P‐20 Clinical,cytopathological and histopathological evaluation of sporotrichosis in experimentally infected cats

The purpose of this study was to evaluate the clinicopathological aspects of experimental sporotrichosis in cats and compare the sensitivity of cytopathology, histopathology and culture as diagnostic tools in different phases of the infection. Twenty adult, mixed‐breed cats (10 males and 10 females) were inoculated subcutaneously with 10⁶ fungal microorganisms. Clinical examination was performed weekly. Cytopathologic, histopathologic and culture examinations were performed at 15, 30 and 60 days postinoculation. Culture of multiple organs was performed after euthanasia at 30 (10 cats) and 60 (10 cats) days postinoculation. Friedman parametric and nonparametric statistical analysis were applied to the results. The nodular, tumoral and necrotic lesionsprogressed significantly until day 30 postinoculation, and partial spontaneous regression occurred at day 60, particularly in males. An intense inflammatory pyogranulatomous and lymphocytic infiltrate with rare giant cells and sparse fibrosis associated with numerous, pleomorphic, intra‐ and extracellular fungal cells were observed on day 30. These findings gradually decreased by day 60. Despite the inflammatory granuloma associated with feline sporotrichosis, a tendency for dissemination was observed, with fungal isolation in the lymph nodes, spleen and liver at the 30 and 60 days postinoculation. No significant differences were observed between cytopathology, histopathology and fungal culture during the different phases of the disease. Therefore, cytological examination was considered a simple, rapid and inexpensive diagnostic method at all stages of this disease. Funding: Self‐funded.     

Physiological Basis of Irrigation Scheduling for Seed Production in Egyptian Clover Syn. Berseem (Trifolium alexandrinum L.) Crop

A field experiment was conducted to find out the critical physiological stages of irrigation schedules inducing better growth, physiological efficiency and seed yield potential of berseem ( Trifolium alexandrinum L., Var. S-99-1). For this purpose eight irrigation treatments were made comprised of four treatments of three irrigation (W₁, W₂, W₃ and W₄), three treatments of four irrigation (W₅, W₇ and W₈) and one treatment of five irrigation (W₆) at various physiological stages i. e. regeneration, flower initiation, full bloom, seed initiation and advance seed development stage.
Thus based on the experimental results the physiological role of watering in berseem seed production could be discussed as:
With-holding of irrigation either at regeneration or at full bloom stage developed potential water stress in plants as indicated by high proline content of irrigation treatments — W₄, W₁ and W₇; and further brought out disturbance on the formation of carotene, synthesis of water soluble sugar and translocation of sugar towards reproductive organs during grain development stage. These stresses adversely affected the plant growth and flowering behaviour. The irrigation at seed initiation stage increased the seed yield. Continuous irrigation did not appear to be useful. Thus it can be concluded that irrigation at three critical physiological stages i. e. regeneration, full bloom and seed initiation was found to be essential for obtaining potential seed yield of berseem.     

Evaluation of Five Most Commonly Grown Cotton Cultivars (Gossypium hirsutum L.) Under Mediterranean Conditions: Productivity and Fibre Quality

Studies were conducted in four regions of central Greece (Larisa, Karditsa, Livadia, Thiva), to evaluate the productivity and fibre properties (fibre strength and fibre length) of the five most commonly grown cotton cultivars in these regions. The experiments lasted for 6 years (1995–2000). All the characteristics tested were strongly influenced by the factors ‘year’, ‘cultivar’ and ‘region’. Significant cultivar × region, year × cultivar, region × year and cultivar × region × year interactions occurred for all the characteristics studied. The cultivar Alegria had the highest seed‐cotton and lint yield, while Aria had the lowest ones. Both cultivars gave the highest mean lint percentage but the lowest mean boll weight (MBW). Alegria had the lowest fibre strength, while Aria had the highest. The Acala‐type cultivars, Acala SJ2 and Zeta 2, had intermediate seed‐cotton and lint yield, while Vered 171 had high seed‐cotton and satisfactory lint yield. The three cultivars mentioned above had the highest MBW and very good fibre strength. Positive correlation was observed between fibre strength and length. In spite of the fact that mean fibre length was statistically the same in all cultivars, it was influenced by the factor ‘year’, as a result of different weather conditions. The year 1996 was the most unfavourable one for seed‐cotton yield and MBW, because it was characterized by lower than average air temperatures and rather dry conditions for all study sites. The factor ‘region’, a combination of different climatic conditions and cultural practices, influenced significantly all the parameters studied.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

