Immunohistochemical Localization of Galanin in Bovine Reproductive Organs

The vagina, uterus and oviduct were shown to receive galanin immunoreactive (GAL-IR) nerve fibres, the number of which varied between particular organs. In the ovary, GAL-IR nerves were absent. A small number of these nerves were located in the layers of the oviduct. A moderate number of GAL-IR nerves were situated in the body and uterine horns, whereas the uterine cervix and vagina wall contained a large number of GAL-IR nerve fibres, evenly distributed throughout particular membranes of the organs. GAL-IR nerves were found to contain, simultaneously, either vasoactive intestinal polypeptide (VIP), substance P (SP) or Leu⁵-enkephalin (ENK). Many of the GAL-IR nerves contained tyrosine hydroxylase (TH). A group of GAL-IR nerves that did not possess immunoreactivity to VIP, SP, ENK or TH was also observed.     

An Immunohistochemical Survey of Catecholamine-synthesizing Enzyme-immunoreactive Nerves and Endocrine Cells in the Bovine Pancreas

The distribution of catecholamine-synthesizing enzyme-immunoreactive nerves and endocrine cells in the pancreas of the calf and cow was studied immunohistochemically using antisera against tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). TH- and DBH-immunoreactive nerve fibres were abundant both within and around the islet of Langerhans. A few TH- and DBH-immunoreactive nerve fibres were seen around the large islets characteristic of calf pancreas, but the majority of cells in the large islets, and some in islets of Langerhans, showed TH immunoreactivity. In the exocrine pancreas, both TH- and DBH-immunoreactive nerve fibres were distributed randomly among the acini, with the DBH-immunoreactive fibres being more numerous. Abundant TH- and DBH-immunoreactive nerve fibres were seen in close association with blood vessels and in the connective tissue around the interlobular duct. Immunoreactivity for both enzymes was also observed in the nerve cell bodies and fibres of the intrapancreatic ganglia. The findings suggest an important role for catecholamines in the regulation of bovine pancreatic function.     

Fractures of Ossified Rib Cartilages in the Horse – a Concern of Veterinarians?

The morphology, neurochemistry and function of intramural nerve structures in the mammalian gastrointestinal tract are relatively well known, but in normal, healthy individuals. The present study was aimed at investigating the chemical coding of nerve structures in the wall of the ileum and large intestine in normal pigs ( n  = 3) and in pigs undergoing dysentery ( n  = 6). Dysentery was evoked by artificial infection of the clinically healthy animals per os with Brachyspira hyodysenteriae . All the animals were deeply anaesthetized and transcardially perfused with 4% paraformaldehyde. The cryostat sections of the intestines were processed for double-labelling immunohistochemistry using antisera against PGP 9.5, GAL and VIP. In the intramural plexuses of the control pigs, the percentage of GAL-immunoreactive (GAL-IR) perykarya varied from 11% (descending colon) to 19% (centrifugal turns of the ascending colon) whereas in the dysenteric pigs, it was distinctly higher, reaching from 28% (ileum) up to 48% (cecum). In the control animals, the percentage of VIP-IR neuronal somata varied from 3% (descending colon) to 19% (ileum). In dysenteric pigs, it was from 6% (descending colon) up to 28% (cecum). In the muscular coat (MC) and mucous membrane (MM) of the normal intestine, very numerous GAL- and VIP-IR nerve fibres were observed. The nerve fibres in the myenteric plexus (MP) were even more numerous than those in the muscular coat while in the outer (OSP) and inner (ISP) submucous plexuses, they were less abundant. In the dysenteric pigs, the nerve fibres found in MC, MP and OSP were less numerous, whereas those observed in ISP and MM were more abundant than those in the control animals. The present results suggest that GAL and VIP are involved in the regulation of inflammatory processes developing in the porcine gastrointestinal tract during dysentery.     

