Immunoglobulin M associated with secretory component and immunoglobulin A deficiency in bovine colostrum.

Immunoglobulin (Ig) M purified from bovine colostrum was examined by an immunodiffusion analysis with antisecretory IgA serum and was found to be associated with a secretory component. Some of the combined proteins were dissociated if treated with 5 M guanidine-HCl and others were not. Another immunodiffusion analysis of 23 specimens of colostrum led to the finding that certain colostrums were deficient in IgA, even though they contained IgG and IgM.     

Quantitation and localization of immunoglobulin E containing cells in bovine lymphoid tissues

Although recent studies have begun to describe and quantify IgE responses in bovine serum and secretions, little is known about the distribution and quantity of IgE containing cells in cattle. In the present study, cells with cytoplasmic IgE were quantitated in bovine lymphoid tissues, using immunoperoxidase staining and evaluation by an image analysing computer (Quantimet). Frozen sections from retropharyngeal, bronchial and mesenteric lymph nodes, tonsil and spleen were stained from 11 calves, some of which had been exposed to antigen by aerosol or injection. Although individual variability was considerable, bronchial and mesenteric lymph nodes generally contained the greatest percentage of IgE containing cells, while retropharyngeal lymph node, tonsil, and spleen had less. Parenteral immunization with ovalbumin appeared to increase the splenic percentage, while aerosol exposure to ovalbumin was associated with a greater percentage of IgE containing cells in bronchial lymph nodes. Comparison of the present results with those reported for other species shows some similar trends in IgE localization.     

Use of a pilocarpine-based lavage procedure to study secretory immunoglobulin concentration in the alimentary tract of White Leghorn chickens.

A lavage procedure was used to study the kinetics of alimentary fluid IgA concentration in 15 specific-pathogen-free white leghorn chickens for 8 weeks post-hatch. Lavage solution was administered orally and collected from the distal alimentary tract following an intraperitoneal injection of pilocarpine. Concentrations of IgA, quantitated by enzyme-linked immunosorbent assay, were more than 0.04 mg/ml by 3 weeks and were negligible before this age. This level gradually increased over the next 5 weeks, peaking at nearly 0.4 mg/ml at 8 weeks of age. Alimentary lavage was easy to perform, required no necropsy or surgical manipulation, and facilitated repeated collection of alimentary fluid from live birds. Repeated lavage did not alter concentrations of IgA and IgG in alimentary fluid, and concentrations of IgA and IgG in alimentary fluid were stable during incubation at 37 C for 24-48 hr.     

The structure of bovine secretory component

Bovine secretory component (SC) has been cleaved with trypsin into a series of fragments and their N-terminal amino acid sequences have been determined. The close homology with the known sequence of human SC has enabled the sequential order of the fragments to be deduced. The results indicate that bovine SC consists of a single glycosylated polypeptide chain (Mr 74,000) folded into five globular immunoglobulin-like domains. A protein (Mr 94,000) has been isolated from detergent solubilised bovine epithelial membranes from liver, intestine and mammary gland. This membrane protein is specific for the binding of J-chain linked IgM and IgA dimers. It can be proteolytically cleaved into a water soluble SC-like portion and a detergent soluble hydrophobic portion. Bovine SC is therefore most likely to be the extracellular part of an epithelial receptor which mediates the transport of IgA dimers to mucosal surfaces. The various tryptic fragments from bovine SC have been shown to differ in their relative binding affinities for IgM and IgA dimers. The results imply that the first three domains of bovine SC are most involved in binding and domains 4 and 5 play subsidiary roles. Computerized prediction and modelling methods have been used to deduce possible tertiary and quaternary structures for SC. There are good indications that the molecule has an elonaged "zig-zag" structure stabilized by longitudinal inter-domain contacts. A model of SC bound to IgA dimer is presented.     

