Tissue Of Cats Research Articles

Expression of Apoptosis-Related Genes in Cat Testicular Tissue in Relation to Sperm Morphology and Seasonality-A Preliminary Study.

Simple SummaryProgrammed cell death (apoptosis) is a crucial process in spermatogenesis, responsible for the elimination of abnormal sperm cells and for the reduction of testicular volume outside the breeding season. Poor sperm morphology (teratozoospermia) and lower semen quality out of season are commonly observed in domestic cats, but the exact reasons and mechanisms are not known. The aim of this study was to use gene expression analysis to identify which apoptotic processes and pathways are expressed in the phenomenon of teratozoospermia and seasonality in the domestic cat. The results showed a higher expression of two antiapoptotic genes and one proapoptotic gene during the non-reproductive season, with no differences noted between normozoospermic and teratozoospermic cats. We hypothesize that during the non-breeding season there is a potential detrimental factor which activates a cascade of caspases, against which germ cells mount a defense by producing anti-apoptotic proteins. Further identification of this factor may help in the amelioration of semen quality of cats and improve feline breeding.Apoptosis is a crucial process in spermatogenesis, responsible for the elimination of abnormal sperm cells and testicular regression out of breeding season. The aim of this study was to assess if the expression of apoptosis-related genes in testicular tissue of domestic cats differed: (1) between normozoospermic and teratozoospermic donors, and (2) between reproductive and non-reproductive season. The expression of genes: BCL2L1, BCL2, BAX, BAD, FAS, FASLG, and caspases (CASP3, CASP8, CASP9, and CASP10) was analyzed by qRT-PCR in testicular tissue samples. During non-reproductive season significantly higher expression of two anti-apoptotic genes (BCL2L1 and BCL2) was observed. Additionally, there was a significant higher expression of CASP10 in teratozoospermic cats during non-reproductive than during reproductive season. No differences were noted between normozoospermic and teratozoospermic groups. Upregulation of some genes during the non-reproductive season indicates engagement of apoptotic mechanisms in the seasonal changes of semen quality in cats, however further studies on protein levels and analysis of changes on distinct testicular germinal layers are required. At the same time, teratozoospermia in the general population of cats seems to be not connected with dysregulation of apoptosis in the testes.

Basic fibroblast growth factor enhances proliferation and hepatocyte growth factor expression of feline mesenchymal stem cells.

IntroductionThe objective of this study is to evaluate the effect of basic fibroblast growth factor (bFGF) on the proliferation and secretion activity of feline adipose-derived mesenchymal stem cells (MSC).MethodsFeline MSC isolated from the subcutaneous adipose tissue of cats were cultured with or without bFGF.ResultsThe bFGF addition enhanced the proliferation of feline MSC to a significant great extent compared with that without bFGF, although the cell proliferation tended to increase with the bFGF concentration. In addition, adipogenic and osteogenic staining assay demonstrated that the bFGF addition allowed MSC to maintain the differentiation ability even after the proliferation. Moreover, no change in the surface markers of MSC was observed between the cultures with or without bFGF. A quantitative RT-PCR assay revealed that the HGF and TSG-6 expression significantly increased by the bFGF addition. The highest mRNA expression of MMP-2 was observed for cells cultured in 1000 ng/ml bFGF concentration.ConclusionsThe culture with bFGF is a promising way to enhance the proliferation, and HGF secretion ability of MSC as well as maintain their differentiation ability and immunophenotype nature.

Experimental Biology 2020 Meeting Abstracts.