Effects of testosterone on rat placental development

We investigated the morphological effects of testosterone on placental development in a rat model of polycystic ovarian syndrome (PCOS). Testosterone propionate (TP), which was subcutaneously administered to pregnant rats with 5 mg/animal from gestation day (GD) 14 to GD 18, induced a maternal weight reduction without mortality or clinical signs from GD 19 onwards. A decrease in fetal and placental weight, an increase in intrauterine growth retardation (IUGR) rates, and histological changes in the placenta were observed on GD 21 but not on GD15 or 17. Histopathologically, on GD 21, the trophoblast septa thickened, and the maternal sinusoids were narrowed in the labyrinth zone, resulting in a small placenta. Additionally, the placental weight, thickness, and histological morphology in the labyrinth zone on GD 21 in the TP-treated group were nearly identical to those on GD 17 in the control and TP-treated groups. Therefore, it was assumed that the testosterone-induced small placenta was induced in association with the developmental inhibition of the fetal part of the placentas from GD 17 onwards.     

Innervation of the circumvallate papilla of the cat tongue

Immunohistochemically, the distribution of S-100 protein and acetylated tubulin-positive nerve fibres was studied in the circumvallate papilla and its taste buds (TB) in the adult cat. The immunostaining for acetylated tubulin demonstrated an extensive innervation of the circumvallate papilla. Vegetative ganglionic cells were found in the central area of papilla, whereas fine nerve fibres were concentrated under epithelium. Individual positive axons were seen in relation to TB. Nerve fibres enter the TB and branch out among the supporting and sensory cells. Some nerve fibres reach the apical surface of the TB, including the taste pore. Nerve fibres positive for S-100 protein were observed as dense nerve plexus located in the core of the papilla. Satellite cells localized inside the papilla were seen to surround the ganglionic cells. Bands of fine nerve fibres were present under lining epithelium, mainly at the base of the TB. A weak reaction was displayed by taste bud cells and surrounding epithelial cells as well as by the epithelial cells of the papillary side of the moat. A dense network of the nerve fibres was present among the glandular acini and surrounding the ducts of the serous Ebner's glands.     

Lesions in the thymus and bone marrow in chicks with experimentally induced chicken infectious anemia disease

One-day-old SPF chicks were inoculated with the Cux-l strain of chicken infectious anemia virus (CIAV), and the clinical development of disease and its macroscopic and microscopic alterations in the thymus and bone marrow, were observed. Tissue sections of thymus and bone marrow were stained using the streptavidin-biotin peroxidase method and examined under light microscope for evaluation of antigenic intensities in tissues. Those findings were then compared with blood parameters and ELISA results obtained through collected sera during sacrifice procedures. We sought to determine: the localization of viral antigens in thymus and bone marrow tissues after inoculation, the correlation between antigen intensities and hematologic, serologic and histopathologic findings, definitive diagnostic criteria using histopathologic and immunoperoxidase methods, and the reliability of these methods in the diagnosis of CIAV infection. For this purpose, 83, one-day-old SPF chicks were used. The birds were divided into experimental (n = 52) and control (n = 26) groups. A virus dose of TCID50 of 100,000/ml was administered intramuscularly to every bird in the experimental group. Based on the results of this study, we have suggested that clinical examination, along with macroscopic and microscopic evaluation of the thymus and bone marrow, maybe undertaken starting from day 7 post-inoculation (PI). ELISA, might be of value, as it might give consistent results starting from day 14 PI. However, the most reliable results were obtained through examination of thymus and bone marrow sections from infected birds stained by immunoperoxidase technique, as early as day 4 PI.     

Phagocytic and nitroblue tetrazolium reductive properties of mature and immature neutrophils and eosinophils from blood and bone marrow from cows

