alcian blue staining at pH 1 and 2.5) as well as using a battery of 14 fluorescein isothiocyanate (FITC)‐labelled lectins to identify glycoconjugates present in 10 different areas of the skin of a catfish (Arius tenuispinis) was carried out. The lectins used were: mannose‐binding lectins (Con A, LCA and PSA), galactose‐binding lectins (PNA, RCA), N‐acetylgalactosamine‐binding lectins (DBA, SBA, SJA and GSL I), N‐acetylglucosamine‐binding lectins (WGA and WGAs), fucose‐binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E, PHA L). Conventional glycoconjugate staining (PAS staining, alcian blue at pH 1 and 2.5) showed that the mucous goblet cells contain a considerable amount of glycoconjugates in all locations of the skin, whereas the other unicellular gland type, the club cells, lacked these glycoconjugates. The glycoproteins found in goblet cells are neutral and therefore stain magenta when subjected to PAS staining. Alcian blue staining indicating acid glycoproteins was distinctly positive at pH 1, but gave only a comparable staining at pH 2.5. The mucus of the goblet cells therefore also contains acid glycoproteins rich in sulphate groups. Using FITC‐labelled lectins, the carbohydrate composition of the glycoproteins of goblet cells could be more fully characterized. A distinct staining of the mucus of goblet cells was found with the mannose‐binding lectins LCA and PSA; the galactosamine‐binding lectins DBA, SBA and GLS I; the glucosamine‐binding lectin WGA; and PHA E which stains glycoproteins with complex carbohydrate configurations. No reaction occurred with the fucose‐binding lectin UEA and the sialic acid‐specific lectin SNA. In addition, the galactose‐binding lectins PNA and RCA showed only a weak or completely negative staining of the mucus in the goblet cells. The specificity of the lectin staining could be proved by inhibiting binding of the lectins by competitive inhibition with the corresponding sugars. From these data, we can conclude that the mucus produced by the epidermal goblet cells of A. tenuispinis is rich in mannose, N‐acetylgalactosamine and N‐acetylglucosamine residues.     

Heterogeneous expression of glycoconjugates in the primary olfactory centre of the Japanese sword‐tailed newt (Cynops ensicauda)

Histochemical organization of the Caudata olfactory system remains largely unknown, despite this amphibian order showing phylogenetic diversity in the development of the vomeronasal organ and its primary centre, the accessory olfactory bulb. Here, we investigated the glycoconjugate distribution in the olfactory bulb of a semi‐aquatic salamander, the Japanese sword‐tailed newt (Cynops ensicauda), by histochemical analysis of the lectins that were present. Eleven lectins showed a specific binding to the olfactory and vomeronasal nerves as well as to the olfactory glomeruli. Among them, succinylated wheat germ agglutinin (s‐WGA), soya bean agglutinin (SBA), Bandeiraea simplicifolia lectin‐I (BSL‐I) and peanut agglutinin showed significantly different bindings to glomeruli between the main and accessory olfactory bulbs. We also found that s‐WGA, SBA, BSL‐I and Pisum sativum agglutinin preferentially bound to a rostral cluster of glomeruli in the main olfactory bulb. This finding suggests the presence of a functional subset of primary projections to the main olfactory system. Our results therefore demonstrated a region‐specific glycoconjugate expression in the olfactory bulb of C. ensicauda, which would be related to a functional segregation of the olfactory system.     

The Accessory Olfactory Bulb of the Mink, Mustela vison: a Morphological and Lectin Histochemical Study

The distribution of binding sites for the lectins Ulex europaeus agglutinin I. Soybean agglutinin, Bandeiraea simplicifolia agglutinin I-isolectin B₄, and Vicia villosa agglutinin in the mink olfactory bulb was investigated. All lectins except Ulex europaeus agglutinin I bound exclusively and systematically to a single area of the olfactory bulb. This area corresponded to that in which the vomeronasal nerves terminate, indicating that it is the accessory olfactory bulb, as confirmed by microdissection and by the study of transverse and parasagittal series of the olfactory bulb. The results, moreover, indicate that the accessory olfactory bulb of the mink comprises three isolated eminences, the largest in the dorsal part of the olfactory bulb, and the other two in the lateral and medial parts.     

Characterization of glycoconjugates and sialic acid modification in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis)

Diverse glycoconjugates are expressed in the vertebrate olfactory bulb and serve as guidance cues for axons of nasal receptor neurons. Although the involvement of glycoconjugates in the segregation of the olfactory pathway has been suggested, it is poorly understood in salamanders. In this study, lectin histochemistry was used to determine glycoconjugate distribution in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis). Succinylated wheat germ agglutinin (sWGA), Ricinus communis agglutinin-I and Lens culinaris agglutinin showed different bindings in the nerve fibre layer or glomerular layer, or both, between the main and accessory olfactory bulbs. We then investigated the lectin-binding pattern after the removal of terminal sialic acids using neuraminidase. Desialylation resulted in a change in the binding reactivities with seven lectins. Wheat germ agglutinin, sWGA, soybean agglutinin (SBA) and peanut agglutinin showed different degrees of binding between the main and accessory olfactory bulbs. In addition, SBA showed a heterogeneous labelling of glomeruli in the rostral region of the main olfactory bulb. Our results suggest that terminal sialic acids mask the heterogeneity of glycoconjugates in the olfactory bulb of C. orientalis.     

Lectin Histochemical Analysis of the Olfactory Bulbs in the Barfin Flounder (Verasper moseri)

Several lines of evidence have shown that the olfactory system of the fish contains the main and accessory olfactory systems. However, morphological data indicate that the accessory olfactory bulb, the primary centre for the accessory olfactory system, will not differentiate in the fish. Therefore, the fish olfactory bulb is supposed to engage in both main and accessory olfactory systems. To examine this possibility, we investigated the olfactory bulb of the barfin flounder ( Verasper moseri ) by histochemical examination using lectins. The olfactory bulb of the barfin flounder showed a laminar structure with four layers, and diffuse glomerular architecture was observed in the glomerular layer. Based on the expression patterns of sugar residues, the glomerular layer of the barfin olfactory bulb was largely divided into three portions. Heterogeneity in the lectin-binding pattern among olfactory glomeruli was clearly demonstrated by the fluorescent double-lectin staining. The results of this study suggest that the fish olfactory bulb contains both regions equivalent to the main and accessory olfactory bulbs, and they are subdivided into small subsets with different functions.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Evaluation of growth performance, serum biochemistry and haematological parameters on broiler birds fed with raw and processed samples of Entada scandens, Canavalia gladiata and Canavalia ensiformis seed meal as an alternative protein source

The experiment was carried out to investigate the inclusion of underutilised legumes, Entada scandens, Canavalia gladiata and Canavalia ensiformis, seed meal in soybean-based diet in broilers. The utilisation of these wild legumes is limited by the presence of antinutrient compounds. Processing methods like soaking followed by autoclaving in sodium bicarbonate solution in E. scandens and C. gladiata and soaking followed by autoclaving in ash solution in C. ensiformis were adopted. The proximate composition of raw and processed samples of E. scandens, C. gladiata and C. ensiformis were determined. The protein content was enhanced in processed sample of E. scandens (46 %) and C. ensiformis (16 %). This processing method had reduced the maximum number of antinutrients such as tannins (10–100 %), trypsin inhibitor activity (99 %), chymotrypsin inhibitor activity (72–100 %), canavanine (60–62 %), amylase inhibitor activity (73–100 %), saponins (78–92 %), phytic acid (19–40 %) and lectins. Hence, the raw samples at 15 % and processed samples at 15 and 30 % were replaced with soybean protein in commercial broiler diet respectively. Birds fed with 30 % processed samples of E. scandens, C. gladiata and C. ensiformis showed significantly similar results of growth performance, carcass characteristics, organ weight, haematological parameters and serum biochemical parameters (cholesterol, protein, bilirubin, albumin, globulin and liver and kidney function parameters) without any adverse effects after 42 days of supplementation. The proper utilisation of these underutilised legumes may act as an alternative protein ingredient in poultry diets.     