Bone morphogenetic proteins (BMPs) are growth factors that belong to the transforming growth factor-β (TGF-β) superfamily, and till date 15 BMPs have been described. BMPs, first described for their role in bone and cartilage formation, also play a role in renal fibrosis in chronic kidney disease (CKD). There is evidence to indicate that in rodent models of CKD, administration of recombinant BMP1-3 increases renal fibrosis whereas administration of a BMP1-3-neutralizing antibody or BMP-7 antibody reduces renal fibrosis and preserves renal function. The aim of the present study was to investigate changes in gene expression in the renal cortex obtained from cats with kidney disease or calcium oxalate stone formers (CaOx) at necropsy, to identify BMPs associated with renal dysfunction in cats and potential fibrosis. At time of death the circulating levels of creatinine as well as symmetric dimethyl arginine (SDMA), both markers of kidney decline in cats, were significantly higher in cats with renal disease (n=11) or stone-forming cats (CaOx, n=12) when compared to controls (n=19). Using RNAseq in kidney tissue, we found a modest, but significant, increase in the expression of BMP-1 in cats with kidney disease (2.48 fold) and stone formers (1.72 fold), compared to controls (both p<0.01). While the increase in BMP-2 in CaOx cats was significant (1.46 fold; p<0.05 vs Con), the increase in cats with kidney disease was not (1.23 fold; NS). BMP2K, a BMP-2 inducible kinase, was significantly increased in both kidney disease (1.43 fold) and CaOX (1.46 fold) (both p<0.05). In contrast, a significant decrease in BMP4 was observed in both groups (<2.2 fold and 1.68 fold in kidney disease and CaOx, respectively; both p<0.001 vs Con). A decrease was also seen in CRIM 1, a protein associated with podocyte filtration function and whose reduction is associated with fibrosis, in both groups. BMP-7, whose potential therapeutic role in treating CKD and reversing fibrosis has been documented, was modestly decreased in both groups (both less than 1.5 fold) compared to controls. Given that there was an increase in all three forms of TGFβ (TGFβ1, TGFβ2, and TGFβ3), a potent initiator of renal fibrosis, in both groups, and a decline in BMP-7, an endogenous inhibitor of TGFβ signaling in fibrosis, compared to controls, our results profile the BMPs potentially associated with renal fibrosis in cats that may contribute to kidney dysfunction. In summary, a nutritional therapy to slow the progression of kidney dysfunction may benefit from the inclusion of dietary ingredients that attenuate renal fibrosis in cats. SUPPORT OR FUNDING INFORMATION: This study was funded by Hill's Pet Nutrition, Inc.

Increase in Interleukin‐16, a Pro‐inflammatory Cytokine, and Other Distinct Markers of Lymphocytic Activation, in Kidney Tissue of Cats with Renal Dysfunction

Inflammation plays an important role in the initiation and subsequent progression of chronic kidney disease. An increase in inflammatory markers including cytokines as well as markers of oxidative stress is associated with reduced kidney function. Inflammatory markers and the underlying mechanisms that contribute to the pathophysiology of kidney disease are not clear and likely are multifactorial. Another feature associated with renal dysfunction and inflammatory response is the formation of calcium oxalate stones formed from the crystal deposits in the tubular epithelial cells. The aim of the present study was to investigate changes in gene expression in the renal cortex obtained from cats with kidney disease or calcium oxalate stone formers (CaOx) at necropsy in order to identify novel inflammatory biomarkers associated with renal dysfunction. At the time of death the circulating levels of creatinine as well as symmetric dimethyl arginine (SDMA), both markers of kidney decline in cats, were significantly higher in cats with renal disease (n=11) or stone‐forming cats (CaOx, n=12) when compared to controls (n=19). Using RNAseq in kidney tissue, we found a significant increase in the expression of IL‐16, a T‐cell chemoattractant, in both cats with kidney disease (6.72 fold) and stone formers (7.03 fold) compared to controls (both p&lt;0.0001). There was also a significant increase in caspase recruitment domain family member 11 (CARD11), an activator of T and B‐lymphocytes and NFkB, in cats with kidney disease (9.2 fold) and stone formers (10 fold) compared to controls (p&lt;0001). However, some differences between the kidney disease and CaOx groups were also observed. In cats with kidney disease there was an increased expression of LY9 (CD299), found in T and B‐cells and thymocytes, IL2Rg, a marker of lymphocyte activation, and SPP1 (Osteopontin), expressed in activated T and B‐cells (all &gt;6‐fold) when compared to controls (p&lt;0.0001) but such increased expression was not seen in CaOx cats. In summary, while IL‐16 was increased in both groups, our results indicate important molecular differences in the pathogenesis of kidney dysfunction in cats with kidney disease and stone formers, and that distinct lymphocyte markers may be involved in the persistent decline of kidney function. Our studies suggest that the optimal nutritional therapy to slow the progression of kidney dysfunction may be different for cats with kidney disease or stone formers.Support or Funding InformationThis study was funded by Hill’s Pet Nutrition, Inc