Functional capabilities of morphologically mature (segmented) and immature granulocytes (neutrophils and eosinophils) from bone marrow from cows were studied and compared with similar activities of segmented granulocytes from blood. Phagocytosis of Escherichia coli and postphagocytic oxidative metabolic stimulation, measured by nitroblue tetrazolium (NBT) reduction, were evaluated simultaneously. Phagocytosis was observed readily in segmented neutrophils, neutrophilic bands, and metamyelocytes and rarely in myelocytes. Phagocytosis was not seen in promyelocytes and myeloblasts. Neutrophilic bands and metamyelocytes were phagocytically less active than were segmented neutrophils. Washed segmented bone marrow neutrophils possessed phagocytic activity similar to that of blood neutrophils, whereas the activity of unwashed segmented bone marrow neutrophils was markedly less than that of blood neutrophils. Reduction of NBT was observed only in blood segmented neutrophils and bone marrow segmented neutrophils; the magnitude of NBT reduction was significantly (P = less than 0.005) less in bone marrow neutrophils than in blood neutrophils. Eosinophils were phagocytically less competent than were neutrophils. The NBT reduction was observed only in eosinophils from blood, but not in eosinophils from bone marrow.     

鸡传染性贫血病毒感染胸腺集落刺激因子对骨髓粒单系祖细胞增殖功能的影响

本文研究了鸡传染性贫血病毒感染鸡胸腺ＣＳＦ对正常鸡骨髓粒单系祖细胞（ＣＦＵ—ＧＭ）增殖功能的影响．结果表明，雏鸡感染ＣＩＡＶ后１４天胸腺ＣＳＦ对ＣＦＵ—ＧＭ刺激作用与对照鸡比较未见明显异常．感染后２１天，刺激作用较对照鸡明显增强（ｐ＜０．０１）；感染２８天以后，其刺激作用达正常成水平．结果证明，ＣＩＡＶ感染引起的骨髓造血机能障碍与胸腺ＣＳＦ活性无关．     

Lesions of bone and bone marrow in myeloid leukosis occurring naturally in adult broiler breeders

Lesions of bone and bone marrow in myeloid leukosis (ML) occurring naturally in adult broiler breeders were investigated pathologically. During gross examination, nodules and protrusions were commonly observed on the surface of the sternum, ribs, vertebrae, and synsacrum. The bone marrow of all the bones of the body was pale in color. Histologically, granulated myelocytes proliferated in the bone marrow of various bones and in the periosteum of the sternum, ribs, vertebrae, and synsacrum. The first proliferation of tumor cells occurred in the bone marrow of epiphysis. The myelocytes invaded through haversian and Volkmann's canals from the bone marrow to periosteal areas. Hematopoiesis was suppressed by marked proliferation of tumor cells in the bone marrow of the whole bone. Atrophy was also seen in the bones, including medullary bones of the chickens suffering from ML. Proliferation of myelocytes was seen in the bone marrow and periosteum of ossified cartilaginous rings of the trachea and larynx. Marked proliferation of myelocytes was seen in the dura mater of spinal cords, and it subsequently depressed the spinal cords. Bone formation with cartilage was seen in the periosteum of the sternum having marked proliferation of myelocytes in the bone marrow and periosteum. Ultrastructurally, tumor cells showed large nuclei and cytoplasm with large round electron-dense lysosomes. The virus particles were rarely detected in the cytoplasm of tumor cells. The polymerase chain reaction test of tumor samples showed positive for subgroup J avian leukosis virus. This study indicates that the myelocytes can invade through the compact bones to the periosteum in the sternum, ribs, vertebrae, synsarcum, and ossified cartilage of trachea and larynx having thinner compact bones. In addition, the periosteal osteogenesis with cartilage in the sternum may be reactive change against the bone atrophy because of the marked proliferation of myelocytes.     

Immunohistochemical localization of neuropeptides in bovine pancreas

The occurrence and density of distribution of nerves and endocrine cells that are immunoreactive for neuropeptides in the bovine pancreas were studied by immunohistochemistry. The six neuropeptides localized were galanin (GAL), substance P (SP), methionine-enkephalin (MENK), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP). The exocrine pancreas was shown to have an appreciable number of GAL- and SP-immunoreactive nerve fibres but few fibres showing immunoreactivity for VIP and CGRP. Numerous MENK-, GAL-, SP-, and NPY-immunoreactive nerve fibres were seen in the endocrine portion of the pancreas. Nerve cell bodies in the intrapancreatic ganglia showed immunoreactivity for all of the neuropeptides except CGRP. Endocrine cells showing immunoreactivity for GAL and SP were observed in the large islets and islets of Langerhans, respectively. The present results indicate a characteristic distribution of neuropeptides in the bovine pancreas, which may regulate both exocrine and endocrine secretions of pancreas.     