A novel approach to studying the effects of temperature on soil biogeochemistry using a thermal gradient bar

The temperature dependence of chemical reaction rates and microbial metabolism mean that temperature is a key factor regulating soil trace gas emissions and hydrochemistry. Here we evaluated a novel approach for studying the thermal response of soils, by examining the effects of temperature on gas emissions and hydrochemistry in (a) peat and (b) soil from a Sitka spruce plantation. A thermal gradient was applied along an aluminium bar, allowing soil to be incubated contemporaneously from 2 to 18 °C. The approach demonstrated clear differences in the biogeochemical responses of the two soil types to warming. The peat showed no significant emission of CH₄ at temperatures below 6 °C, while above 6 °C, a marked increase in the rate of release was apparent up to 15 °C (Q₁₀ = 2.5) with emissions being similar between 15 and 18 °C. Conversely, CH₄ emissions from the forest soil did not respond to warming. Nitrate availability in the peat decreased by 90% between 2 and 18 °C (P < 0.01), whereas concentrations in the forest soil did not respond. Sulphate availability in the peat decreased significantly with warming (60%, P < 0.01), while the forest soil showed the opposite response (a 30% increase, P < 0.01). Conventionally, thermal responses are studied by incubating individual soil samples at different temperatures, involving lengthy preparation and facilities to incubate samples at different temperatures simultaneously. Data collected on a given thermal response is usually limited and thus interpolated or extrapolated. The thermal gradient method overcomes these problems, is simple and flexible, and can be adapted for a wide range of sample types (not confined to soil). Such apparatus may prove useful in the optimization of management practices to mitigate the effects of climate change, as thermal responses will differ depending on land use and soil type.     

Pasteurella haemolytica A1 and Bovine Respiratory Disease: Pathogenesis

The severe fibrinonecrotic pneumonia associated with pneumonic pasteurellosis usually results from colonization of the lower respiratory tract by Pasteurella haemolytica biotype A, serotype 1(A1). Despite recent research efforts, the authors lack a detailed understanding of the interactions and host response to P. haemolytica in the respiratory tract. The authors hypothesize that management and environmental stress factors or viral infection alters the upper respiratory tract (URT) epithelium allowing P. haemolytica to colonize the epithelium. Once the URT is colonized, large numbers of organisms enter the lung where they interact with alveolar macrophages. Endotoxin, released from the bacteria, crosses the alveolar wall where it activates pulmonary intravascular macrophages, endothelium, neutrophils, lymphocytes, platelets, complement, and Hageman factor leading to complex interactions of cells and mediators. It is the progression of this inflammatory response with neutrophil influx that is ultimately responsible for the pulmonary injury. Leukotoxin is a major virulence factor of P. haemolytica that allows it to survive by destroying phagocytic cells. At subcytolytic concentrations it may also enhance the inflammatory response by activating cells to produce mediators and release reactive oxygen metabolites and proteases.     

Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.     

Specificity of the carbon immunoassay (CIA) test for the diagnosis of Toxoplasma infections.

A rapid and low cost procedure, the carbon immunoassay (CIA) test, was evaluated for the diagnosis of Toxoplasma gondii infections. Using a closely related parasite (Besnoitia jellisoni) as antigen, and homologous or heterologous immune sera, it was demonstrated by light and electron microscopy that CIA is a very reliable and specific test. As it is neither expensive nor time-consuming, it can be recommended for general and routine laboratory use.     

Oxidative damage to the membrane of canine erythrocytes with inherited high Na, K-ATPase activity.

Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions.