Toxicity of Haemophilus pleuropneumoniae to porcine lung macrophages

Viable Haemophilus pleuropneumoniae bacteria were toxic for porcine alveolar macrophages in vitro. This cytotoxic effect proved to be dose-related. A cell-free extract of H. pleuropneumoniae, heat-killed bacteria, and a Pasteurella multocida field strain were nontoxic. When macrophages were cultured with H. pleuropneumoniae bacteria in a ratio of 100 macrophages to six bacteria, ultrastructural signs of cellular degeneration were observed within 1 h. This degeneration was observed in macrophages with or without phagosomes containing H. pleuropneumoniae. A cytotoxic substance was filtered from a H. pleuropneumoniae culture in Eagle's minimal essential medium supplemented with Earle's salts (EMEM) and 10% foetal calf serum that was incubated for 10 h at 37 degrees C. This substance was destroyed by heating at 65 degrees C for 30 min. Macrophages were less susceptible to the toxic effect of H. pleuropneumoniae when serum of convalescent pigs was added.     

P‐33 Poxvirus infection in two cats

A 3‐year‐old Burmese cat was presented with a history of nonresolving crusted and papular lesions of the face and prior treatment with prednisolone. Skin biopsies revealed typical pox lesions with hyperplasia and ulceration of the epidermis and eosinophilic cytoplasmic inclusion bodies in the epidermal cells. In the upper dermis there was prominent diffuse mast cell infiltration and mild neutrophilic and eosinophilic inflammation. Rare cytoplasmic inclusion bodies were also present in swollen endothelial cells of dermal venules, which showed no other degenerative changes. Histological diagnosis was confirmed by electron microscopic evidence of pox virus particles in inclusion bodies of epidermal cells. The lesions resolved within 6 weeks with systemic antibiotic therapy and supportive care. A 2‐year‐old domestic short‐haired cat was presented with multiple disseminated papular and ulcerative pox lesions with central eschar over the entire body. Histologically, large epidermal inclusion bodies (up to 6 μm in diameter) were present. Widespread haemorrhage and vascular wall necrosis was visible in the dermis and subcutis. Some subcutaneous vessels showed neutrophilic vasculitis. In addition to diffuse dermal neutrophilic and eosinophilic inflammation, a lymphohistiocytic panniculitis was also present. The cat died as a result of massive haemorrhage and lymphedema, despite supportive care. Funding: Self‐funded.     

Septicaemic Escherichia coli and experimental infection of calves

Three strains of Escherichia coli with a common surface antigen, 31a, capable of adhering to calf enterocytes in vitro were compared to reference strains of septicaemic E. coli (RVC 330 and vir E. coli). The surface antigen 31a was present in the RVC 330 reference strain. E. coli vir had a surface antigen which was not present in E. coli 31a or E. coli RVC 330. The RVC 330 and vir reference strains also adhered to calf enterocytes in vitro. Oral infection of calves not receiving colostrum with E. coli 31a was generally followed by septicaemia and death in less than 48 h. Post-mortem examination revealed pneumonia and oedema of the kidneys and gall bladder. Oral infection of calves receiving colostrum had no effect, but intravenous inoculation produced arthritis within 15 days. The comparison of these results with those previously described by other workers did not lead to the identification of pathognomonic characteristics, which could be clearly correlated with properties specific to E. coli 31a. It is suggested that, like ColV and vir, antigen 31a may be a virulence marker for certain strains of bovine septicaemic E. coli. Furthermore, the 31a antigen appears to be carried on a plasmid.     

Re-defining tillage erosion: quantifying intensity–direction relationships for complex terrain

Abstract. Current tillage erosion models account for the influence of tillage direction in the magnitude of the soil (tillage) transport coefficient. It is argued here that this is counter-intuitive and causes significant problems in modelling tillage erosion in areas of complex terrain. This article examines whether a re-modelling of tillage erosion is possible that separates tillage direction (an interaction with the landform) from the soil transport coefficient (a measure of tillage intensity representing the combination of implement erosivity and soil erodibility). Experimental data for mouldboard ploughing upslope, downslope and cross-slope at Coombe Barton Farm, Devon are examined. Integration of data for all directions into a single relationship, which relates translocation in the direction of tillage to slope in the direction of tillage and translocation perpendicular to tillage to slope perpendicular to tillage, is not possible using previously published methods of analysis. However, when total translocation distance is regressed against the tangent of the slope at 45° to the tillage direction (bisecting the tillage direction and the direction of overturning) it is found that a single relationship can be used to describe tillage in all three directions. Therefore, this relationship is used to determine a single value of the soil transport coefficient ( k _fTa) for constant soil and implement conditions but different tillage directions. This redefinition of tillage is important both for true estimation of tillage erosion severity, the adirectional coefficient being 40% larger than the directional coefficient, and for modelling of tillage erosion in complex terrain. These improvements are vital when tillage erosion simulation is used to direct soil conservation strategies.     

Prevalence of Dirofilaria immitis and Dipetalonema reconditum in greyhounds

Blood samples from 331 greyhounds in the Hunter Valley and nearby coastal areas of New South Wales were examined for microfilariae using a filtration technique. Species were identified by histochemical staining; 10.9% of the greyhounds were infected with Dirofilaria immitis and 3.6% with Dipetalonema reconditum. The prevalence of infection of both species was significantly greater in summer than in winter (p = less than 0.05). Infection with D. immitis was correlated with differences in age, sex, bodyweight and coat colour, and a reported lack of stamina and the presence of a cough. No significant association was found. Diethylcarbamazine citrate was used for prophylaxis in 8.8% of all the greyhounds examined.     

Immunocytochemical study of tissues from clinically normal dogs and of neoplasms, using keratin monoclonal antibodies.

Three commonly used keratin monoclonal antibodies (MAB)--AE1:AE3, CAM 5.2, and MAK-6--were compared with routinely used cytokeratin antibody. The expression of these antibodies was analyzed in several tissues obtained from clinically normal dogs and in a variety of neoplasms from dogs. Using appropriate enzymatic digestion, paraffin-embedded tissues processed in routine manner retained their typical keratin expression. Differentiated and poorly differentiated epithelial neoplasms, lymphomas, and melanomas were studied by use of the avidin-biotin-peroxidase technique. All 4 of the aforementioned antibodies had similar staining profiles. Of 3 anaplastic carcinomas, 2 had positive reaction to all 4 antibodies. All lymphomas, plasma cell tumors, and amelanotic melanomas had negative reaction to MAK-6, CAM 5.2, AE1:AE3, and cytokeratin MAB. Three basal cell epitheliomas had positive reaction to all 4 antibodies, whereas 1 basal cell tumor with a solid pattern had negative staining reaction. Two carcinoids had negative reaction to all markers and 1 of 2 malignant chemodectomas and 1 transitional cell carcinoma had staining reaction to only AE1:AE3 MAB. Comparing the 4 antibodies, use of AE1:AE3 MAB produced the strongest staining intensity followed by cytokeratin, MAK-6, and CAM 5.2 MAB. All 4 antibodies had low background staining. In conclusion, AE1:AE3 and MAK-6 MAB are as useful as cytokeratin MAB for identification of poorly differentiated epithelial neoplasms in dogs and cats.     

Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells.

Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.