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.     

Availability of minerals in rendered meat and bone meal for Nile tilapia: Preliminary observations

A digestibility trial was conducted with 180 Nile tilapia (32.65 ± 4.52 g) fed a reference diet containing 350 g/kg crude protein and 4,500 kcal/kg gross energy. Five different meat and bone meals (MBMs) were evaluated. The test diets compound by 70% of reference diet and 30% of each MBM were elaborated and classified according to ash content: MBM‐430, MBM‐400, MBM‐370, MBM‐340 and MBM‐310. The apparent availability coefficient (AAC) of minerals and the apparent digestibility coefficient (ADC) of dry matter (DM) were subjected to one‐way analysis of variance (p < .05). Differences among ADC and AAC were determined by Tukey's multiple range test (p < .05). When appropriate, a polynomial regression analysis was performed. Linear correlation analyses between MBM chemical composition and the AAC and ADC were performed (p < .01). Higher values of ADC_DM, AAC_Ca and AAC_P were observed in MBM‐310, while low values were observed in MBM‐430. Quadratic effects of ash, P and Ca contents on AAC_Ca, AAC_P and ADC_DM were observed. The AAC_Ca, AAC_P and ADC_DM indicated a negative correlation with ash, P and Ca contents. MBM‐310 exhibited the highest mineral AAC, which could be used as a mineral source for Nile tilapia.     

Magnetic resonance imaging diagnosis: incomplete ossification of the humeral condyle in a German shepherd dog

Clinical signs, radiological, arthroscopic findings and magnetic resonance imaging features of a male German shepherd dog with incomplete ossification of the humeral condyle are described. The dog showed recurrent left forelimb lameness and pain on elbow palpation. In routine radiographs, the lesion was obscured. In oblique radiographs, a radiolucent line was detected, and magnetic resonance imaging clearly demonstrated a defect in the humeral condyle. However, arthroscopy showed no changes or discontinuity of the humeral cartilage. Incomplete ossification of the humeral condyle may be underrepresented in conventional, routine studies because if there is concurrent additional elbow pathology (for example fragmented coronoid process), treatment may lead to clinical improvement and further diagnostic techniques may not be performed. The benefit of magnetic resonance imaging in this case is demonstrated.     

Evaluation of endocytic capacity and NADPH-oxidase activity from armadillo (Dasypus novemcinctus) eosinophils infected with microfilariae

Endocytic activity of phagocytic cells from armadillos infected with viruses, parasites or bacteria is unknown. This report shows that eosinophils from armadillos infected with microfilaria act against these helmintic parasites but have deficiencies in their oxygen-dependent bacteriocidal mechanisms and also in endocytic capacity against yeast.     

Mygalomorph spider community of a natural reserve in a hilly system in central Argentina

Abstract The diversity, abundance, spatial distribution, and phenology of the mygalomorph spider community in the "Ernesto Tornquist" Strict Nature Reserve were analyzed in this study. Located in southwestern Buenos Aires, Argentina, the Reserve is representative of the Ventania system, which is a sigmoidal mountain belt 180 km in length. This exceptional hilly ecosystem is home for many endemic species and rich native fauna and flora. Spider abundance was sampled monthly from October 2009 to October 2010 by hand capture and pitfall traps on grassland slopes. The species recorded in the study area were: Actinopus sp.1 (Actinopodidae); Grammostola vachoni and Plesiopelma longisternale (Theraphosidae); Acanthogonatus centralis (Nemesiidae); and Mecicobothrium thorelli (Mecicobothriidae). Grammostola vachoni and Acanthogonatus centralis were the dominant species in hand capture and pitfall traps, respectively. The seasonal variation, diversity, and abundance of the mygalomorph community are analyzed and discussed here. The Mygalomorphae of the Ventania system comprises an important group of sedentary and cryptozoic spiders that seem to be highly dependent on habitat type and environmental factors.     

Note:In vitro antagonism of actinomycetes isolated from fungus-growing ants against plant pathogenic fungi

Fungus-growing ants have been found recently to be symbiotic with actinomycetes living on the ant’s cuticle; these bacteria are inhibitory to soil fungi that are detrimental to the ants’ fungus gardens. In order to investigate whether actinomycetes found on the cuticle of attine ants also had inhibitory properties against plant pathogenic fungi, we isolated 32 strains of actinomycetes from fungus-growing ants (Atta, Trachymyrmex, andCyphomyrmex), from the Mexican states of Coahuila, Nuevo León and Tamaulipas. Of the actinomycetes tested against selected plant pathogenic fungi (Alternaria solani, Aspergillus flavus, Colletotrichum lindemuthianum, Rhizoctonia solani, Sclerotium sp.) on Czapek-Dox agar medium, 13 isolates inhibited at least one of the fungi.C. lindemuthianum was inhibited by 11 actinomycetes, andRhizoctonia by three. An actinomycete strain isolated fromCyphomyrmex rimosus inhibited all five fungi tested. http://www.phytoparasitica.org posting July 30, 2008.     

