Architecture of sheep haemal nodes

Sheep haemal nodes are reported to be lymphatic organs within the blood circulation. In this study they were seen to be covered by a capsule of connective tissue, with a hilus in which some blood vessels were detectable. Below the capsule a subcapsular blood sinus and lymphatic nodules were present. A reticular meshwork extended through the interior; the interstices of the reticular meshwork contained a large number of free blood cells, many macrophages, lymphocytes and plasma cells. Blood collected in the subcapsular sinus and flowed through the reticular meshwork aided by the particular alignment of some of the reticular cells in channel-like passageways. Venules originated in the parenchyma and drained into efferent veins that left the organ at various points in the capsule. A large efferent blood vessel, apparently connected with the subcapsular sinus, was present in the hilus.     

A Scanning and Immunohistochemical Study in Bovine Haemal Node

The bovine haemal nodes are lymphatic organs with haemal circulation. The blood circulates inside of them through virtual cavities named sinuses. In these sinuses there is passage of cells and particulate materials from parenchyma to sinuses and vice versa. For this passage temporal overtures exist in the sinus walls. The sinus wall is composed of endothelial cells, basal membrane and reticular cells. Our results have demostrated that type IV collagen was one of the basal membrane constituents in bovine haemal nodes and by this constitution a dynamic equilibrium of the sinus walls was permitted.     

Morphologic features of lymph vessels in caprine hemal nodes

The distribution and characteristics of lymph vessels in caprine hemal nodes were studied after glutaraldehyde fixation and epoxy resin embedding. Histologically, the lymph vessels were characterized by thin walls and wide lumens containing inspissated lymph in which a few cells were suspended. The lymph vessels contrasted sharply with adjacent blood sinuses that were filled with elements of circulating blood. A circumferential lymph vessel in the cortex joined radial branches in the medulla that met at the hilum to drain through a large efferent lymph channel. Ultrastructurally, the lymph vessel wall comprised endothelial cells supported by a continuous basal lamina, collagen fibrils, and adventitial reticular cells. The cytoplasm of endothelial cells had fenestrations, plasmalemma-associated vesicles, vacuoles, and focal splits that enclosed large compartments. Many compartments contained erythrocytes and lymphocytes. Modifications of the endothelial cells signified their endowment with features that favored transendothelial transport. The distribution of lymph vessels and the finding of only efferent lymph channels are related to the roles of hemal nodes in blood storage by hemoconcentration and in immune defense mechanisms.     

Lesions in clinically healthy cattle persistently infected with the virus of bovine viral diarrhea--glomerulonephritis and encephalitis

Four clinically healthy cattle persistently infected with the virus of bovine viral diarrhea were examined for viral antigen and lesions. Antigen was seen by direct immunofluorescence in cytoplasm of the neurons of the brain and cervical part of the spinal cord, cells and basement membrane of renal glomeruli, reticular cells of lymph nodes and spleen, epithelial cells of small intestinal crypts and renal and testicular tubules, and endothelial cells of blood vessels. Infected neurons were pyknotic and surrounded by astrocytes and macrophages. A few blood vessels in the brains were cuffed with mononuclear cells. Basement membranes of renal glomeruli were irregularly thick with eosinophilic material, and mesangial cells in the glomeruli were plentiful. The virus had a direct effect on some tissues, but was restricted in its cytopathogenicity and was not eliminated by defense mechanisms of the host. Renal glomerular lesions were believed to have an immunologic basis.     

