Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures

Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold‐free three‐dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19‐day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14‐ and 19‐day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki‐67 immunoreactivity showed an even distribution in two‐dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis‐associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold‐free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell–cell and cell–matrix interactions.     

Influence of pond fertilization and feeding rate on growth performance, economic returns and water quality in a small-scale cage-cum-pond integrated system for production of Nile tilapia (Oreochromis niloticus L.)

The effects of pond fertilization and feeding rate on growth, economic returns and water quality were investigated to develop a low‐cost cage‐cum‐pond integrated system for production of Oreochromis niloticus (L.). Hand‐sexed male fingerlings averaging 19±0.39 and 32±0.69 g were stocked in cages and open ponds at 150 fish cage^?1 and 2 fish m^?2 respectively. Fish were cultured for 114 days in five triplicate treatments. Cages were installed into ponds and caged fish were fed a 24% protein diet at 3% (T1) and 6% (T2) body weight day^?1 (BWD) without pond fertilization, and 6% BWD with pond fertilization (T3). The open water in the fourth treatment (T4) was not stocked but contained caged fish, which were fed 6% BWD for the first 57 days followed by 3% BWD for the remaining period. Ponds in the control (T5) had no cages and were neither fertilized nor open‐pond fish fed. Feeding rate and pond fertilization significantly (P<0.05) affected fish growth, profitability and water quality among treatments. Fish growth, feed utilization, fish yield, water quality and profits were significantly (P<0.05) better in T3 than the other treatments. It was concluded that fish production and economic returns were optimized at 6% BWD in fertilized ponds.     

Infection of a canine macrophage cell line with leishmania infantum: determination of nitric oxide production and anti-leishmanial activity

We have previously shown that resistance to Leishmania infantum in dogs is associated with a Th1 type of immune response. In this study, we use a canine macrophage cell line (030-D) that can readily be infected with this protozoan parasite. Our aim is to further characterize the effector mechanisms involved in killing of Leishmania parasite in dogs. We observed that activation of 030-D cells by incubation with a supernatant derived from a Leishmania-specific T cell line containing IFN-gamma, TNF-alpha and interleukin-2 (IL-2) resulted in enhanced nitric oxide (NO) production by these cells. In addition, we observed enhanced anti-leishmanial activity of infected 030-cells after activation. Both, NO production and anti-leishmanial activity were abrogated by addition of L-N(G)-nitroargininemethyl ester (L-NAME), an analogue of L-arginine. Thus, NO play an important role in the anti-leishmanial activity of these canine macrophages. We propose the infection of the 030-D cell line as a good in vitro model to further investigate parasite-host cell interactions in dogs, a natural host of Leishmania parasites.     

The effect of dialkylamine compounds and related derivatives of 1-methylcyclopropene in counteracting ethylene responses in banana fruit

Compounds that can block the ethylene receptor and be applied either as a gas or as a salt by spray or dip have been prepared and tested. Cyclopropenes with a methyl group in the 1-position, on which was attached a substituted amine, were allowed to evaporate in the presence of bananas that were treated with the gas. The minimum amount of a given compound required to inhibit chlorophyll degradation in the banana peel (an indicator of protective effect of the compound against ethylene action) that was subsequently exposed to ethylene, varied considerably depending on the compound, but N,N-dipropyl-(1-cyclopropenylmethyl)amine and N,N-di-(1-cyclopropenylmethyl)amine were the most effective. The degree of response to the ethylene inhibitory effect was similar for all of the compounds tested (32–34 d). The amount of cyclopropene compound required for inhibiting ethylene action following a 24 h exposure of bananas to the salt followed by a 15 h exposure to ethylene was higher than that required by the gas form used under the same conditions for the same effect. However, time of exposure could be much longer than 24 h with the salt than with the gas. The bananas treated with the salt do not need to be in an air-tight container, but could be used in open spaces. Only the banana peel appeared to be protected against ethylene during the 24 h interval when the salt was used. The pulp ripened upon exposure to ethylene.     

Auch ein Beitrag zur Naturgeschichte des Maulwurfs

Ohne Zusammenfassung     

The relative influence of catchment and site variables on fish and macroinvertebrate richness in cerrado biome streams

Landscape and site-scale data analyses aid the interpretation of biological data and thereby help us develop more cost-effective natural resource management strategies. Our study focused on environmental influences on stream assemblages and we evaluated how three classes of environmental variables (geophysical landscape, land use and cover, and site habitat), influence fish and macroinvertebrate assemblage richness in the Brazilian Cerrado biome. We analyzed our data through use of multiple linear regression (MLR) models using the three classes of predictor variables alone and in combination. The four MLR models explained dissimilar amounts of benthic macroinvertebrate taxa richness (geophysical landscape R ² ≈ 35 %, land use and cover R ² ≈ 28 %, site habitat R ² ≈ 36 %, and combined R ² ≈ 51 %). For fish assemblages, geophysical landscape, land use and cover, site habitat, and combined models explained R ² ≈ 28 %, R ² ≈ 10 %, R ² ≈ 31 %, and R ² ≈ 47 % of the variability in fish species richness, respectively. We conclude that (1) environmental variables differed in the degree to which they explain assemblage richness, (2) the amounts of variance in assemblage richness explained by geophysical landscape and site habitat were similar, (3) the variables explained more variability in macroinvertebrate taxa richness than in fish species richness, and (4) all three classes of environmental variables studied were useful for explaining assemblage richness in Cerrado headwater streams. These results help us to understand the drivers of assemblage patterns at regional scales in tropical areas.     

Effects of partial and complete replacement of freshwater shrimp meal (Caridinea niloticus Roux) with a mixture of plant protein sources on growth performance of Nile tilapia (Oreochromis niloticus L.) in fertilized ponds

Despite the well‐documented herbivorous food habits, commercial feeds for production of Oreochromis niloticus usually contain between 7% and 15% animal protein. However, animal protein feedstuffs are expensive, hence the need to search for cost‐effective alternatives in plant‐protein sources. Such alternatives are probably more effective in semi‐intensive systems where natural pond food forms part of the diet. This study evaluated the performance of O. niloticus after feeding diets in which fresh shrimp meal (SM) was gradually replaced by a mixture of plant‐protein sources in fertilized ponds. Three isonitronegenous (24% crude protein) diets containing 12 (control), 6% and 0% SM were fed to three groups of O. niloticus in four replicates per group for 250 days. Fish were fed daily at 2% body weight and sampled monthly to monitor growth and make feed adjustments. Growth, yields, survival and feed conversion ratio were not significantly different (P>0.05) among treatments. Growth of males was double that of females, while the sex ratio was skewed towards females. Although complete substitution of SM by plant protein did not affect the growth of tilapia, production cost was reduced by 36%. In conclusion, animal protein is not required in diets for production of O. niloticus in fertilized ponds.