Goblet Cell Types in Intestine of Tiger Barb and Black Tetra (Cyprinidae,Characidae: Teleostei)

Histochemical properties of goblet cells in intestine of a stomach‐less teleost, tiger barb (Puntius tetrazona), and a stomach‐containing teleost, black tetra (Gymnocorymbus ternetzi), are described and compared. The intestine goblet cells were mostly wide in both species, but in tiger barb, some of them were markedly thinner. In black tetra, all the intestine goblet cells displayed magenta colour after PAS, whereas in the tiger barb, only the thinner goblet cells displayed such affinity. The latter cell type was coloured strongly magenta when the tissue was treated with alcian blue (pH 2.5) followed by PAS, whereas the wide goblet cells in tiger barb and all goblet cells in black tetra displayed mainly a blue colour after such treatment. Further, the goblet cells in both species were coloured cleanly blue after high iron diamine followed by alcian blue (pH 2.5). The intestine goblet cells in both species displayed a moderate affinity to WGA and concanavalin A lectins and no affinity to DBA. Most of the goblet cells displayed no affinity to PNA, but some of them in the tiger barb displayed a moderate or strong affinity to this lectin. The affinity to WGA was somewhat strengthened after pre‐treatment with neuraminidase. These results suggest that tiger barb contains two types or variants of intestinal goblet cells: high numbers of wide cells filled by acidic, non‐sulphated mucin and some thinner cells filled by neutral mucin. The intestine goblet cells in black tetra were filled by variable amounts of neutral and acidic mucin, but the total number of such cells is much less than in tiger barb. The present lectin and neuraminidase results suggest that the intestinal mucins in both species contain significant amounts of N‐acetylglucosamine, sialic acid and glucose/mannose, but seem to lack N‐acetylgalactosamine. However, some of these cells in tiger barb contain moderate to large amounts of galactose. Together, these results suggest significant species‐specific features of the intestine goblet cells and mucin types in tiger barb and black tetra. In conclusion, the present results suggest that the diet and feeding habits in stomach‐less teleosts compared with stomach‐containing teleosts, greatly influence the number of intestine goblet cells and type of mucin in these cells.     

Pharmacokinetics and pharmacodynamics of alfaxalone after a single intramuscular or intravascular injection in mallard ducks (Anas platyrhynchos)

Pharmacokinetics and pharmacodynamics of alfaxalone was performed in mallard ducks (Anas platyrhynchos) after single bolus injections of 10 mg/kg administered intramuscularly (IM; n = 10) or intravenously (IV; n = 10), in a randomized cross‐over design with a washout period between doses. Mean (±SD) C_max following IM injection was 1.6 (±0.8) µg/ml with T_max at 15.0 (±10.5) min. Area under the curve (AUC) was 84.66 and 104.58 min*mg/ml following IV and IM administration, respectively. Volume of distribution (V_D) after IV dose was 3.0 L/kg. The mean plasma clearance after 10 mg/kg IV was 139.5 (±67.9) ml min^?1 kg^?1. Elimination half‐lives (mean [±SD]) were 15.0 and 16.1 (±3.0) min following IV and IM administration, respectively. Mean bioavailability at 10 mg/kg IM was 108.6%. None of the ducks achieved a sufficient anesthetic depth for invasive procedures, such as surgery, to be performed. Heart and respiratory rates measured after administration remained stable, but many ducks were hyperexcitable during recovery. Based on sedation levels and duration, alfaxalone administered at dosages of 10 mg/kg IV or IM in mallard ducks does not induce clinically acceptable anesthesia.     

Distribution of Lectin‐Bindings in the Testis of the Lesser Mouse Deer,Tragulus javanicus

The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d ‐galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N‐acetyl‐d ‐galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N‐acetyl‐d ‐glucosamine and sialic acid (wheat germ agglutinin WGA, s‐WGA), d ‐mannose and d ‐glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), l ‐fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA‐E) sugar residues. In Golgi‐, cap‐, and acrosome‐phase spermatids, lectin‐bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s‐WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA‐E). s‐WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA‐E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA‐E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA‐E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin‐bindings noted in the testis of lesser mouse deer included the limited distribution of s‐WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.     

Intermediate Filament Proteins Expression and Carbohydrate Moieties in Trophoblast and Decidual Cells of Mature Cat Placenta