Histological, histochemical and immunohistochemical study of the haemal nodes of the dromedary camel

Haemal nodes are lymphoid organs found in various mammals and some birds. The structure of haemal nodes has been described in a number of species but not yet in the camel. Therefore, haemal nodes from 10 camels were studied histologically and tested for CD3, CD22, major histocompatibility complex (MHC) class II/DR, alpha-smooth muscle actin and for the demonstration of acid and alkaline phosphatases. The haemal nodes were of spherical or kidney shape with one or two hili and had a capsule and trabeculae of connective tissue and smooth muscles. The main parenchyma was composed of a cortex and a medulla. The cortex was formed from lymphoid follicles and diffuse interfollicular lymphocytes. The medulla consisted of lymphoid cords separated by medullary sinuses. The interfollicular lymphocytes and those in the medullary cord were CD3-positive. The lymphoid follicles showed CD22-positive cells. MHC class II/DR was expressed by most cells of the parenchyma. There were also subcapsular, peritrabecular and medullary blood sinuses. Afferent and efferent lymphatics and lymphatic sinuses were also found. Acid phosphatase-positive cells were localized mainly in the marginal, the interfollicular zone and in the medullary cord. Alkaline phosphatase positivity was observed in the endothelium of the sinuses and in the lymphoid follicles. The morphology of these organs in the camel allows a classification as haemolymph nodes and suggests involvement in blood and lymph filtration.     

IL1RN基因在晋汾白猪仔猪和新山西黑猪仔猪不同组织和细胞中的表达特征分析

白细胞介素1受体拮抗剂（interleukin 1 receptor antagonist,IL1RN）是一种重要的宿主免疫调节因子,是天然存在的IL-1拮抗剂。为了解IL1RN基因在断奶仔猪不同组织和细胞中表达的特征,利用Trizol法提取晋汾白猪仔猪和新山西黑猪仔猪的心脏、肝脏、脾脏、淋巴细胞、巨噬细胞等18种组织和细胞的总RNA,采用RT-PCR方法检测IL1RN基因在不同组织和细胞中的表达情况,应用SPSS 19.0软件分析IL1RN基因在晋汾白猪仔猪和新山西黑猪仔猪4种免疫组织和细胞（脾脏、淋巴结、巨噬细胞、白细胞）中的表达差异情况。结果显示：IL1RN基因在晋汾白猪仔猪和新山西黑猪仔猪的脂肪组织中均不表达,在其余各组织和细胞中均表达;IL1RN基因在晋汾白猪仔猪的巨噬细胞与淋巴结、脾脏、白细胞之间的表达差异极显著（P<0.01）,且在淋巴结与脾脏、白细胞之间的表达差异极显著（P<0.01）;IL1RN基因在新山西黑猪仔猪的脾脏、淋巴结、巨噬细胞、白细胞的表达两两之间均呈极显著差异（P<0.01）;IL1RN基因在2个猪种的脾脏、巨噬细胞、白细胞之间的表达呈极显著差异（P<0.01）,在淋巴结中的表达呈显著差异（P<0.05）。该研究结果为进一步揭示2个猪种的IL1RN基因在疾病抵抗方面的作用及相关机制提供了依据。     

Effects of the adjuvant DEAE-dextran and Staphylococcus aureus on the lymph pathways and filtering capacity of popliteal lymph nodes in sheep

The effects of subcutaneous injections of the adjuvant DEAE-dextran and, or killed Staphylococcus aureus on the structure of the lymph pathways of popliteal lymph nodes in sheep were examined using light and electron microscopy and Microfil casts. Dextran with or without killed S aureus caused significant changes in the lymph pathways both within the node and outside it. However, killed S aureus alone did not. The changes included anastomoses among afferent lymph vessels and between afferent and efferent lymph vessels; proliferation of vessels around the node; joining of parts of the capsule and trabeculae to adjacent parenchyma with loss of parts of the subcapsular and trabecular sinuses; enlargement of medullary sinuses; and reduction of the number of reticular processes in sinuses throughout the node. These changes were accompanied by a reduced ability of the node to filter chicken red blood cells labelled with chromium-51 which were injected into an afferent lymph vessel.     