Commensal bacteria coated by secretory immunoglobulin A and immunoglobulin G in the gastrointestinal tract of pigs and calves

A large amount of secretory immunoglobulin A (S‐IgA) is secreted in the alimentary tract of mammals. It has been reported that S‐IgA coats a portion of commensal intestinal bacteria in human and mouse. However, S‐IgA‐coated bacteria have not been studied in pigs and calves. In this study, we evaluated the distribution of S‐IgA‐coated commensal intestinal bacteria in each portion of the gastrointestinal tracts of pigs and calves. Immunoglobulin G (IgG)‐coated bacteria were also analyzed because a considerable amount of IgG is secreted in the gastrointestinal tracts of pigs, and in particular, calves. S‐IgA‐ or IgG‐coated bacteria were detected in all the segments of the gastrointestinal tracts of pigs and calves. The proportion of S‐IgA‐coated bacteria to total bacteria (i.e. S‐IgA coating ratio) varied in the segments of the gastrointestinal tract in pigs, whereas those of calves were nearly the same throughout the gastrointestinal tract. The S‐IgA and IgG coating ratios were higher in pigs than in calves for all segments of the gastrointestinal tract.     

Distribution of hexoestrol residues in caponised chickens

A radioimmunoassay method is described for the detection and measurement of residues of hexoestrol and other stilbenes in tissues of poultry. The residues following caponisation with 12 mg hexoestrol were measured in leg muscle, liver, visceral fat and neck 44 days after implantation. They were significantly greater than values obtained from eight untreated control birds. The mean values obtained in the caponised chickens ranged from 471 pg/g net weight of leg muscle to 584,500 pg/g of tissue from the upper neck region, which included the site of implantation. Control values in untreated birds fell within the range of 8 pg/g in leg muscle to 44 pg/g in liver tissue.     

Germ cells and transgenesis in chickens

Chickens have proven to be useful organisms for transgenic research. This work provides enormous benefits in advancing animal biotechnology and aids in the development of unique technologies for bioreactor production and experimental model development. The various advantages of chicken transgenesis are derived from the genetic and physiological characteristics of this organism, although several physiological properties have impeded the development of an efficient transgenic system. We have developed embryo-mediated and testis-mediated transgenic systems using chicken primordial germ cells (PGCs) from embryos and testicular cells from adult males. These methods are efficient and involve minimal technical effort. Here, we review previous transgenic research using PGCs and testicular cells from chickens. Furthermore, we have summarized the development of the chicken model system in biomedical science and biotechnology and our recent achievements in this field.     

Influence of Campylobacter jejuni cecal colonization on immunoglobulin response in chickens

The immunoglobulin response of chickens to colonization by Campylobacter jejuni isolates B-540 and Clin-1 was monitored. Chicken humoral IgG and biliary secretory IgA (sIgA) responses were assessed by enzyme-linked immunosorbent assay (ELISA). Samples were taken from 128 C. jejuni-colonized chickens and 104 uncolonized chickens housed in a controlled environment. An indirect ELISA was performed using the homologous isolate of C. jejuni as the capture antigen and was developed with the specific goat anti-chicken IgG or IgA alkaline phosphatase conjugates. The ELISA absorbance values of the test samples at 405 nm (serum diluted 1:32 and bile diluted 1:10) were normalized in direct proportion to standard sera and bile sample values. In the colonized chickens, humoral IgG activities were highest at hatch, dropped to their lowest level after 2 weeks, and increased by 8 weeks to levels similar to those detected at hatch. The sIgA activity was lowest at hatch and increased by 4 weeks in colonized chickens while remaining lower in the control chickens. Chickens colonized with isolate B-540 showed a primary sIgA response during the first 4 weeks and reached a plateau over the final 4 weeks. In spite of these limited humoral and secretory immunoglobulin responses, once the chicken ceca was colonized by C. jejuni, the organism persisted throughout the 8-week experiment.     

