Bronchoalveolar immune defense in cattle exposed to primary and secondary challenge with bovine viral diarrhea virus

To evaluate immune defense mechanisms against bovine viral diarrhea virus (BVDV), four calves received primary and secondary intrabronchial infections with the cytopathic, type I Singer strain of BVDV. The cellular and humoral responses to these site-specific infections with BVDV were monitored by sequential bronchoalveolar lavages (BAL) conducted prior to infection (day 0, non-infected controls) and on days 4, 7, 10, 17 (day 31, secondary infection), 35, 38, 41, 48 and 62 post-infection. Peak quantities of BVDV were recovered from BAL on day 4. BVDV clearance from the lung was complete between days 17 and 31. Immune clearance of BVDV from the lower airways upon secondary infection was swift, within 4 days, and sustained throughout a 1-month period. Total numbers of BAL CD4(+) and CD8(+) T-lymphocytes increased >200-fold by day 10, and increased to levels >70-fold higher than background by 4 days after a secondary BVDV infection. gammadelta(+) T-lymphocytes increased 100-fold by day 7 and remained at levels at least 10-fold higher than pre-infection throughout the study. B-lymphocytes increased to levels 30-fold greater than pre-infection levels by day 10, and further increased to levels 100-fold higher following secondary BVDV infection. Activation (defined by the phenotype CD25(+)/CD62L(-)) and memory (defined by the phenotype CD45RO(+)/CD45R(-)) profiles of lymphocytes in the lower airways were characterized. Activated subpopulations of CD4(+) and CD8(+) cells increased nearly 300- and 150-fold, respectively, by day 10, and to levels 100- and 50-fold 4 days after the secondary infection. Memory subpopulations of CD4(+) and CD8(+) cells increased to levels 170- and 120-fold, respectively, by day 10, and to levels approximately 400- and 300-fold, respectively, 7 days after the secondary infection. The primary antibody response consisted of increased titers of anti-BVDV-specific IgA in bronchoalveolar lavage fluid (BALF). A strong secondary antibody response with high levels of anti-BVDV-specific IgA and IgG in BALF before day 4 post-secondary BVDV infection, likely contributed, along with cellular defenses, to the rapid clearance of virus from the lung upon secondary exposure. These results demonstrate that primary infection of the bovine lung with BVDV initiates cell-mediated immune responses capable of clearing the virus within 2-3 weeks. Furthermore, populations of immune-activated and memory T-lymphocytes, combined with BVDV-specific antibody production, contribute to rapid BVDV clearance upon secondary exposure to the virus.     

Multifocal cutaneous histiocytic sarcoma in a young dog and review of histiocytic cell immunophenotyping

A 9‐month‐old male Great Dane had progressive generalized nodular dermatopathy for several months. There were > 100 raised, alopecic, firm, painful nodules throughout the skin. Aspirates from several lesions yielded moderate numbers of irregularly round or polygonal to spindle‐shaped cells with mild to moderate anisocytosis and few inflammatory cells, and the cytologic interpretation was proliferation of mesenchymal or histiocytic cells. On histopathologic examination, nodules were composed of densely packed sheets of round to spindle‐shaped cells with mild anisokaryosis and low mitotic activity. Multifocal histiocytic sarcoma with a spindle‐cell pattern was diagnosed based on morphologic features and intense expression of CD18. Additional immunophenotypic analysis on frozen sections of tissue confirmed the diagnosis of histiocytic sarcoma; expression of CD18, CD45, CD1a, CD11b, and CD11c, limited expression of Thy‐1 (CD90) and CD80, and lack of expression of CD4, CD11d, and CD86 indicated that the cells were likely interstitial dendritic cells; a review of reactive and neoplastic dendritic cells is provided. Based on staging, internal organs were not affected. Sequential treatment with lomustine and doxorubicin failed to prevent progression of the cutaneous lesions, and the dog died 3 months after initial diagnosis. At necropsy, a focus of neoplastic cells was present in one lymph node, but except for skin other organs were not involved. The clinical presentation of histiocytic sarcoma may be unusual, and neoplastic cells may lack overt features of malignancy on cytologic and histopathologic examination. In some instances, immunophenotyping is required to differentiate histiocytic sarcoma from other histiocytic disorders.     

Prospects for a novel vaccine against tuberculosis

The development of a new and improved vaccine against tuberculosis has in the last 10 years been accelerated tremendously from the completed Mycobacterium tuberculosis genome and the progress in molecular biology. This has resulted in the identification of a large number of antigens with potential in tuberculosis vaccines. The next phase of this work has now started--putting the most relevant molecules back together as fusion molecules and cocktails. This requires carefully monitoring of aspects as immunodominance, recognition in different populations as well as the influence of different adjuvants and delivery systems. The most advanced of these vaccines such as the fusion between ESAT6 and Ag85B have been evaluated in a range of animal models including non-human primates and are now entering into clinical trials. For these vaccines to be successfully implemented in future vaccination programmes it is necessary to understand the immunological background for the failure of BCG and optimize the vaccines for their ability to boost the immuno-response primed by BCG.     