Tacrolimus-FK506 induced immunosuppression in gnotobiotic piglets

Tacrolimus (FK506), an inhibitor of calcineurin, is an immunosuppressive agent used in clinical trials of transplant patients. Although FK506 targets Ca(2+)-mediated T-cell signaling, phenotype(s) of the specific target cells and the corresponding cytokine pathways are not well known. In this study, the impact of FK506 on number and characteristic of T-cells in selected lymphoid tissues of gnotobiotic (GB) piglets was determined. FK506-treated GB piglets were compared with untreated GB and conventional piglets. The T-helper, cytotoxic, natural killer, double-positive, and activated T-cell populations were analyzed in suspensions of mononuclear cells isolated from thymus, mesenteric lymph nodes and peripheral blood. In vitro secretion of interleukin-8 and interferon-gamma in concanavalin A-stimulated lymphoid cell-cultures was measured by ELISA. Daily intramuscular treatment of GB piglets with 1mg/kg of FK506 from birth for 4 weeks resulted in lowered (P<0.05) in vitro secretion of interferon-gamma and interleukin-8. Moreover, depletions of MNC in systemic and mucosa-associated lymphoid tissues were observed in piglets treated with FK506. The depletions of mononuclear cells and low levels of interferon-gamma and interleukin-8 in piglets treated with FK506 were accompanied by lower proportion of CD3+, CD2+CD4+ and CD2+CD8+ T-cell phenotypes in peripheral blood but not in thymus and mesenteric lymph nodes. These results indicate that FK506-treatment causes immunosuppression in GB piglet, and this effect could be exploited further to study opportunistic pathogens in pig model.     

Transfer of humoral and cell-mediated immunity via colostrum in pigs

The first aim of our study was to obtain information on the transmission of antigen-specific antibodies from colostrum to respiratory tract mucosa in piglets. The second aim was to confirm the biological relevance of the presence of lymphocytes in colostrum and the already described fact that these cells can penetrate the intestinal barrier and "colonize" peripheral blood and lymphatic tissues of piglets. Therefore, we performed an experiment in which sows were immunized with a model antigen Keyhole Limpet Hemocyanin and their piglets were euthanized at different intervals after birth and colostrum intake. Colostrum, bronchoalveolar lavage fluid and blood samples were collected for serological detection of antigen-specific antibodies. Lymphocytes isolated from peripheral blood and lymphatic tissues (mesenteric and tracheobronchial lymph nodes and spleen) of piglets were in vitro activated with the antigen. We found that colostrum-derived antibodies can cross into the respiratory tract mucosa. Furthermore, we found that antigen-specific lymphocytes were detectable in mesenteric lymph nodes and peripheral blood, but very rarely in spleen and tracheobronchial lymph nodes.     

The effect of intramuscular administration of colistin on the development and course of experimentally induced oedema disease in weaned piglets

Shiga-toxigenic E. coli (STEC) strains that produce Shiga toxin Stx2e cause oedema disease in weaned piglets. The purpose of the present study was to investigate the impact of Stx2e released in mesenteric lymph nodes on disease pathogenesis. Colistin and ampicillin were intramuscularly administered to piglets of the experimental group simultaneously challenged with STEC strain, type O139:F18ab, Stx2e+. Piglets of the control group were challenged with STEC only. The strain was naturally resistant to ampicillin and susceptible to colistin. After the challenge, colonisation of the intestines was observed in both antibiotic-treated piglets and control piglets without antibiotic treatment. Histochemistry and scanning electron microscopy revealed sporadic colonisation of the small intestine in the piglets. STEC was detected in the mesenteric lymph nodes of untreated piglets. The clinical manifestations of oedema disease were observed in both groups. In the antibiotic-treated group (11 piglets), oedema disease developed in 10 piglets, eight of which died or were euthanized ante finem. In the untreated group (11 piglets), oedema disease developed in five piglets, four of which died or were euthanized ante finem. We therefore propose that the STEC lysed by colistin suddenly released the toxin from bacterial cells immediately after their passage through the intestinal wall. That could explain a more severe course of oedema disease in the treated piglets. Even though high amounts of STEC were present in the lymph nodes of untreated piglets, the toxin was not released abruptly because the bacterial cells were not damaged.     