The aim of this study was to characterize cytoskeletal intermediate filament proteins and glycoconjugates of syncytiotrophoblast, cytotrophoblast and decidual cells of feline endotheliochorial placenta. Samples from 12 normal pregnant female cats, after 45 ± 5 days of gestation, were obtained removing the uterine horns by hysterectomy. Sections were processed for routine observation and for immunohistochemistry using anticytokeratin, antivimentin and antidesmin antibodies. In addition, lectin histochemistry was performed using a panel of several biotinylated lectins to characterize glycosides expression profile. Cytotrophoblast and syncytiotrophoblast showed immunoreactivity only with acidic and basic cytokeratins. Decidual cells were only positive to vimentin, consistent with their origin from endometrial fibroblasts. Trophoblast expressed a broad population of glycans, highly exposing terminal N‐acetyl glucosamine residues and non‐sialylated galactose and N‐acetyl galactosamine oligomers. Oligosaccharides bound by Phaseolus vulgaris erythroagglutinin were the only highly branched N‐linked residues evidenced in cats, and they were restricted to the syncytium. Unlike results reported on humans, mice and rats on lectin affinity of decidual cells, sialid acids and complex N‐linked oligosaccharides were not demonstrated in cats. Glycosylation of proteins determines many of their final properties, thus becoming essential for the embryo‐maternal dialogue during implantation and placentation. Changes in glycosylation pattern have been related to pathological pregnancies in other species. Hence, the knowledge about glycosylation profile of the normal cat placenta may lead to a better understanding of both normal and pathological reproductive events.     

Lectin Binding Characteristics of the Olfactory Mucosa of Channel Catfish: Potential Factors in Attachment of Edwardsiella ictaluri

Abstract

The olfactory organ is a primary infection site for Edwardsiella ictaluri, the etiologic agent of enteric septicemia of channel catfish Ictalurus punctatus. The olfactory mucosal surface is a major interface between host and pathogen where commonly occurring carbohydrates may act as receptors for bacterial attachment. In this study, d-mannose, N-acetylgalactosamine, N-acetylglucosamine, N-acetylneuraminic acid, d-galactose, and l-fucose were histochemically localized in the olfactory mucosa of channel catfish by using lectins that preferentially bind these carbohydrates. These lectins were Concanavalin A (ConA), soybean agglutinin (SBA), pokeweed agglutinin (PWA), wheat germ agglutinin (WGA), peanut agglutinin (PNA), and Ulex europaeus agglutinin I (UEA-I), respectively. The olfactory mucosa expressed d-mannose ubiquitously, whereas l-fucose and N-acetylneuraminic acid expression was specific to the apical mucosal surface. The carbohydrates d-galactose, N-acetylgalactosamine, and N-acetylglucosamine were most abundant in the sensory mucosa, specifically olfactory receptor neurons and cells near the basal lamina. Edwardsiella ictaluri was assayed for carbohydrate affinities by colloidal gold immunolocalization and transmission electron microscopy. Of the anti-lectins examined, those against WGA and UEA-I cross-reacted most intensely with Edwardsiella ictaluri, whereas cross-reactivities of anti-ConA, -SBA, and -PNA were more moderate. Double immunofluorescence labeling of experimentally infected catfish showed E. ictaluri adherent to cell surfaces or intercellularly associated with labeled carbohydrate components of the olfactory mucosa. Preincubation of the olfactory mucosa with soluble d-galactose significantly reduced bacterial adhesion compared with controls. Our results indicate a specific pattern of carbohydrates present in the catfish olfactory mucosa and suggest carbohydrates participate in initial E. ictaluri attachment by acting as ligands for pathogen constituents.     

Plasma pharmacokinetics of quinocetone in ducks after oral and intravenous administration

Quinocetone (QCT), an antimicrobial growth promoter, is widely used in food‐producing animals. However, information about pharmacokinetics (PK) of QCT in ducks still remains unavailable up to now. In this study, QCT and its major metabolites (1‐desoxyquinocetone, di‐desoxyquinocetone and 3‐methyl‐quinoxaline‐2‐carboxylic) in ducks were studied using a simple and sensitive UHPLC‐MS/MS assay. Twenty ducks were divided into two groups. (n = 10/group). One group received QCT by oral administration at dose of 40 mg/kg while another group received QCT intravenously at 10 mg/kg. Plasma samples were collected at various time points from 0 to 96 hr. QCT and its major metabolites in duck plasma samples were extracted by 1 ml acetonitrile and detected by UHPLC‐MS/MS, with the gradient mobile phase that consisted of 0.1% formic acid in water (A) and acetonitrile (B). A noncompartment analysis was used to calculate the PK parameters. The results showed that following oral dosing, the peak plasma concentration (C_max) of QCT was 32.14 ng/ml and the area under the curve (AUCINF_obs) was 233.63 (h ng)/ ml. Following intravenous dosing, the C_max, AUCINF_obs and Vss_obs were 96.70 ng/ml, 152.34 (h ng)/ ml and 807.00 L/kg, respectively. These data indicated that the QCT was less absorbed in vivo following oral administration, with low bioavailability (38.43%). QCT and its major metabolites such as 1‐desoxyquinocetone and 3‐methyl‐quinoxaline‐2‐carboxylic were detected at individual time points in individual ducks, while the di‐desoxyquinocetone was not detected in all time points in all ducks. This study enriches basic scientific data about pharmacokinetics of QCT in ducks after oral and intravenous administration and will be beneficial for clinical application in ducks.     