Effect of cryopreservation on sperm DNA fragmentation and apoptosis rates in the testicular tissue of domestic cats

The objectives of the present study were to evaluate the damage caused by cryopreservation on sperm DNA and estimate the percentage of cell apoptosis in tissue after thawing. Testicles of cats were sectioned into of 0.3 cm3 and 0.5 cm3 fragments and evaluated for DNA damage using acridine orange and semi-quantitatively through histo-morphological and immunohistochemical methods (caspase-3). Other fragments were placed in cryotubes with diluent containing either 3% glycerol or 3% propanediol, and were cryopreserved. Evaluation using acridine orange indicated there was a difference with use of propanediol and glycerol on DNA damage in 0.5 cm3fragments, with the latter being more effective than the former for cryopreservation. Results from histomorphological evaluations indicated there was a greater cell integrity among germ cells that were not cryopreserved, based on criteria assessed (detachment of cells from basal membrane, retraction of seminiferous tubule epithelium, visibility of the spermatogonia nucleoli and nuclear spermatogonia condensation), for both sizes of fragments. The values for these variables decreased after cryopreservation, with there being no differences as a result of size of fragment stored or between cryoprotectants used (P > 0.05). The staining for caspase-3 differed for the cytoplasm, nuclei and germ cells. Assessment of these staining patterns indicated the fresh fragments had an amount of cell damage and there was a similar amount of damage detected in cryopreserved fragments. This finding indicated that there was considerable efficacy in preserving the tissue fragments with use of the freezing protocols that were evaluated in this study.

Poly(3-hydroxybutyrate): Promising biomaterial for bone tissue engineering.

Poly(3-hydroxybutyrate) is a natural polymer, produced by different bacteria, with good biocompatibility and biodegradability. Cardiovascular patches, scaffolds in tissue engineering and drug carriers are some of the possible biomedical applications of poly(3-hydroxybutyrate). In the past decade, many researchers examined the different physico-chemical modifications of poly(3-hydroxybutyrate) in order to improve its properties for use in the field of bone tissue engineering. Poly(3-hydroxybutyrate) composites with hydroxyapatite and bioglass are intensively tested with animal and human osteoblasts in vitro to provide information about their biocompatibility, biodegradability and osteoinductivity. Good bone regeneration was proven when poly(3-hydroxy-butyrate) patches were implanted in vivo in bone tissue of cats, minipigs and rats. This review summarizes the recent reports of in vitro and in vivo studies of pure poly(3-hydroxy-butyrate) and poly(3-hydroxybutyrate) composites with the emphasis on their bioactivity and biocompatibility with bone cells.

Adipokines secretion in feline primary adipose tissue culture in response to dietary fatty acids

