A morphologic and immunohistochemical study of the bronchus-associated lymphoid tissue of pigs naturally infected with Mycoplasma hyopneumoniae

Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.     

Fertility and sperm quality of broiler breeder males infected with subgroup J avian leukosis virus

In order to assess the effects of subgroup J avian leukosis virus (ALV-J) on semen quality, broiler breeder males were separated by ALV-J status (ALV-J positive = POS, ALV-J negative = NEG) at 44 wk of age. Of the 249 males originally placed at 1 day of age, 101 (40.6%) died by 43 wk of age. Observations of tumor expression and high mortality suggest that many of the males that died prior to 44 wk of age were infected with ALV-J. From 47 to 56 wk of age, hens were inseminated every third week with 7.5 x 10(7) sperm. Fertility and hatch data were collected by incubating eggs laid during the 2 wk postinsemination (WPI). The number of sperm that penetrated the perivitelline membrane of the ovum was determined from eggs laid on the eighth day postinsemination. Sperm mobility index (SMI) was determined at 58 and 60 wk of age from all males producing semen. Whereas SMI and sperm hole penetration measurements indicated that the sperm quality from treatments POS and NEG were similar, fertility was significantly greater in the POS treatment during the first (89.0% vs. 79.0%) and second WPI (59.3% vs. 45.0%). However, because of numerically higher hatch of fertile from the NEG group, the percentage of hatch of eggs set was similar between groups. These data suggest that ALV-J status of caged males has no influence on sperm quality or hatchability of eggs.     

Oral administration of yeast, Saccharomyces cerevisiae, enhances the cellular innate immune response of gilthead seabream (Sparus aurata L.)

The effects of including lyophilised whole yeast, Saccharomyces cerevisiae, in the diet on the seabream innate immune response were investigated. Gilthead seabream (Sparus aurata L.) specimens were fed four different diets for 4 weeks: a commercial diet as control and the same diet supplemented with 1, 5 or 10 g/kg yeast. After 1, 2 and 4 weeks, serum complement titres, as a humoral parameter, and phagocytic, respiratory burst, myeloperoxidase and natural cytotoxic activities of head-kidney leucocytes, as cellular parameters, were evaluated. The results showed that yeast supplements enhanced all the latter responses, but not the humoral response. This enhancement was dose-dependent except for the cytotoxic activity that was only stimulated by the lower dose of yeast assayed. As yeast cell walls are able to enhance the seabream cellular innate immune response, these results support the possible use of whole yeast as natural inmunostimulants in common fish diets.     

Dorsal laminectomy for caudal cervical spondylomyelopathy: postoperative recovery and long-term follow-up in 20 dogs

OBJECTIVE: To evaluate the postoperative morbidity and long-term outcome of dogs after dorsal laminectomy for caudal cervical spondylomyelopathy (CCSM). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Twenty dogs with CCSM. METHODS: Medical records of dogs treated by dorsal laminectomy for CCSM at North Carolina State University and Colorado State University between 1989 and 1999 were reviewed. Information on signalment, onset, progression and duration of clinical signs, diagnostic testing, sites of dorsal laminectomy, postoperative complications, length of hospitalization, and the ambulatory status on discharge was recorded. A minimum follow-up of 7 months was required for inclusion in the study. Neurologic status was graded (0 to 5) preoperatively, 2 days after surgery, and at the time of the study (final score). Improvement or worsening of the neurologic status was assessed by comparison of different scores for each dog. Additional follow-up information was obtained by means of a detailed telephone questionnaire directed at both the owner and referring veterinarian. RESULTS: Mean duration of clinical signs before surgery was 4.9 months. At admission, 15 dogs were ambulatory and 5 were nonambulatory. Neurologic status worsened in 70% of dogs 2 days after surgery but improved in all but 1 dog over the long term. Mean time to optimal recovery was 3.6 months. Long-term follow-up ranged from 7 months to 9 years (mean +/- SD, 3.2 +/- 2.4 years). Four dogs had confirmed recurrence; 2 other dogs may have had recurrence. CONCLUSIONS: Dorsal cervical laminectomy is an effective treatment for CCSM in those dogs with dorsal compression or multiple sites of involvement. CLINICAL RELEVANCE: Although most dogs' neurologic status transiently worsened after surgery, long-term outcome and recurrence rates were comparable to those seen with other surgical techniques for CCSM.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Transduction of Vero cells and bovine monocytes with a herpes simplex virus-1 based amplicon carrying the gene for the bovine herpesvirus-1 Circ protein