Bone marrow contamination of canine cerebrospinal fluid

Bone marrow cells were identified in cytocentrifuge preparations of cerebrospinal fluid (CSF) obtained by lumbar punctures from two dogs. CSF analyses were characterized by normal or increased total cell counts, normal or increased total protein concentration and the presence of erythrocytes. Hematopoietic cells present included both erythroid and myeloid precursors and, in one case, an erythroblastic island was seen. Peripheral blood smear examination confirmed that the hematopoietic cells observed were not the result of blood contamination. Neither case was associated with a difficult tap procedure, nor with a specific disease process. Contamination may have resulted from actual marrow penetration or from extramedullary hematopoiesis in the dura mater. Accurate interpretation of CSF cytology requires the consideration of possible bone marrow contamination when hematopoietic cells are observed.     

Peripheral nerve injury associated with fracture or fracture-dislocation of the pelvis in dogs and cats: 34 cases (1978-1982)

Peripheral nerve injury was associated with fracture or fracture-dislocation of the pelvis in 23 dogs and 11 cats. In most cases, peripheral nerve injury resulted from ilial fracture with craniomedial displacement of bone fragments, or from sacroiliac fracture-dislocation with cranial displacement of the ilium. Sciatic nerve injury was associated with fracture of the acetabulum in 2 dogs. A combination of sensory, voluntary motor, and reflex abnormalities were observed in the ipsilateral extremity of affected animals. Signs of severe pain were noticed in 4 of the dogs and in 1 cat. Eight of the animals died or were euthanatized for reasons not directly related to the severity of peripheral nerve injury. Of the remaining 26 animals, 21 (81%) had good or excellent limb function 16 weeks after peripheral nerve injury was sustained. Limb function was lost or self-mutilation occurred in 4 (15%) of the animals.     

一株鸡传染性贫血病毒对不同日龄SPF鸡的致病性研究

为评价鸡传染性贫血病毒AV1550株的致病性,取1日龄、7日龄和14日龄SPF鸡分别经胸部肌肉注射不同病毒含量的病毒液,同时设置正常对照,隔离饲养观察21日。感染后14日采血测定红细胞压积,21日统计死亡率、体重变化以及胸腺、骨髓、法氏囊病变情况并测定1日龄SPF鸡感染后不同组织中的病毒载量。结果表明,1日龄SPF鸡感染AV1550株后,表现出精神沉郁、增重减缓、贫血等明显的临床症状,死亡率为53.9%;死亡鸡或观察期结束时存活鸡剖检,可见胸腺萎缩,骨髓变成淡黄色;不同剂量感染后14日,均能引起红细胞压积显著下降;21日时,胸腺病毒载量最高,可达106.7copies/mg 。7日龄SPF鸡感染后,出现增重减缓,高感染剂量（100000EID50）出现贫血,部分鸡出现胸腺萎缩和骨髓病变,但病变率低于30%。14日龄SPF鸡感染后,不引起明显临床症状。研究证实,CAV对SPF鸡的致病性具有明显的日龄依赖性,红细胞压积降低、骨髓病变、胸腺萎缩以及胸腺病毒载量测定可作为评价CAV致病的指标。     

