Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Nonregenerative immune‐mediated anemia associated with a diffuse large B‐cell lymphoma in a captive jaguar (Panthera onca)

An 18‐year‐old male castrated jaguar (Panthera onca) was presented with anorexia and continuous bleeding from the oral cavity after a history of fighting with the partner animal. Clinical evaluation revealed ulcerating lesions on the gingiva and hard palate and a hematoma on the tongue. Computed tomography of the head and endoscopic examination of the esophagus and stomach were unremarkable. Hematology and clinical chemistry revealed severe nonregenerative anemia, mild thrombocytopenia, and moderate azotemia. Several PCRs for feline hemotropic mycoplasmas (Mycoplasma haemofelis, M heamominutium, M turicensis), Babesia felis, and Bartonella spp., as well as an FeLV antigen test were negative. The cytologic examination of a bone marrow aspirate was consistent with ineffective erythropoiesis, most likely due to immune‐mediated destruction of the erythroid precursor cells. Prednisolone therapy was initiated (1.25 mg/kg/day), and the CBC returned to normal 16 days after the initiation of the therapy. Anemia relapsed after 4 months and severe splenomegaly was noted. A repeat bone marrow aspirate revealed active erythropoiesis in the presence of erythroid precursor phagocytosis suggesting an immune‐mediated process. Splenic fine‐needle aspiration and tissue biopsies were taken, and all findings including histology and immunohistochemistry were consistent with a diffuse large B‐cell lymphoma (DLBCL). Five days later, the clinical condition deteriorated and the jaguar died. Histopathology following necropsy showed infiltration with neoplastic lymphoblasts in the spleen, liver, and abdominal lymph nodes. This case report describes a nonregenerative immune‐mediated anemia associated with a DLBCL in a jaguar.     

Antimicrobial Resistance Genes in Pigeons from Public Parks in Costa Rica

Antimicrobial resistance is known to be an emerging problem, but the extent of the issue remains incomplete. The aim of this study was to determine the presence or absence of nine resistance genes (bla_TEM, catI, mecA, qnrS, sulI, sulII, tet(A), tet(Q), vanA) in the faeces of 141 pigeons from four urban parks in Alajuela, Guadalupe, Tres Ríos and San José in Costa Rica. The genes were identified by real‐time PCR directly from enema samples. About 30% of the samples were positive for genes catI and sulI; between 13% and 17% were positive for qnrS, sulII, tet(A) and tet(Q); and 4% were positive for bla_TEM. The mecA and vanA genes were not detected. The average of antimicrobial resistance genes detected per pigeon was 2. Eight different patterns of resistance were identified, without differences in the sampling areas, being the most common pattern 2 (sulII positive samples). During rainy season, the genes more frequently found were sulI and tet(A). In conclusion, the urban inhabiting pigeons tested are currently carrying antimicrobial resistance genes, potentially acting as reservoirs of resistant bacteria and vectors to humans. To the authors’ knowledge, this is the first study carried out on direct detection of resistance genes in the digestive metagenomes of pigeons.