BackgroundObesity in cats has been associated with alterations in adipokines including: adiponectin, interleukin-6 (IL6), and tumor necrosis factor-α (TNFα). Omega-3 polyunsaturated fatty acids have multiple beneficial effects on obesity-associated disorders, and therefore may alleviate these alterations. This study aimed to determine the effects of body condition, fat depot, troglitazone, and different fatty acids on secretion of adiponectin, IL6 and TNFα from adipose tissue of healthy cats. Subcutaneous and visceral adipose tissue samples were collected from 18 healthy intact female cats, and body condition score (Range 3–7/9) was determined. Concentrations of adiponectin were measured in mature adipocytes cultures and concentrations of IL6 and TNFα were measured in stromovascular cells cultures following treatment with control medium, troglitazone at 10 μM, eicosapentaenoic acid, arachidonic acid, or palmitic acid, at 25, 50, or 100 μM.ResultsStromovascular cells of visceral origin secreted higher concentrations of IL6 than corresponding cells of subcutaneous origin (P = 0.003). Arachidonic acid treatment at 25, 50, and 100 μM increased IL6 secretion in subcutaneous (P = 0.045, P = 0.002, and P < 0.001, respectively) and visceral (P = 0.034, P = 0.001, and P < 0.001, respectively) stromovascular cells. Eicosapentaenoic acid treatment increased TNFα secretion in subcutaneous stromovascular cells at 25, 50, and 100 μM (P = 0.002, P = 0.001, and P = 0.015, respectively) and in visceral stromovascular cells at 50 μM (P < 0.001). No significant effect on medium adiponectin concentration was observed following troglitazone treatment (P = 0.4) or fatty acids treatments at 25 (P = 0.2), 50 (P = 0.8), or 100 (P = 0.7) μM. Body condition score did not have significant effects on medium concentrations of adiponectin (P = 0.4), IL6 (P = 0.1), or TNFα (P = 0.8).ConclusionsThis study demonstrated higher basal secretion of IL6 from visceral compared to subcutaneous adipose tissue, a stimulatory effect of arachidonic acid on secretion of IL6 and a stimulatory effect of eicosapentaenoic acid on TNFα from feline adipose tissue.

Gene expressions of de novo hepatic lipogenesis in feline hepatic lipidosis.

ObjectivesThe aim of this study was to evaluate if de novo hepatic lipid synthesis contributes to fatty acid overload in the liver of cats with feline hepatic lipidosis (FHL).MethodsLipogenic gene expression of peroxisome proliferator-activated receptor-alpha (PPAR-α), peroxisome proliferator-activated receptor-gamma (PPAR-γ), fatty acid synthase (FASN) and sterol regulatory element-binding factor (SREBF1) were evaluated using quantitative RT-PCR in liver tissue of six cats with FHL and compared with the liver tissue of eight healthy cats.ResultsIn liver tissue, PPAR-α, PPAR-γ and FASN mRNA expression levels were not significantly different (P >0.12, P >0.89 and P >0.5, respectively) in the FHL group compared with the control group. SREBF1 gene expression was downregulated around 10-fold in the FHL group vs the control group (P = 0.039).Conclusions and relevanceThe downregulation of SREBF1 in the liver tissue of cats with FHL does not support the hypothesis that de novo lipogenesis in the liver is an important pathway of fatty acid accumulation in FHL.

Lymphoplasmacytic Meningoencephalitis and Neuronal Necrosis Associated With Parvoviral Infection in Cats.

Neurologic manifestations other than cerebellar hypoplasia are rarely associated with feline panleukopenia virus (FPV) infection in cats. Here the authors describe lymphoplasmacytic meningoencephalitis and neuronal necrosis in 2 cats autopsied after exhibiting ataxia and nystagmus. Gross changes consisted of cerebellar herniation through the foramen magnum, with flattening of cerebrocortical gyri and narrowing of sulci. Histologically, lymphoplasmacytic meningoencephalitis, extensive neuronal necrosis, and neuroaxonal degeneration with digestion chambers were present in the telencephalon and brain stem in both cats. Frozen brain tissue of both cats was positive for parvoviral antigen via fluorescent antibody testing, and formalin-fixed, paraffin-embedded tissue sections of brain were immunoreactive for parvovirus antigen and positive for parvoviral DNA on in situ hybridization. Frozen brain tissue from 1 case was positive for parvovirus NS1 and VP2 genes using conventional polymerase chain reaction, and subsequent DNA sequencing and phylogenetic analysis revealed that the viral strain was a FPV. Reverse transcription quantitative polymerase chain reaction on formalin-fixed, paraffin-embedded brain tissue revealed high levels of parvovirus in both cases, supporting an acute and active viral infection. Although rare, FPV infection should be considered in cases of lymphoplasmacytic meningoencephalitis and neuronal necrosis in cats.