Systemic spindle-cell proliferative disease in broiler chickens

The major organs and tissues of 24 broiler chickens (70 or 71 days old) suspected of spindle-cell proliferative disease (SPD) because of showing the tumorous lesions distributed throughout the body at meat inspection were collected for histopathological and immunohistochemical examination. Macroscopically, liver, spleen and cecal tonsil showed severe enlargement and white nodules or plaques were observed in the liver, spleen, kidney, intestine and bone marrow of the femur. All chickens were diagnosed with SPD based on the histopathological examination. The lesions of SPD were observed in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, gizzard, duodenum, jejunum, ileum, rectum, cecal tonsil, bursa of Fabricius, bone marrow of the femur and skin. Hemangioma was observed in the lung of 1 bird. Eight 1-day-old specific pathogen-free chicks were inoculated intraperitoneally with 0.25 ml of a 20% homogenate of the affected spleens of three naturally occurring cases. One inoculated bird, necropsied at 10 weeks of age, macroscopically had a white nodule in the kidney and histopathologically had spindle-cell proliferative lesions, a pattern similar to that seen in the naturally occurring cases, in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, duodenum, cecal tonsil and bone marrow of the femur, and was diagnosed with SPD. Immunohistochemically, significant positive reactions with a rabbit antiserum against avian leukosis virus antigens were detected in all spindle cells in the proliferative lesions of all examined SPD cases and in tumor cells of the hemangioma of a field case.     

Immunocytochemical detection of synaptophysin in enteric neurones during prenatal development in the rat stomach

Summary In this study, the localization and appearance of synaptophysin-immunoreactive (IR) nerve cells and their relationships with the developing gastric layers were studied by immunocytochemistry and light microscopy in the embryonic rat stomach. The stomachs of Wistar rat embryos aged 13-21 days were used. The first neuronal bodies and their processes containing synaptophysin-immunoreactivity were observed on embryonic day 13. In contrast, synaptophysin-IR nerve terminals were first observed between mesenchymal cells on embryonic day 14. These results indicate that synaptophysin is expressed in growing neurits and neuronal cell bodies before these neurones have established synaptic connections. The occurrences of mesenchymal cell condensation near synaptophysin-IR neuroblasts on embryonic day 15 reflect an active nerve element-specific mesenchymal cell induction resulting in the morphogenesis of muscle cells. Similarly, the appearance of glandular structures after synaptophysin-IR neuroblasts, on embryonic day 18, suggests that the epithelial differentiation may be closely related to the neuronal maturation as well as other factors. Finally, synaptophysin is functionally important in neuronal development and maturation, together with the establishment of neuroneuronal and neuromuscular contacts and in epithelial differentiation.     

Distribution of nitric oxide synthase (NOS) immunoreactivity in the chicken stomach

NOS-immunoactive nerve structures were studied in the domestic hen (Gallus domesticus) stomach using single-labelling immunofluorescence. The examined tissues were collected from 10-12 months old laying hens (n=11), Isabrown and Zielononózka races. The distribution and number of the nerve fibres was examined considering differences between the proventriculus and gizzard, and also between particular layers of the stomach wall. Large or moderate numbers of nerve fibres showing NOS-immunoreactivity (NOS-IR) were found in all major layers of both parts of the stomach. In the muscular layer of the gizzard, NOS-positive nerve cell bodies were also observed. Small or moderate numbers of nerve terminals were distributed around blood vessels. NOS-IR nerve fibres associated with blood vessels were slightly more numerous in the gizzard than in the proventriculus. The widespread occurrence of NOS in both parts of the hen stomach can prove its important role in the control of digestive processes. It seems that there is a reasonable necessity of further, more detailed studies on morphological characteristics of the nitrergic nerve structures the bird alimentary tract, their functional significance and plasticity in different physiological and pathological conditions.     

Mouse model of sublethal and lethal intraperitoneal glanders (Burkholderia mallei)

Sixty male BALB/c mice were inoculated intraperitoneally with either a sublethal or a lethal dose of Burkholderia mallei China 7 strain, then killed at multiple time points postinoculation. Histopathologic changes were qualitatively similar in both groups and consisted of pyogranulomatous inflammation. In sublethal study mice, changes were first seen at 6 hours in mediastinal lymph nodes, then in spleen, liver, peripheral lymph nodes, and bone marrow at day 3. These changes generally reached maximal incidence and severity by day 4 but decreased by comparison in all tissues except the liver. Changes were first seen in lethal study mice also at 6 hours in mediastinal lymph nodes and in spleens. At day 1, changes were present in liver, peripheral lymph nodes, and bone marrow. The incidence and severity of these changes were maximal at day 2. In contrast to sublethal study mice, the incidence and severity of the changes did not decrease through the remainder of the study. The most significant difference between the two groups was the rapid involvement of the spleen in the lethal study mice. Changes indicative of impaired vascular perfusion were more frequently seen in the sublethal study mice. Our findings indicate that mice are susceptible to B. mallei infection and may serve as an appropriate model for glanders infection in a resistant host such as human beings. Additionally, by immunoelectron microscopy, we showed the presence of type I O-antigenic polysaccharide (capsular) antigen surrounding B. mallei.     