Performance of beef heifers supplemented with sodium lasalocid

This study was conducted on 78 13-month-old crossbred beef heifers that weighed 215 kg in Southern Rio Grande do Sul (RS) State, Brazil. We evaluated the performance of beef heifers that were reared in a pasture system that received a mineral supplement energy-type protein with added sodium lasalocid (LAS). The heifers were randomly and uniformly divided into 2 groups, with 39 animals in each group. One group of animals received a mineral supplement energy-type protein without sodium lasalocid (CON), and the other group received a mineral supplement energy-type protein with added LAS. The mean feed intake, the body weight (BW), the average daily gain (ADG), the body condition score (BCS), and ovarian cyclicity were recorded, and economic parameters were calculated. No differences in supplement intake were observed between the groups, which ensures adequate intake of the other components of the mineral mixture, which are part of the nutritional requirements for the production process. Similarly, no difference in the ADG was observed between treatments. We observed that the heifers in the LAS group had a higher BW gain (51 kg) that the CON heifers (40 kg; P?<?0.05). In addition, LAS-supplemented heifers had a higher BCS (3.53) than CON heifers (3.38) at the end of the experiment (P?<?0.05). The heifers supplemented with LAS had a higher profitability than the CON heifers, even with the higher cost of the supplement containing LAS; this effect was due to the higher live BW at the end of the study. We concluded that the administration of a mineral supplement energy-type protein with added LAS has beneficial effects on beef heifers in terms of production and economic feasibility.     

Sheep fed with banana leaf hay reduce ruminal protozoa population

A ciliate protozoa suppression can reduce methane production increasing the energy efficiency utilization by ruminants. The physicochemical characteristics of rumen fluid and the profile of the rumen protozoa populations were evaluated for sheep fed banana leaf hay in replacement of the Cynodon dactylon cv. vaqueiro hay. A total of 30 male sheep were raised in intensive system during 15 days of adaptation and 63 days of experimental period. The animals were distributed in a completely randomized design that included six replicates of five treatments with replacement levels (0, 25, 50, 75, and 100%) of the grass vaquero for the banana leaf hay. Samples of fluid were collected directly from the rumen with sterile catheters. Color, odor, viscosity, and the methylene blue reduction potential (MBRP) were evaluated and pH estimated using a digital potentiometer. After decimal dilutions, counts of genus protozoa were performed in Sedgewick Rafter chambers. The averages of pH, MBRP, color, odor, and viscosity were not influenced by the inclusion of the banana leaf hay. However, the total number of protozoa and Entodinium spp. population significantly decreased at 75 and 100% inclusions of banana leaf hay as roughage.     

Identification and characterization of a novel etiological agent of mango malformation disease in Mexico, Fusarium mexicanum sp. nov

The primary objective of this study was to characterize Fusarium spp. associated with the economically devastating mango malformation disease (MMD) in Mexico. In all, 142 Fusarium strains were isolated from symptomatic mango inflorescences and vegetative tissues in eight geographically diverse Mexican states from 2002 through 2007. Initially, all the Mexican isolates were screened for genetic diversity using appolymerase chain reaction and random amplified polymorphic DNA markers and were grouped into seven distinct genotypes. Based on results of these analyses, evolutionary relationships and species limits of the genetically diverse MMD-associated Fusarium spp. were investigated using multilocus DNA sequence data and phylogenetic species recognition. Maximum parsimony analyses of a five-locus data set comprising 5.8 kb of aligned DNA sequence data indicated that at least nine phylogenetically distinct Fusarium spp. within the Gibberella fujikuroi species complex are associated with MMD, including one species within the African clade (Fusarium pseudocircinatum), two species within the Asian clade (F. mangiferae and F. proliferatum), and at least six species within the American clade (F. sterilihyphosum and five undescribed Fusarium spp.). Molecular phylogenetic analyses indicate that a novel genealogically exclusive lineage within the American clade was the predominant MMD associate in Mexico. This new Fusarium sp. caused MMD and could be distinguished from all other known species morphologically by the production of mostly sterile, coiled hyphae which are typically associated with sporodochial conidiophores together with unbranched or sparsely branched aerial conidiophores. Koch's postulates were completed for isolates of the new species on nucellar seedlings of mango cv. Ataulfo. This pathogen is formally described herein as F. mexicanum.