Mucus characterisation in the Octopus vulgaris skin throughout its life cycle

The development of the epidermis of octopus, Octopus vulgaris, throughout its life cycle was studied by conventional staining and histochemical techniques using lectins. The mantle, the arm and the two parts of the suckers: the infundibulum and the acetabulum were analysed independently. With the exception of the suckers, the general morphology of the epidermis does not vary from the first days post-hatching to adulthood. In general terms, histochemical techniques do not indicate changes in the composition of glycoconjugates of the epidermis main cells, epithelial and secretory cells. The epithelial cells of the mantle and arm show positivity for mannose (ConA+) in their apical portions, indicating the presence of n-glycoproteins that, among other things, provide lubrication to the surface of the body. In the suckers, the apical surface of the infundibulum contains sulphated glycosaminoglycans of the N-acetylglucosamine type that provide adhesive properties. In addition to observing three types of mucocytes, m1 and m2 are characteristic of the mantle and arm, and m3 is found in the suckers. The paralarva epidermis is characterised by the presence of Kölliker's organs whose exact function is unknown. In this study, the absence of staining with alcian blue/periodic acid–Schiff(AB/PAS) prevents the possibility of attributing a secretory function. Nevertheless, the linkage of three lectins (WGA, LEL and GSL-I) in the fascicle of the organ suggests the presence of proteoglycans rich in N-acetylglucosamine that would mainly have a structural role.     

Topography of ganglion cells in the retina of the duck (Anas platyrhynchos var. domesticus)

The purpose of this study was to define ganglion cell density, size and topography in the retina of the mallard duck. After killing adult mallard ducks (Anas platyrhynchos var. domesticus), their eyes were removed using pentobarbital (30 mg/kg). The retinas were isolated, whole mount specimens were prepared by staining with 0.1% cresyl violet and then fixing the tissues for study. The retinal ganglion cells were counted, mapped and measured. The mean total number of ganglion cells was estimated at approximately 1.7 × 10⁶ and the retinal area centralis had the highest ganglion cell density with 15 820 cells/mm². The number of ganglion cell bodies was highest in the temporal area, followed by the nasal, dorsal and ventral areas. Ganglion cell size ranged from 56 to 406 μm². A population of small ganglion cells persisted into the central area just above the optic disc and the largest soma area was in the ventral zone of the retina. This localization of ganglion cells suggests that the quality of vision is not equal in all the areas of the duck retina and the central part may have the highest vision quality as a function of the retinal ganglion cells.     

Surveillance for highly pathogenic avian influenza in migratory shorebirds at the terminus of the East Asian-Australasian Flyway

AIM: To determine if migratory birds arriving in New Zealand in the Southern Hemisphere spring of 2004 were infected with the highly pathogenic avian influenza (AI) virus, H5N1.

METHODS: Cloacal and faecal samples were collected from migratory red knots following their arrival in New Zealand in October 2004. Two species of resident sympatric birds, wrybill and mallard duck, were sampled prior to, and following, the arrival of migratory birds.

RESULTS: No AI viruses were isolated from migratory or resident shorebirds. Non-pathogenic AI viruses were isolated from six resident mallard ducks, comprising the endemic subtypes H4 (n=2), H7 (non-pathogenic), H10, and H11 (n=2).

CONCLUSIONS: Highly pathogenic AI H5N1 virus was not detected in migratory shorebirds or sympatric water birds in the Firth of Thames, New Zealand, in 2004-2005, despite the possible proximity of migratory birds to outbreaks of the disease in East Asia in 2004.     