Differentiation and multipotential characteristics of mesenchymal stem cells derived from adipose tissue of an endangered wild cat (Leopardus guigna)

Adipose derived mesenchymal stem cells (AMSCs) have been isolated and used as cell therapy in domestic cats. For wild cats, AMSCs have been reported only in black-footed cats (Felis nigripes). Feline AMSCs may be useful to treat the injuries of endangered cats in captivity and to improve cloning techniques or for derivation of induced pluripotent stem cells (iPSC). The guigna (Leopardus guigna) is an endemic and endangered feline from Chile and Argentina. The objective of this study was to characterize putative AMSCs from guigna in terms of their main biological attributes such as: growth kinetics, differentiation ability, and surface marker expression. The results were compared with cat AMSCs. AMSCs were isolated from peritoneal adipose tissue of female cats and subcutaneous tissue from a female guigna. Cells were isolated and cultured in DMEM/F12, supplemented with DMEM/F12 + FBS 10%. Migration potential, Colony-forming unit assay, mesodermal differentiation and surface marker expression (CD45, CD44, CD90, MHCI, and MHCII) were evaluated. Cat and guigna AMSCs displayed similar growth properties in culture. Both AMSCs showed mesodermal differentiation potential; in vitro homing potential and similar surface marker expression. These results demonstrate that the AMSCs from subcutaneous tissue of guigna could potentially be useful in regenerative therapy in this species and could be considered for use in cloning of this wild cat.

Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor \u03b21

BackgroundChronic kidney disease (CKD) is common in geriatric cats, and the most prevalent pathology is chronic tubulointerstitial inflammation and fibrosis. The cell type predominantly responsible for the production of extra-cellular matrix in renal fibrosis is the myofibroblast, and fibroblast to myofibroblast differentiation is probably a crucial event. The cytokine TGF-β1 is reportedly the most important regulator of myofibroblastic differentiation in other species. The aim of this study was to isolate and characterise renal fibroblasts from cadaverous kidney tissue of cats with and without CKD, and to investigate the transcriptional response to TGF-β1.ResultsCortical fibroblast cultures were successfully established from the kidney tissue of cats with normal kidney function (FCF) and cats with chronic kidney disease (CKD-FCF). Both cell types expressed the mesenchymal markers vimentin, CD44 and CD29, and were negative for the epithelial marker cytokeratin, mesangial cell marker desmin and endothelial cell marker vWF. Only CKD-FCF expressed VCAM-1, a cell marker associated with inflammation. Incubation with TGF-β1 (0–10 ng/ml) induced a concentration dependent change in cell morphology, and upregulation of myofibroblast marker gene α-SMA expression alongside collagen 1α1, fibronectin, TGF-β1 and CTGF mRNA. These changes were blocked by the TGF-β1 receptor 1 antagonist SB431542 (5 μM).ConclusionsFCF and CKD-FCF can be cultured via a simple method and represent a model for the investigation of the progression of fibrosis in feline CKD. The findings of this study suggest TGF-β1 may be involved in fibroblast-myofibroblast transition in feline CKD, as in other species.

Altered properties of feline adipose-derived mesenchymal stem cells during continuous in vitro cultivation.