STUDIES ON THE PATHOGENESIS OF BOVINE EPHEMERAL FEVER

A study of the pathogenesis of bovine ephemeral fever confirmed that the major clinical signs were fever lasting no more than 2 days, with increased respiratory rate, dyspnoea and some degree of lameness. Haematological observations revealed a neutrophilia with a left shift and a lymphopaenia at the time of peak clinical reaction. The net result was a slight leucopaenia on the day after this reaction. The most prominent pathological changes involved the lungs and synovial joints. Pulmonary emphysema and alveolar collapse with bronchiolitis, degenerative changes in synovial membranes and increased synovial fluid were observed. Specific fluorescence indicating the presence of BEF viral antigen could be detected at the time of peak clinical response in individual cells in the lungs, spleen and lymph nodes as well as neutrophils. Before and after the peak fever some fluorescence was seen in cells which appeared to be reticular cells in the lymph nodes. Viral isolation in mice could be made from blood, lungs, spleen and lymph nodes over a period of no more than 3 days. It is postulated that viral growth takes place mainly in the reticuloendothelial cells in the lungs, spleen and lymph nodes and not in vascular endothelium or lymphoid cells.     

Blood vessels within lymph nodes: a comparison between pigs and sheep

In sheep, most blood vessels enter the lymph nodes at a discrete hilus, which is where the capsule overlies medullary tissue. In pigs, vessels within the node arise from an extensive network of arteries on the surface, and most vessels penetrate the capsule where it overlies diffuse tissue--the counterpart of the sheep medulla. In both species the blood vessels divide extensively as they approach and supply the dense lymphoid tissue which contains lymph nodules. The blood vessels within sheep lymph nodules are surrounded by a thick layer of amorphous material. The dense lymphoid tissue has an abundant network of venules which have smooth muscle cells in their walls. These venules are involved in lymphocyte migration and in pigs they are lined by high endothelium which is similar to that in most other species, while in sheep the endothelium is much lower. Fenestrated capillaries occur in both pig and sheep lymph nodes.     

Pathogenicity of Isospora suis in gnotobiotic and conventionalised piglets

Isospora suis is unequivocally a primary pathogen of swine. Inoculation of I suis in conventionalised and germ-free piglets caused a biphasic disease course with marked diarrhoea, villous atrophy and necrosis of the intestinal epithelium at four to six and eight to 10 days after inoculation. The presence of a normal bacterial flora markedly (P less than 0.05) influenced the survival rate of piglets but did not appear to affect the histopathological changes observed. Mild limited focal necrosis and bile stasis were present in the liver at eight to 10 days after inoculation. In this period there was also ectasia of lymph vessels in the intestinal lymph nodes.     

Lymphocyte proliferation in lymphoid organs of the dromedary camel using the monoclonal antibody MIB-5 against the proliferation-associated nuclear epitope Ki-67

Detection of proliferating lymphocytes is useful for studying immune reactions and for the prognosis of tumours of lymphocyte origin. Markers detecting proliferating cells are lacking in the dromedary camel. This study deals with the immunohistochemical detection of the Ki-67 proliferation-associated nuclear epitope using MIB-5 in frozen sections from spleens, different lymph nodes and haemal nodes of eight camels (0.5-12 years old). Frozen sections from rat spleens were labelled in parallel with camel tissue as a positive control. Large numbers of cells expressing the Ki-67 epitope were localized in germinal centres of all lymphoid organs tested. A few cells were found in the periarterial lymphoid sheath and the red pulp of the spleen, also in the lymphatic cords of the lymph nodes and the haemal nodes. There were no obvious differences in respect to the age of the animals. The Ki-67 epitope is well expressed by proliferating cells in camel lymphoid organs. MIB-5 can be applied to identify this epitope and will be a useful marker for cell production in immune reactions.     