Molecular evidence for the infection of zoo chimpanzees by pig Ascaris

We here describe the transmission of the pig roundworm, Ascaris suum to chimpanzees maintained in the Copenhagen Zoo, Denmark. Using a technique for whole genome fingerprinting, amplified fragment length polymorphism (AFLP) and the technique PCR restricted fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer region (ITS) of the nuclear ribosomal DNA, the worms from the chimpanzees were compared with Ascaris spp obtained from humans and pigs in order to identify the source of the infection. By the use of different distance and clustering based methods on the AFLP data set the worms from the chimpanzees were assigned to the same cluster as that of the worms from pigs. The PCR-RFLP analysis supported the AFLP results. Therefore, the zoo chimpanzees have required Ascaris infections by cross-infection from pigs. Pigs as a potential source of Ascaris infections for both captive and wild chimpanzees and other animals, therefore needs to be considered and appropriate steps taken to prevent such infection.     

In vitro drug susceptibility pattern of Mycoplasma alligatoris isolated from symptomatic American alligators (Alligator mississippiensis).

A recently described mycoplasma, Mycoplasma alligatoris, was isolated from dead American alligators (Alligator mississippiensis) that had demonstrated clinical signs of lethargy, anorexia, bilateral ocular discharge, edema. paraparesis, and polyarthritis. The in vitro minimum inhibitory concentration for nine antibacterial agents was determined through serial dilution in broth and plate culture for M. alligatoris isolates. The inhibitory concentration obtained for doxycycline, enrofloxacin, sarafloxacin, oxytetracycline, tilmicosin, and tylosin (< 1 microg/ml) was lower than that of clindamycin (1-8 microg/ml), chloramphenicol (8-16 microg/ml), and erythromycin (32-138 microg/ml).     

Oropharyngeal dysphagias in the dog: A cinefluorographic analysis of experimentally induced and spontaneously occurring swallowing disorders

Cinefluorography and videofluorography were used to record and analyze functional swallowing deficits of 12 dogs with spontaneously occurring oropharyngeal dysphagias and six experimental dogs with selected neurectomies. Ten of the 12 dogs had dysphagias affecting the cricopharyngeal stage of the oropharyngeal phase of swallowing. Two dogs had mixed oropharyngeal dysphagias. Clinical signs of cricopharyngeal dysphagia could not be differentiated from those of dysphagias due to pharyngeal or mixed oropharyngeal deficits. Signs of cricopharyngeal dysphagia consisted of: 1) repeated attempts to swallow; 2) excessive head movement; 3) dropping food from the mouth after unsuccessful swallowing attempts; 4) reingestion of dropped food. Nine of these dogs had cinefluorographic evidence of asynchrony between the normal pharyngeal contraction and relaxation, and subsequent cricopharyngeal relaxation and contraction. Only one dog demonstrated a consistent cricopharyngeal non-opening (achalasia). Seven of the dogs responded dramatically to cricopharyngeal myotomy. Two dogs with mixed oropharyngeal dysphagias had poor contractility of the pharyngeal muscles in addition to cricopharyngeal dysphagia. Clinical and cinefluorographic evaluation following cricopharyngeal myotomy of one dog verified exacerbation of functional deficits due to the iatrogenic cricopharyngeal chalasia. Esophagopharyngeal reflux accentuated the contrast medium retention in the pharynx and laryngotracheal aspiration. The need was stressed for careful differentiation between cricopharyngeal dysphagia and dysphagias involving the pharyngeal stage. Four experimental dogs with selective bilateral neurectomies of branches of the glossopharyngeal (IX) and vagus (X) nerves were evaluated clinically and cinefluorographically in an attempt to identify the pathogenesis of cricopharyngeal dysphagia. The variable results in the four dogs and the observed recovery in two dogs suggested that peripheral motor nerve deficits are not a major cause of cricopharyngeal dysphagia. Glossopharyngeal neurectomy in two dogs induced a profound functional disorder involving the pharyngeal and cricopharyngeal stages and the esophageal phase of swallowing. This would support a new hypothesis that the glossopharyngeal nerve is sensory to the esophagus as well as the pharynx, and may play a major role in disorders of the pharynx, upper esophageal sphincter, and esophagus, including congenital or acquired megaesophagus.     

The longevity of immunoglobulin preservation in canine skin utilizing Michel's fixative

Skin biopsy specimens from 7 dogs with immune-mediated skin diseases diagnosed by routine histology and 5 dogs with other skin diseases were placed in Michel's transport medium for 4 to 9 years. Direct immunofluorescence yielded positive results in tissue samples from 3 dogs with pemphigus foliaceus and 2 dogs with discoid lupus erythematosus. Direct immunofluorescence was not seen in tissue samples from 1 dog with pemphigus foliaceus and 5 dogs with non immune-mediated skin diseases. Direct immunofluorescence was seen in skin biopsy specimens maintained in Michel's medium for 4 to 8 years.     

Motility of the rumen and abomasum during hypocalcaemia.

The relationship between plasma calcium level and rumen motility in cows and sheep and abomasal motility in cows was investigated by inducing hypocalcaemia in seven cows and five sheep by the infusion of Na2 EDTA over a period of approximately two hours. Rates and amplitudes of rumen and abomasal contractions were markedly reduced by the reduction of plasma calcium level to approximately 50% of normal. There were significant positive linear relationships (P less than 0.05) between rate and amplitude of rumen contractions in both sheep and cows over a plasma calcium range of 1-3 mmol/L. There was also a significant linear relationship (P less than 0.05) between plasma calcium and abomasal rate of contraction over the same range in cows, but the relationship with amplitude of abomasal contraction was not quite significant (P less than 0.1 greater than 0.05).     

Cover Image,Volume 16, Issue 1

The cover image, by Alexandra Keller et al., is based on the Original Article The JAK2/STAT5 signaling pathway as a potential therapeutic target in canine mastocytoma, DOI: 10.1111/vco.12311