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- PublicationOpen AccessComparative performance of five recombinant and chimeric antigens in a time resolved fluorescence immunoassay for detection of Toxoplasma gondii infection in cats(Elsevier, 2022-03-28) Huertas López, Ana; Contreras Rojo, Marinela; Sukhumavasi, Woraporn; Martínez Subiela, Silvia; Álvarez García, Gema; López Ureña, Nadia María; Cerón Madrigal, José Joaquín; Martínez-Carrasco Pleite, Carlos; Sanidad Animal; Facultades de la UMU::Facultad de VeterinariaFelids are definitive hosts of Toxoplasma gondii, being the only hosts that can spread the infection through oocyst shedding in their feces. The elevated presence of this parasite in the domestic cat (Felis catus), and its close contact with humans, make it necessary to obtain reliable diagnostic methods to detect positive animals as a public health measure. For this reason, in this study, the diagnostic performance of five different recombinant antigen-based techniques was assessed to diagnose T. gondii infection in cat blood plasma samples. Specifically, four T. gondii recombinant antigens (GRA7, truncated GRA7, SAG2, and truncated SAG2) and a chimeric antigen (SAG1-GRA8) were used. A time-resolved fluorescence immunoassay (TRFIA) was developed for each antigen, and the results of each of these techniques were compared with those obtained by a commercial enzyme-linked immunoassay (ELISA) and a modified agglutination test (MAT) as reference techniques. The TRFIA based on SAG1-GRA8 antigen showed better discrimination between seropositive and seronegative cats (p < 0.001), as well as a better area under the curve (0.95), sensitivity (93.6%), and specificity (89.5%) values for the optimal cut-off, versus the other TRFIAs. In addition, SAG1-GRA8 TRFIA showed substantial agreement (kappa value =0.78) and a oderate significant correlation (Spearman’s correlation: r = 0.62, p < 0.001) compared with the reference techniques. On the other hand, since plasma samples were obtained from 101 cats in Bangkok city and four of them were Neospora caninum seropositive by indirect immunofluorescence assay (IFAT), this is the first time that anti-N. caninum antibodies are detected in cats in Thailand. In conclusion, our study highlights that the TRFIA with TgSAG1-GRA8 antigen is an accurate and recommended diagnostic technique for detecting anti-T. gondii antibodies in cats.
- PublicationOpen AccessDuodenal microanatomy of the domestic cat. (Felis catus)(Murcia : F. Hernández, 1986) Robert, Henry W.; Al-Bagdadi, F.K.Duodenal samples were taken from similar locations in six cats, processed, stained, and examined via light microscope. There were no prominent circular folds (plicae circulares) or stratum compactum (lamina subglandularis). The 1072 pm X 201 pm villi were covered by 46 pm high columnar epitheliocytes proximally which decreased in height (41 pm) distally and displayed a 1.1 - 1.7 pm striated border. Globular leukocytes, mononuclear cells, and twenty-eight goblet cells (exocrinocytus calciformis) per villus were seen. The intestinal gland (crypt of Lieberkuhn) epithelium was 20 pm tall and had a less distinct striated border. The 515 pm simple straight tubular intestinal gland layer displayed distal branching. Many mitotic figures, 12 goblet cells per gland, and occasional columnar to triangular cells with red cytoplasmic granules were seen. The thickness of the lamina propria mucosa (glandular portion) decreased from proximal to distal (563-465 pm). The lamina muscularis mucosa had two layers and decreased in thickness distally (71-28 pm). The proximal muscularis mucosa was penetrated by the ducts of submucosal (Brunner's, duodenal) glands. The tela submucosa decreased in thickness distally (593-192 pm) and contained submucosal glands with 11.5 - 75 pm lumina for the first 1.5 - 2.5 cm. However, submucosal glands could be found to a distance of 8 cm. The glandular epithelium ranged from 7.5 - 22.5 pm in height. Only one type of secretory cell was observed, with both mucous and serous properties. The tunica muscularis ranged from 190-1425 pm (median thickness of 557 pm) and had two layers.