Cytotherapy with mesenchymal stem cells (MSCs) has been studied in many species, and often requires in vitro cell expansion to obtain therapeutic doses of stem cells. Because the characteristics of MSCs, such as self-renewal and multi-lineage differentiation, can be altered by long-term culture, it is important to maintain stemness during cultivation. This study assessed the changes in the characteristics of feline adipose tissue-derived (fAT)-MSCs during in vitro passaging. Stem cells isolated from the adipose tissue of donor cats were cultured for seven sub-passages. Proliferation capacity was analyzed by calculating the cell doubling time and by colorimetric assay. Expression of stem cell-specific markers was evaluated by quantitative reverse transcription (qRT)-PCR and immunophenotyping. Expression of adipogenic and osteogenic differentiation markers was also measured by qRT-PCR. Histochemical staining and measurement of β-galactosidase activity were conducted to detect cellular senescence. The cell proliferation rate decreased significantly at passage 5 (P5). Gene expression levels of pluripotency markers (Sox2, Nanog and Klf4) and stem cell surface markers (CD9, CD44, CD90 and CD105) decreased during continuous culture; in most assays, statistically significant changes were observed at P5. The ability of cells to undergo adipogenic or osteogenic differentiation was inversely proportional to the number of passages. The proportion of senescent cells increased with the number of passages. These results suggest that repeated passages alter the proliferation and multipotency of fAT-MSCs. In clinical trials, early-passage cells should be used to achieve the maximum therapeutic effect.

A case of ocular infection with Onchocerca lupi in a dog from Germany.

Onchocerca lupi is an emerging zoonotic parasite infecting the ocular connective tissue of dogs, cats and humans. The only known case of canine ocular onchocerciasis in Germany was documented in 2002 in a shelter dog. However, the species of Onchocerca causing the infection could not be identified. Here, we report a case of the ocular infection with O.lupi in a dog, confirmed by PCR and sequencing of the cox1 gene. Further investigations are required to assess the risk factors for transmission and spread of the parasite in Germany.

Immunohistochemical analysis of canine and feline muscle disorders using formalin-fixed, paraffin-embedded tissues.

Histochemical techniques used in examination of muscle biopsies typically require frozen sections. Given that most of the specimens submitted to a veterinary laboratory for diagnosis are formalin-fixed, the choice of staining methods is limited. We aimed to further advance the diagnostic capabilities of pathologists presented with formalin-fixed muscle samples and to describe the differences in immunohistopathologic findings between neurogenic and myogenic muscle disorders. Based on hematoxylin and eosin staining, we defined in dogs the histologic lesions in 4 neurogenic disorders (degenerative myelopathy and polyneuropathy) and 2 myogenic disorders (dystrophin-deficient muscular dystrophy). In cats, we defined the lesions in 2 neurogenic disorders (lymphoma of nerve roots and spinal cords) and 1 myogenic disorder (laminin α2-deficient muscular dystrophy). Immunohistochemistry for slow and fast myosins revealed angular and group atrophy of type 1 and type 2 fibers in dogs and cats, and fiber type grouping in dogs. These immunohistopathologic findings were specific to neurogenic muscle disorders. Immunohistochemistry for nestin and myogenin revealed nestin-positive fibers and myogenin-positive nuclei in dogs and cats. They were not specific, but these fibers in myogenic disorders can be interpreted as regenerating fibers. The immunohistochemical method described herein appears to be useful for discriminating neurogenic and myogenic disorders in formalin-fixed, paraffin-embedded muscle tissue of dogs and cats.

Immunohistochemical evaluation of the activation of hepatic progenitor cells and their niche in feline lymphocytic cholangitis

ObjectivesThe aim of the study was to compare the hepatic progenitor cell niche in healthy feline livers and the liver tissue of cats with lymphocytic cholangitis.MethodsImmunohistochemical stainings for vimentin, laminin, beta (β)-catenin and Notch1 intracellular domain (NICD) were used on formalin-fixed liver biopsies from affected (n = 12) and unaffected cats (n = 2).ResultsAll immunohistochemical markers used were expressed in more cells, or more intensely, in the liver tissue of cats with lymphocytic cholangitis than in the liver tissue of unaffected cats.Conclusions and relevanceEnhanced expression of vimentin, laminin, cytoplasmic/nuclear β-catenin and NICD in liver biopsies from cats with lymphocytic cholangitis indicates that the hepatic progenitor cell (HPC) niche is remodelled and activated. HPCs might provide insights into new regenerative treatment options for lymphocytic cholangitis in cats in the future.