Herpes simplex virus-1 (HSV-1) based amplicon vectors are promising gene delivery vehicles because they have a large transgene capacity and can efficiently transduce many different cell types, including non-dividing cells, of various animal species. The Circ protein of bovine herpesvirus-1 (BHV-1) is a myristylated virion component of unknown function. Preliminary experiments with a circ gene deletion mutant indicated that Circ may influence the host's immune response by downregulating MHC-II expression in bovine monocytes. To get more insight into the function of Circ, amplicon vectors were constructed with various open reading frames (ORFs) under the control of the HSV-1 IE4/5 promoter: (i) the Circ ORF alone, (ii) a fusion ORF encoding an N-terminal Circ fused to the enhanced green fluorescent protein (eGFP), (iii) the eGFP ORF alone, and (iv) the Circ ORF in the inverted orientation. Upon helpervirus-free packaging into HSV-1 amplicon particles and transduction of Vero cells, both Circ alone and the Circ-eGFP fusion protein produced a punctate pattern within the cytoplasm, suggesting membrane association of the myristylated protein. In contrast, eGFP alone was evenly distributed over the cytoplasm of transduced cells. Upon infection of bovine buffy-coat cells, it was observed that cells of the monocyte lineage but not lymphocytes were transduced. Transgene expression reached a peak around 20h after transduction and lasted for at least 90h. Transduced monocytes underwent specific morphological changes, which may be attributed to Circ synthesis.     

Cervical injury following a horse kick to the head in two dogs

Two dogs were presented to North Carolina State University Veterinary Teaching Hospital following blunt trauma to the head delivered by a horse kick. On presentation, both dogs had resolving clinical signs directly related to the head trauma, but both also had compromise to their upper airway as a result of indirect injury to the soft tissues of the neck, visible on plain radiographs. One dog made a full recovery following a period of assisted ventilation. The other dog was euthanized at the request of the owner. These injuries illustrate the importance of evaluating the cervical spine and soft tissues of the neck following blunt trauma to the head.     

Two new nematodes,Paraseuratoides ophisterni gen. et sp. n. (Quimperiidae) and Philometra ophisterni sp. n. (Philometridae), from the swamp-eel Ophisternon aenigmaticum in Mexico

Two new nematodes, Paraseuratoides ophisterni gen. et sp. n. (Seuratoidea: Quimperiidae) and Philometra ophisterni sp. n. (Dracunculoidea: Philometridae) are described based on specimens recovered from the intestine and mesentery, respectively, of the swamp-eel Ophisternon aenigmaticum Rosen et Greenwood (Synbranchiformes: Synbranchidae) from a canal of the Papaloapan River in Tlacotalpan, State of Veracruz, Mexico. The genus Paraseuratoides is mot similar to Paraseuratum Johnston et Mawson, 1940, differing from it mainly in the absence of a bulbous inflation on the anterior end of the oesophagus and in the structure of the mouth (presence of 6 spines in addition to 6 oesophageal teeth). Neoquimperia Wang, Zhao, Wang et Zhang, 1979 and Wuinema Yu et Wang, 1992 are synonymised with Ezonema Boyce, 1971 and Paragendria Baylis, 1939, respectively, and Haplonema hamulatum Moulton, 1931 is considered a junior synonym of Ichthyobronema conoura Gnedina et Savina, 1930. Philometra ophisterni (only females) is mainly characterised by minute cephalic papillae, a greatly developed anterior oesophageal bulb separated from the cylindrical part of the oesophagus, anterior extension of the oesophageal gland anterior to the nerve ring, and by the character of large caudal projections. This is the first Philometra species recorded from inland fishes in Mexico.