Ontogeny of testicular inhibin and activin in ducks: An immunocytochemical analysis

The ontogeny of testicular inhibin/activin in ducks was investigated. Testicular localization of three inhibin/activin subunits (α, β_A and β_B) was determined in embryonic and newly hatched ducks from 12 days of incubation to 1 day of age, in immature ducks and in adult ducks. In the duck embryonic testis, positive α‐subunit immunostaining was first detected in the Leydig cells and Sertoli cells on day 15 of incubation, whereas β_A‐subunit and β_B‐subunit immunostaining were found in Sertoli cells and primary germ cells on day 18 of incubation. In 1 month old ducks, intense staining of α‐subunit was present in the seminiferous epithelium consistent with localization in Sertoli cells and primary germ cells, and the immunostaining of the β_A‐ and β_B‐subunit was also present in Sertoli cells and primary germ cells. Specific immunostaining with inhibin/activin α‐, β_A‐ and β_B‐subunits antisera occurred in Sertoli cells in the adult duck testes. In conclusion, it was shown that, in the duck testis, the majority of α‐, β_A‐ and β_B‐subunits are colocalized in Sertoli cells with a certain degree of staining in germ cells and the α‐subunit is present in Leydig cells of embryonic testes before day 18 of incubation. These results indicate that Sertoli cells and possibly germ cells in the embryonic testes of late stage of incubation and newly hatched ducks, immature ducks and mature ducks may produce bioactive inhibin dimers, inhibin A and inhibin B, as a possible regulator of follicle‐stimulating hormone secretion. Free inhibin/activin subunits and their dimers may also play an autocrine/paracrine role in the development of the testis and spermatogenesis. Furthermore, early onset of the α‐subunit in duck testes indicates that it may have an autocrine/paracrine effect on steroid hormones, which is important for sex differentiation.     

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.     

Comparative pharmacokinetics of florfenicol in healthy and Pasteurella multocida‐infected Gaoyou ducks

Pasteurella multocida is the causative agent of fowl cholera, and florfenicol (FF) has potent antibacterial activity against P. multocida and is widely used in the poultry industry. In this study, we established a P. multocida infection model in ducks and studied the pharmacokinetics of FF in serum and lung tissues after oral administration of 30 mg/kg bodyweight. The maximum concentrations reached (C_max) were lower in infected ducks (13.88 ± 2.70 μg/ml) vs. healthy control animals (17.86 ± 1.57 μg/ml). In contrast, the mean residence time (MRT: 2.35 ± 0.13 vs. 2.27 ± 0.18 hr) and elimination half‐life (T_½β: 1.63 ± 0.08 vs. 1.57 ± 0.12 hr) were similar for healthy and diseased animals, respectively. As a result, the area under the concentration curve for 0–12 hr (AUC_0–12 hr) for FF in healthy ducks was significantly greater than that in infected ducks (49.47 ± 5.31 vs. 34.52 ± 8.29 μg hr/ml). The pharmacokinetic differences of FF in lung tissues between the two groups correlated with the serum pharmacokinetic differences. The C_max and AUC_0–12 hr values of lung tissue in healthy ducks were higher than those in diseased ducks. The concentration of FF in lung tissues was approximately 1.2‐fold higher than that in serum both in infected and healthy ducks indicating that FF is effective in treating respiratory tract infections in ducks.     

Role of the ejaculatory bulb in biosynthesis of the male pheromone cis‐vaccenyl acetate in Drosophila melanogaster

In Drosophila melanogaster, the male ejaculatory bulb is the site of synthesis of a male‐specific pheromone, cis‐vaccenyl acetate, which functions as both an attractant and an anti‐aphrodisiac. This long monounsaturated acetate is structurally similar to a number of shorter gland‐synthesized moth pheromones. The cell monolayer that forms the Drosophila male ejaculatory bulb wall is responsible for the production and secretion of cis‐vaccenyl acetate into the seminal fluid. When dissected bulbs were incubated with sodium [14‐C]‐acetate (or deuterated acetate), a labeled acetate ester was synthesized. The labeled acetate ester co‐migrated with cis‐vaccenyl acetate in thin layer chromatography. Incubation of the abdomens of males from which the ejaculatory bulbs had been removed, or the abdomens of females, with radiolabeled acetate did not yield any acetate ester, but did yield other lipid products, including hydrocarbons. When the isolated labeled acetate ester was hydrolyzed, no radioactive vaccenol was formed. This strongly suggests that the acetyl group is incorporated via a transacetylation reaction, but that the vaccenyl moiety is not synthetized in the blub. The transacetylation enzyme activity was localized in the microsomal subfraction of the bulb homogenate, and its affinity for vaccenol was not very different from that reported for monounsaturated alcohol substrates in moths.