First report of feline morbillivirus in South America.

Feline morbillivirus was first identified in healthy and diseased stray cats captured in Hong Kong. Recently, it was demonstrated that the virus circulates within cat populations in Japan, Italy, Germany, and the USA. Importantly, an association between feline morbillivirus infection and chronic kidney disease was suggested by histological analysis of kidney tissue of infected cats. The aim of this study was to verify the presence and examine the genetic diversity of feline morbilliviruses associated with infections of domestic cats in Brazil. Seventeen cats without clinical manifestations of urinary tract diseases from a multi-cat household and 35 random client-owned cats admitted to the Teaching Veterinary Hospital for a variety of reasons were evaluated for paramyxoviral infection and the presence of uropathy. A fragment of the paramyxoviral L gene was amplified from urine samples using a reverse transcription semi-nested PCR assay. For the first time, we detected a feline morbillivirus strain that was genetically related to viral strains previously characterized in Japan in urine samples from cats in South America, in Brazil. This together with the recent description of feline morbillivirus identification within cat populations in the USA, suggests a possible widespread distribution of this viral agent on the American continent. Our data demonstrated feline morbillivirus RNA shedding mostly in the urine of cats without clinical, laboratorial, or ultrasonographic signs of urinary tract diseases. In contrast to previously published findings that associated feline morbillivirus infection with chronic kidney disease, we did not observe a clear relationship between feline morbillivirus RNA shedding in urine and kidney disease in the cats evaluated.

ПАТОМОРФОЛОГІЧНІ ЗМІНИ ЛЕГЕНЕВОЇ ТКАНИНИ ЗА ІНФЕКЦІЙНОГО ПЕРИТОНІТУ КОТІВ

The article presents the results of pathoanatomical, pathohistological and histochemical researches of the lung tissue for exudative form of the spontaneous infectious peritonitis of cats. The results of researches allow us to analyze pathoanatomical, microstructural changes in lung tissue of cats in various forms of peritonitis infectious and to define the mechanism of the pathological process that cause lethal consequence.The pathoanatomical research of dead bodies of 4 cats has been carried out: a 3 year–old cat and 1 year and 2 month–old cat and 1 year and 5 month–old with symptoms of exudative pleurisy; a 2 year–old cat with clear signs of exudative peritonitis and pleurisy. Peritoneal fluid and lung tissue fragments for cytological and pathohistological examination were selected. At pathoanatomical autopsy accumulation in the thoracal, abdominal cavities and pericardial space fluid with the flakes of fibrin were revealed, thickening of the serous membranes, layering them fibrin. At the pathohistological research in lung tissue noted the consequences of hemodynamic disturbances in the form of multiple perivascular hemorrhage, interstitial pneumonia, atelectasis and emphysematous cells and signs of fibrinous pleurisy in the state organization have been noted. The hardest changes were developed in the structures of the vascular system and are characterized by the damage of vascular endothelium, the development of productive meso– and periarteritis, syndrome of disseminated intravascular coagulation, stasis and hemolysis of erythrocytes in small vessels of interalveolar septum leading to the oppression of hemocirculation. In lungs, the mononuclear–macrophage infiltration prevailed, especially in periarteritis zone, indicating the presence of productively–necrotic vasculitis.The development of the syndrome disseminated intravascular coagulation led to the formation of multiple micro–blood clots, aggregated cells in vascular channel, the presence of which has led to the development of thrombi and then hemorrhages. The blockade microcirculation, in their turn, led to tissue hypoxia, acidosis and, as a result, distrophic changes in the past. The discovered processes in the vascular system of the lung tissue caused the insufficient flow of blood, irreversible changes of homeostasis and a sharp decline in adaptive capacity.