The Thymus of the Guinea Fowl from the Eighteenth Day of Incubation until Maturity

The thymus gland of the guinea fowl appeared as a series of pink, irregular-shaped lobes along the jugular veins. These lobes were 13 in number with seven on the right and six on the left side. The third and sixth lobes on the right side were the largest. The largest lobe measured about 20.0 × 5.0 mm while the smallest measured about 5.0 × 4,0 mm. Histologically, it was encircled by a thin layer of poorly stained connective tissue from which strands penetrated the thymic tissue giving incomplete lobules peripherally as early as day 18 of incubation. Lymphocytes and reticular epithelial cells were the two types of cells occurring at this time. Reticular epithelial cells were observed to be vacuolated especially towards the centre of the gland. Macrophages were observed in the thymus on day 20 of incubation for the first time. The blood vessels had accompanied the interlobular septa into the thymus. By day 21 of incubation, blood vessels had actually penetrated the thymic tissue but most of these vessels were of the capillary size. The cortex also became quite distinct from the medulla. At day 22, vacuolation ceased to appear in the cells at the cortical zone. Macrophages were still less than 10 in the slide. At day 23, most blood vessels were within the medulla and they displayed compressed lumi-na. Mitotic figures became a common site amongst the large lymphocytes and reticular epithelial cells. By day one post-hatch, lobulation of the thymus was still poor and macrophages became difficult to identify. The small and medium lymphocytes had outnumbered the large lymphocytes throughout the thymic tissue except at the most peripheral zone of the cortex. Fine vacuoles had reappeared within the cytoplasm of the reticular epithelial cells in the cortex. By day five, the distribution of blood vessels was almost uniform between the cortex and medulla. At three weeks post-hatch, plasma cells and red blood cells were seen in the stroma of the gland for the first time and most of the red blood cells occurred in the medulla.     

Histology and Ultrastructure of the Lymph Nodes of the Buffalo (Bos bubalus)

Pre‐scapular, femoral and mesenteric lymph nodes from five buffalo calves and five buffalo bulls were studied using light and transmission electron microscopy. The nodes were surrounded with a thin capsule of dense connective tissue and smooth muscles. Subcapsular and trabecular lymphatic sinuses were lined with endothelial cells resting on a basement membrane. The cortex was formed by lymphoid follicles and inter‐follicular lymphocytes. Primary and secondary follicles were observed. The medulla was made up of medullary cords of lymphocytes separated by lymphatic sinuses. These sinuses were lined with a discontinuous epithelium and interestingly crossed by reticular fibres. High endothelial venules were found in the paracortical area. Several lymphocytes were observed infiltrating the wall of these venules. The lymph nodes of the Egyptian water buffalo showed a typical structure compared with the majority of mammals, with no age‐related structural variation.     

实验性流行性腹泻仔猪小肠粘膜上皮细胞及肠系膜淋巴结的超微结构

给8头生后3d的哺乳仔猪经口感染猪流行性腹泻病毒(PEDV)“吉”毒株,于感染后18、30、45和96h各扑杀2头,以透射电镜和扫描电镜观察了小肠粘膜上皮细胞及肠系膜淋巴结的超微结构。结果表明,小肠上皮细胞的病变因感染时间不同而有明显差异。上皮细胞的脱落和残留上皮细胞超微结构的破坏,以感染后30h最严重,病毒在这些上皮细胞内的增殖最显著。感染后45h,见有大量新生上皮细胞修补损伤的肠绒毛。感染后96h,小肠绒毛短缩、粗大乃至发生融合。实验仔猪肠系膜淋巴结内巨噬细胞和淋巴细胞的超微结构均遭到破坏,在巨噬细胞内见有PED冠状病毒粒子。     

Longitudinal study of Salmonella infection in Italian farrow-to-finish swine herds