- PublicationOpen AccessElectron microscopic study of sprouting dendrites in the ciliary ganglia of cat and monkey (Macaca fascicularís) following pre- and post-ganglionic axotomy(Murcia : F. Hernández, 1997) Zhang, Y.L.; Tan, C.K.; Wong, W. C.The present paper reports the ultrastructure of dendritic sprouting and formation of associated synapses in the ciliary ganglion of cat and monkey induced by pre- and post-ganglionic axotomy. In both series of experiments, sprouting dendrites were observed mostly at 3-5 days postoperatively; such profiles were identified by their dense packing of mitochondria and glycogen-like granules. In longitudinal section, such profiles appeared as expanded extensions from the normal-looking dendritic trunks. None were observed to arise directly from the neuronal soma. After preganglionic nerve section, the cross-sectional diameters of such profiles measured 2.211 .O pm (range: 0.9-6.2 pm) in cat and 2.4I0.7 y m (range: 0.9-5.5 pm) in monkey. After postganglionic nerve section, the crosssectional diameters of such profiles measured 2.110.7 pm (range: 0.8-4.5 pm) in cat and 2.811.4 pm (range: 1.1-7 .O p m) in monkey. After preganglionic axotomy, in both cat and monkey, the axon terminals began to degenerate at 3 days postoperatively and disappeared by 5 days postoperatively. However, at later postoperative survival periods, the axon terminals reappeared and were observed to make synaptic contacts with the sprouting dendrites. Some of the sprouting dendrites were observed to degenerate, some as early as 3 days postoperatively; such profiles did not appear to have any synapse on them. After postganglionic axotomy, such sprouting dendritic profiles were also observed to make synaptic contacts with axon terminals; some were only closely associated with profiles filled with synaptic vesicles. The results thus suggest that through the formation of new synapses, sprouting of dendrites may have a role to play in neuronal survival after axotomy.
- PublicationOpen AccessEzrin and moesin expression in canine and feline osteosarcoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Hlavaty, Juraj; Wolfesberger, Birgitt; Hauck, Marlene; Obermayer Pietsch, Barbara; Fuchs Baumgartinger, Andrea; Miller, Ingrid; Walter, IngridBiological features of canine osteosarcomas (OS) differ markedly from those found in feline and resemble more human osteosarcomas, in particular for their high rate of metastasis and poor prognosis. Ezrin, radixin and moesin are members of the ERM protein family and link the actin cytoskeleton with the cell membrane. Ezrin and moesin have been shown to be of prognostic significance in tumor progression due to their role in the metastatic process. The objective of this study was to analyze ezrin and moesin protein expression in a series of dog (n=16) and cat (n=8) osteosarcoma samples using immunohistochemistry and western blot techniques. We found that cat OS have a higher moesin expression compared to dog OS, however, the active phosphorylated forms of moesin and ezrin Tyr353 were more abundant in the dog samples. A statistically significant difference was found for the low and high immunohistochemical scores of ezrin and pan-phosphoERM proteins between cat and dog. Although phosphoezrin Thr567 was higher in feline OS, the membranous localization in dog OS samples indicates the presence of the biologically active form. Therefore, the observed differences in phosphorylated forms of ezrin and moesin status should be further studied to demonstrate if they are relevant for different biological behavior between dog and cat OS.