Zoonotic ocular onchocercosis caused by Onchocerca lupi in dogs in Romania

Onchocerca lupi is a filarial nematode, which infects the scleral conjunctival tissue of dogs, wolves and cats. Whilst adult nematodes localize in the conjunctive tissue of sclera or in the retrobulbar, microfilariae are found in the skin, and they are rarely diagnosed in asymptomatic animals. Since the first report of human ocular infection 5 years ago, up to 10 zoonotic cases have been identified in patients worldwide. We report, for the first time in Romania, three cases of canine ocular onchocercosis in dogs. Fragments of the harvested worms were characterized morphologically and molecularly. This article expands knowledge on the distribution of this parasite in Eastern Europe and sounds an alarm bell for ophthalmologists about the possible occurrence of human cases of O. lupi infection.

Molecular cloning of feline resistin and the expression of resistin, leptin and adiponectin in the adipose tissue of normal and obese cats.

Resistin, one of the adipokines that has a cycteine-rich C-terminus, is considered to relate to the development of insulin resistance in rats. However, in cats, there is little knowledge regarding resistin. In this study, we cloned the feline resistin cDNA from adipose tissue by RT-PCR. The feline resistin clone contained an entire open reading frame encoding 107 amino acids that had 72.8%, 75.4%, 50.9% and 51.8% homology with bovine, human, mouse and rat homologues, respectively. In both subcutaneous and visceral adipose tissues, the transcription levels of feline resistin mRNA were significantly higher in obese cats than normal cats, and those of feline adiponectin mRNA were significantly lower in obese cats than normal cats. However, there was no difference in the expression of feline leptin between normal and obese cats. On the other hand, in both normal and obese cats, there were no significant differences in resistin, leptin and adiponectin mRNA levels between subcutaneous and visceral adipose tissues. In cats, the altered expression of resistin and adiponectin mRNA with obesity may contribute to the pathogenesis of insulin resistance and subsequent diabetes mellitus. In addition to feline adiponectin, the feline resistin cDNA clone obtained in this study will be useful for further investigation of the pathogenesis of obesity in cats.

Effect of photoperiod on the feline adipose transcriptome as assessed by RNA sequencing.

BackgroundPhotoperiod is known to cause physiological changes in seasonal mammals, including changes in body weight, physical activity, reproductive status, and adipose tissue gene expression in several species. The objective of this study was to determine the effects of day length on the adipose transcriptome of cats as assessed by RNA sequencing. Ten healthy adult neutered male domestic shorthair cats were used in a randomized crossover design study. During two 12-wk periods, cats were exposed to either short days (8 hr light:16 hr dark) or long days (16 hr light:8 hr dark). Cats were fed a commercial diet to maintain baseline body weight to avoid weight-related bias. Subcutaneous adipose biopsies were collected at wk 12 of each period for RNA isolation and sequencing.ResultsA total of 578 million sequences (28.9 million/sample) were generated by Illumina sequencing. A total of 170 mRNA transcripts were differentially expressed between short day- and long day-housed cats. 89 annotated transcripts were up-regulated by short days, while 24 annotated transcripts were down-regulated by short days. Another 57 un-annotated transcripts were also different between groups. Adipose tissue of short day-housed cats had greater expression of genes involved with cell growth and differentiation (e.g., myostatin; frizzled-related protein), cell development and structure (e.g., cytokeratins), and protein processing and ubiquitination (e.g., kelch-like proteins). In contrast, short day-housed cats had decreased expression of genes involved with immune function (e.g., plasminogen activator inhibitor 1; chemokine (C-C motif) ligand 2; C-C motif chemokine 5; T-cell activators), and altered expression of genes associated with carbohydrate and lipid metabolism.ConclusionsCollectively, these gene expression changes suggest that short day housing may promote adipogenesis, minimize inflammation and oxidative stress, and alter nutrient metabolism in feline adipose tissue, even when fed to maintain body weight. Although this study has highlighted molecular mechanisms contributing to the seasonal metabolic changes observed in cats, future research that specifically targets and studies these biological pathways, and the physiological outcomes that are affected by them, is justified.