Caecal microbiota of chickens fed diets containing propolis

The present study aimed to evaluate the effect of different levels of ethanolic extract of propolis (EEP) and raw propolis (RP) on broiler performance and on selected bacterial groups in caecal microbiota using fluorescent in situ hybridization (FISH) measured by fluorescent activated cell sorting. Two experiments were conducted with 120 male chicks from 1 to 21 days of age for each, raised in cages and distributed in a completely randomized experimental design; there were five replicates with four birds per experimental unit and six treatments for each experiment (trial 1 – EEP – 0, 1000, 2000, 3000, 4000 and 5000 ppm and trial 2 – RP – 0, 100, 200, 300, 400 and 500 ppm). Fluorescent probes were used against the bacterial groups in caecal samples collected at 21 days of age. The data were subjected to one‐way anova followed by Tukey's and regression analyses were used to assess the relationship between dietary levels of EEP or RP on performance and intestinal microbiota (p < 0.05). In the trial 1, results showed that the EEP did not cause any significant (p > 0.05) modification in the performance and caecal microbiota. In the trial 2, RP inclusion did not affect the performance but changed the bacterial composition (p < 0.05). Clostridiaceae, Gammaproteobacteria excluding Enterobacteriaceae and Lactobacillus spp. showed a quadratic response (p < 0.05), with the lowest value predicted to occur at 240 ppm, 221 ppm and 213 ppm of RP respectively. The proportion of Bacteroidaceae and Gammaproteobacteria did not differ (p > 0.05) among the experimental groups. The inclusion of ethanolic extract of propolis did not affect the performance and intestinal microbiota, whereas the supplementation of raw propolis modulates the caecal microbiota composition without any effects on chicken performance.     

Feeding behaviour in chickens given diets containing medium chain triglyceride

1. The effect of dietary medium chain triglyceride (MCT) on short‐term food intake was compared with the effect of long chain triglyceride (LCT) in chickens. Maize oil was used as the LCT while glyceryl tricaprylate (C 8) and glyceryl tricaprate (C 10) were used as MCT. Cumulative food intake was determined during the 6 h after the start of feeding.

2. Chicks were given diets containing 200 g C 8/kg diet, 200 g C 10/kg diet or 200 g LCT/kg diet in experiment 1. As early as 30 min after feeding, cumulative food intake in both MCT‐supplemented diets decreased significantly compared with the diet containing LCT.

3. To determine if endogenous cholecystokinin (CCK) was responsible for the decrease in food intake caused by MCT, birds were injected with the CCK‐A receptor antagonist devazepide (DVZ, 1 mg/kg BW) before diet presentation. DVZ had no effect on food intake with either LCT‐ or MCT‐supplemented diets.

4. In experiment 3, chicks were given a choice between either diets containing LCT and C 8, LCT and C 10, or C 8 and C 10 to confirm whether or not the palatability of the diets was influenced by the dietary fat sources. There was no difference in food intake between C 8 and C 10‐supplemented diets. However, chicks preferred the LCT‐supplemented diet compared with either of the diets containing MCT.     

Bile and serum immunoglobulin levels during primary and secondary infections with Eimeria tenella in chickens

Using a radial immunodiffusion assay, total bile and serum IgG, IgM and IgA were measured following primary and secondary exposures to Eimeria tenella. Neither IgG nor IgM could be detected consistently in bile. Biliary IgA peaked at Days 6 and 10 following a primary infection of either 5000 or 10,000 oocysts and remained elevated following a subsequent 10,000-oocyst challenge at Day 10. Serum IgG and IgM levels were not influenced by parasitism and measurable concentrations of serum IgA were not detected.     

Distribution of protoporphyrin in female broiler chickens affected with protoporphyria

One hundred thirty-seven broiler chickens at a poultry meat processing plant had dark green to black livers. Thirty-one chickens of these were collected at random and examined pathologically and biochemically. All of thirty-one chickens were female. The chickens showed mild retarded growth and a remarkable atrophy of the gallbladders. Microscopically, the livers showed dark brown pigments in the Kupffer cells, hepatocytes, and portal triads. These pigments showed birefringence with a Maltese-cross pattern under polarized light. Hyperplasia of the cholangioles, fibrosis, and infiltration of inflammatory cells were present in the portal triads. All the examined samples showed the same dark brown pigments in alveolar walls of the lungs. A high concentration of protoporphyrin was detected in affected livers, marrow, and feces (489, 104, and 116 microg/g wet wt., respectively) by biochemical assay.     

鸡睾丸细胞分离方法与培养的研究

近年来,随着对睾丸细胞研究的进一步深入,睾丸细胞的起源及其生成精子的机制已经明了,并且对它们的分离和体外培养也已取得了阶段性的进展。但这些研究多限于一些哺乳类动物,尤其以啮齿类动物居多[1]。而在禽类方面的研究还处于启蒙阶段。本研究旨在通过对鸡睾丸细胞几种分离方