Hemorrhagic enteritis: virus distribution and sequential development of antibody in turkeys

Turkeys poults were inoculated intraperitoneally with hemorrhagic enteritis virus (HEV) at 4-1/2 weeks of age. Antibody response and sequential development of viral antigen in various tissues were monitored. An enzyme-linked immunosorbent assay (ELISA) was developed to study antibody production, and immunoperoxidase staining was used to determined sites of localization of the viral antigens in tissues. Results of ELISA and immunodiffusion tests were compared. ELISA detected antibody from day 3 post-infection (p.i.), and gel diffusion detected antibody from day 5 p.i. Peak ELISA antibody titer appeared from day 14 p.i. HEV antigen was detected from 2-6 days p.i. in the spleen, liver, intestine, kidney, and bone marrow; peak titers in the spleen were on day 3 p.i. Virus was not detected after day 6 p.i.     

Chloramphenicol toxicosis in cats

Six cats were given chloramphenicol orally at the dose level of 120/mg/kg/day in 3 divided doses for 14 days and were then observed for another 3 weeks after treatment. Five other cats were used as untreated controls for the first 14 days and subsequently were given 60 mg of chloramphenicol/kg/day for 21 days. Clinical signs of toxicosis, which were more severe in cats given the higher dose level, included central nervous system depression, dehydration, reduced food intake, body weight loss, sporadic diarrhea, and vomiting. In cats given the higher dose level, chloramphenicol caused reversible marrow suppression, with marrow hypoplasia, maturation arrest of erythroid cells, and inhibition of mitotic activity, and caused vacuolation of lymphocytes and of early myeloid and erythroid cells. Significant changes were evident in bone marrow after treatment for 1 week and in peripheral blood at the end of the 2nd week. Hematologic changes included decreased numbers of neutrophils, lymphocytes, reticulocytes, and platelets. In cats given the lower dose level, changes in blood and bone marrow were similar but less severe.     

Systemic Mastocytosis in 16 Dogs

The clinical and pathologic features of systemic mastocytosis in 16 dogs are reported. There was no apparent breed or sex predilection, and the median age at presentation was 9.5 years. In 14 of 16 cases there was a primary cutaneous mast cell tumor (MCT). When cutaneous tumor location was compared with previous reports, there was no association between location and systemic dissemination. The most common presenting signs associated with the cutaneous tumor were regional dissemination, edema, ulceration, and abscessation. They were present in 12 dogs (69%). Signs of systemic illness, including anorexia, vomiting, and diarrhea, were seen in eight dogs (50%). Other than the cutaneous tumors, the most consistent physical and radiographic abnormalities included lymphadenopathy, splenomegaly, and hepatomegaly. Eosinophilia and basophilia were seen in two and five dogs, respectively. Six dogs had increased numbers of mast cells in peripheral blood or buffy coat smears. Five of the nine dogs evaluated had increased numbers of mast cells in bone marrow aspirates. Bone marrow aspiration was superior to both peripheral blood and buffy coat smears in predicting mastocytosis. Coagulation abnormalities were seen in three of five dogs tested. Using a conventional histomorphologic grading system, 10 of 13 (77%) tumors were classified as Grade III or undifferentiated and were overrepresented when compared with previous reports of cutaneous MCTs. Eighty-eight percent of the dogs either died or were euthanatized because of their tumors. Organs commonly involved at necropsy included lymph nodes, spleen, liver, and bone marrow; four dogs had gastroduodenal ulcers.