A longitudinal study of Salmonella enterica infection was carried out in five Italian farrow-to-finish swine herds previously known to be infected by Salmonella. Five litters were randomly selected from each herd and in each litter six piglets were randomly selected and individually identified. Thus, the study included 30 pigs from each farm. At weaning, individual blood samples were collected for serological examination from all selected piglets and on the same day from all sows in the farrowing unit. Piglets were bled again at approximately 60, 90, 150, 210 and 270 days of life whereas the last blood sample was collected at slaughtering. In one of the herds, in which the duration of productive cycle was about 12 months, the last blood samples were collected at 350 days of life. With the same time scheduling, five pen pooled faecal samples were collected from each herd for bacteriological examination. At slaughtering, mesenteric lymph nodes were collected from each ear-tagged pig. Sero-prevalence (cut off S/P ratio 0.25) in sows varied from 93.8% to 100%. In four herds, sero-prevalence in piglets showed a similar profile with complete decline of maternal antibodies at day 60 and clear sero-conversion between day 90 and day 150. In one herd, sero-conversion was observed earlier and 56% of piglets were positive at day 90. The peak of sero-prevalence was observed between day 210 and day 270. Sero-prevalence at slaughtering varied from 66% to 100%. Salmonella was isolated from faecal samples in four of five herds. No Salmonella was isolated from mesenteric lymph nodes at slaughter in two of the herds. Culture prevalence from mesenteric lymph nodes in the other three herds ranged from 3.3% to 30%. This longitudinal study provides original information about epidemiological dynamics of Salmonella enterica infection in Italian swine herds in consideration of the unique extended fattening period typical of the Italian production.     

不同毒力猪繁殖与呼吸综合征病毒对仔猪肺和外周免疫器官损伤的研究

为研究不同毒力的PRRSV对仔猪肺脏和外周免疫器官损伤的差异,本实验分别采用PRRSV变异株(HuN4株)和PRRSV经典株(CH-1a株)感染35日龄健康的断奶仔猪,并在感染后0 d、3 d、7 d、10 d和14 d各迫杀3头,检测肺、颌下淋巴结、肠系淋巴结、腹股沟淋巴结、扁桃体和脾脏的病毒载量及病理变化情况,同时检测血清中抗PRRSV的抗体水平。结果表明:感染后3 d肺脏及各免疫器官可检测到病毒,HuN4感染组病毒载量比CH-1a感染组病毒载量高1 000倍;HuN4感染组病毒载量峰值出现在感染后10 d,而CH-1a感染组维持着较低水平的病毒载量。组织病理学检测显示HuN4感染组淋巴结内淋巴细胞显著减少,呈空泡状;CH-1a感染组淋巴结内淋巴细胞轻度减少,呈星隙状。本实验表明HuN4株比CH-1a株对肺和外周免疫器官造成更严重的损伤。     

Activation of the immune system is the pivotal event in the production of wasting disease in pigs infected with porcine circovirus-2 (PCV-2)

Porcine circovirus (PCV)-2, a newly described single-stranded circular DNA virus pathogen of swine is the cause of postweaning multisystemic wasting syndrome (PMWS). In gnotobiotic piglets, PCV-2 infection alone produces asymptomatic infection without evidence of overt PMWS. Gnotobiotic piglets infected with PCV-2 were injected with keyhole limpet hemocyanin in incomplete Freund's adjuvant (KLH/ICFA), and the effects on virus production and development of PMWS were determined. In the first experiment, piglets were injected subcutaneously on the left hip and shoulder, and viral burden was assessed in regional lymph nodes draining the injection sites and in contralateral lymph nodes 13-14 days after infection. Immune activation increased the number of virus antigen-positive cells in draining lymph nodes and increased the amount of infectious virus recovered by 1-4 log10. In a second experiment, the effects of injections of KLH/ICFA with or without concurrent stimulation of peritoneal macrophages by intraperitoneal injections of thioglycollate broth on induction of PMWS was assessed. All immunized piglets developed moderate to severe PMWS, whereas none of the piglets infected with PCV-2 alone developed PMWS. In PMWS-affected piglets, extensive replication of PCV-2 was documented by both immunocytochemistry and quantitative viral titrations. Thus, immune activation is a key component of the pathogenesis of PCV-2-associated PMWS in swine.