- PublicationOpen AccessImmunohistochemical investigation of amyloid ß-protein (Aß) in the brain of aged cats(Murcia : F. Hernández, 2005) Brellou, G.; Vlemmas, I.; Lekkas, S.; Papaioannou, N.To clarify the immunohistochemical features of amyloid deposits and cerebral amyloid angiopathy (CAA), the distribution of the amyloid ß-protein subtypes Aß40, Aß42, Aß43 and Aß precursor protein (APP) were examined in the brains of fourteen aged cats (7.5-21 year-old). Two types of plaques were detected. The first type was characterized by Aß positive antigenic material and detected in the cortical layers of the frontal and parietal lobes of all examined cats. The second type was characterized by diffuse positive immune staining representing diffuse plaques, which were detected only in the very aged cats (17-21 years old) and distributed throughout the cortical layers of the parietal lobes. Vascular amyloid and the amyloid deposits were strongly positive-stained with the antibody Aß42. APP was exhibited in neurons and axons while the staining was stronger in the very aged cats (17-21 years old). Our findings suggest that the feline forms a spontaneous model for understanding the early changes of normal brain aging and the early stage of amyloid ‚-protein deposition.
- PublicationOpen AccessMyelinated Herring bodies in the median eminence of the cat(Murcia : F. Hernández, 1991) Pastor, F.E.; Blazquez, J.L.; Toranzo, D.; Pelaez, B.; Sanchez, A.; Alvarez-Morujo, A.J.; Amat, P.An electron-microscopic study was carried out on the median eminence of cats during post-natal development. From the moment of birth (observations performed 12 hours later) Herring bodies were seen in the fibrillary layer of the median eminence. At 45 days after birth, myelinated nerve fibres could be observed, some of them containing neurosecretory granules. The number of myelinated fibres in the median eminence increased with age and at 90 days some Herring bodies appeared surrounded by myelin sheaths; these mainly contained neurosecretory granules and a few mitochondria.
- PublicationOpen AccessPathological findings in a cat with cryptococcosis and feline immunodeficience virus infection(Murcia : F. Hernández, 1994) Ramos-Vara, J.A.; Ferrer, L.; Visa, J.This report describes the gross, histopathological, imm~~nocytochemi c aaln d electron rnicroscopic findings in a cat with systemic cryptococcosis and feline immunodeficiency virus (FIV) infection. Lymphadenopathy and cloudiness of leptomeninges were the major gross findings. Numerous cryptococcal yeasts were found in lymph nodes, brain, and lung, and were less common in the kidney and the eye. The inflammatory reaction varied i n intensity and cell type (mononuclear through granulomatous) depending on the organ involved. Yeasts were mainly within phagocytes as revealed by electron microscopy. Some inflammatory cells were imrnunocytochemicaIly-stained with anti-CD3 antibodies.
- PublicationOpen AccessSeroprevalence of Toxoplasma gondii in outdoor dogs and cats in Bangkok, Thailand(Cambridge University Press, 2021-03-10) Huertas López, Ana; Sukhumavasi, Woraporn; Álvarez-García, Gema; Martínez-Subiela, Silvia; Cano-Terriza, David; Almería, Sonia; Dubey, Jitender P.; García-Bocanegra, Ignacio; Cerón Madrigal, José Joaquín; Martínez-Carrasco Pleite, Carlos; Sanidad Animal; Facultad de VeterinariaThe aim of this study was to estimate the seroprevalence and risk factors associated with Toxoplasma gondii exposure in dogs and cats from Bangkok, Thailand. Blood samples from 318 dogs and 321 cats were tested for T. gondii antibodies by modified agglutination test (cut-off 1:25). Additionally, 18 dogs and 20 cats were longitudinally sampled for T. gondii antibodies during the same study period, between June and July 2019. The overall seroprevalence in dogs and cats was 7.9% (25/318; 95% CI 4.9–10.8%) and 18.7% (95% CI 14.4–23.0%), respectively. For dogs, risk factors identified were being a mixed-breed animal and living totally outdoors, while increasing age was shown to be a risk factor for cats. Seroconversion was not detected and titres from positive animals remained constant over longitudinal study. The present study indicates that there is a prominent presence of T. gondii in urban and peri-urban areas of Bangkok, suggesting that outdoor dogs and cats should be considered as a possible risk factor for humans.