Diagnosis of feline pancreatitis with SNAP fPL and Spec fPL

Objectives Pancreatitis is a frequent disease in cats for which the ante-mortem diagnosis remains challenging. Feline pancreatic lipase immunoreactivity (fPLI) has been reported to have a high sensitivity for the diagnosis of pancreatitis. The aim of this study was to compare the rapid in-house test SNAP fPL with the standard test Spec fPL and to evaluate the use of SNAP fPL to diagnose pancreatitis in an emergency setting. Methods fPLI of 111 cats with a clinical suspicion of pancreatitis was measured with both SNAP fPL and Spec fPL. Furthermore, clinical signs, haematological and biochemical changes, and abdominal ultrasound findings were recorded. Results Seventy-eight of 111 cats (70.3%) were tested below the cut-off level for pancreatitis with SNAP, as well as Spec fPL, whereas 21/111 (18.9%) were tested with values above the cut-off level with both tests. In 12/111 (10.8%) cats the results were discordant. The comparison of both tests revealed an agreement of 78/80 (97.5%) when Spec fPL was ⩽3.5 μg/l (negative) and 18/20 (90%) when Spec fPL was ⩾5.4 μg/l (positive). The most common clinical signs in cats with suspected pancreatitis (n = 21) were lethargy (95.2%), reduced appetite and vomiting (90.5% each), dehydration (81.0%), diarrhoea (57.1%), abdominal pain and weight loss (47.6% each). Hyperglycaemia and hyperbilirubinaemia (85.7% each), increased aspartate transaminase (76.2%) and alanine transaminase (47.6%), leucocytosis (61.9%), lymphopenia (57.1%), decreased sodium and chloride (57.1% each), and increased urea (52.4%) were the most common abnormalities in blood work. Conclusions and relevance Clinical signs, as well as routine blood-work changes, were non-specific and thus proved to be insufficient to diagnose pancreatitis. The combination of SNAP fPL and subsequent Spec fPL, if indicated, provided the opportunity to rule out or to diagnose pancreatitis with a higher certainty than previously known test methods. This study proved SNAP fPL to be a valuable tool to exclude or include pancreatitis in an emergency settin

Establishment and characterization of a primary canine duodenal epithelial cell culture

Many mechanisms involved in the pathogenesis of chronic enteropathies or host-pathogen interactions in canine intestine have not been elucidated so far. Next to the clinical and in vivo research tools, an in vitro model of canine intestinal cell culture would be very helpful for studies at the cellular level. Therefore, the purpose of this study was to establish and characterize a primary canine duodenal epithelial cell culture. Neonatal duodenum was disrupted with trypsin-ethylenediaminetetraacetic acid (EDTA) and the mucosa scraped off and digested with collagenase and dispase. After centrifugation on a 2% sorbitol gradient, the cells were incubated at 37°C in OptiMEM supplemented with Primocin, epidermal growth factor, insulin, hydrocortisone, and 10% fetal calf serum (FCS). After 24h, the FCS concentration was reduced to 2.5%, and the temperature decreased to 33°C. With this method, the cultures were growing to confluent monolayers within 5-6d and remained viable for an average of 2wk. Their epithelial nature was confirmed by electron microscopy and immunofluorescence staining using antibodies directed against specific cytokeratins, desmosomes, and tight junctions. The intestinal cells proliferated, as evidenced by immunolabeling with a Ki-67 antibody, and cryptal cell subpopulations could be identified. Furthermore, alkaline phosphatase and sucrase activity were detecte

Toll-like receptors in domestic animals

Toll-like receptors are pattern recognition receptors with which hosts recognize pathogen-associated molecular patterns (PAMP). This recognition process is translated rapidly into a meaningful defense reaction. This form of innate host defense is preserved in the animal kingdom: invertebrates heavily depend on it; higher vertebrates also have an adaptive immune system. Both adaptive and innate immune systems are intertwined in that the former also depends on an intact innate recognition and response system. Members of the TLR system cover recognition of parasitic, bacterial or viral germs. Due to the constraints imposed by the necessity to recognize PAMP and to interact with downstream signaling molecules, the TLR system is relatively conserved in evolution. Nevertheless, subtle species differences have been reported for several mammalian TLR members. Examples of this will be given. In all mammalian species investigated, part of the coding sequence is available for the most important TLR members, thus allowing study of expression of these TLR members in various tissues by reverse-transcription polymerase chain reaction in its classical (RT-PCR) and quantitative real time RT-PCR (qRT-PCR) form. In some species, the whole coding sequences of the most important or even all TLR members are known. This allows construction of cDNA and transfection of common host cells, thus permitting functional studies. Extensive investigations were devoted to the study of non-synonymous single nucleotide polymorphisms. In a few cases, expression of a given amino acid in the extracellular (ligand-binding) portion of TLR members could be associated with infectious diseases. This will be discussed belo

Intestinal Tritrichomonas foetus infection in cats in Switzerland detected by in vitro cultivation and PCR

Tritrichomonas foetus, a parasite well known for its significance as venereally transmitted pathogen in cattle, has recently been identified as a cause of chronic large-bowel diarrhea in domestic cats in the US, UK, and, more recently, also in Norway. In a period of 3months (October to December 2007), 45 cats of Switzerland suffering from chronic diarrhea were investigated for intestinal infections, including a search for trichomonads. A commercially available in vitro culture system was used to screen for infection, complemented with a PCR and subsequent amplicon sequencing to support speciation. The PCR is based upon amplification of a sequence derived from the internal transcribed spacer region 1 (ITS1) on the ribosomal RNA gene (rRNA) using primers designed to detect a broad range of genera and species belonging to the family of Trichomonadidae. The method was furthermore adapted to the uracil DNA glycosylase (UDG) system in order to prevent carry-over contamination and it included a recombinant internal control to track for inhibitory reactions. Eleven out of the 45 cats were culture-positive, as revealed by microscopic identification of trichomonadid organisms. One of the isolates was subjected to scanning electron microscopy and findings revealed the presence of three flagella, thus placing the isolate into the gender Tritrichomonas sp. PCR and subsequent amplicon sequencing were carried out with ten of the 11 isolates. A total homology with published T. foetus sequences was confirmed in all of the cases. T. foetus therefore appears to range among those organisms that can cause chronic diarrhea in cats in Switzerlan

Traces of Canine Inflammatory Bowel Disease Reflected by Intestinal Organoids

Inflammatory bowel disease (IBD) is a chronic inflammatory condition that affects humans and several domestic animal species, including cats and dogs. In this study, we have analyzed duodenal organoids derived from canine IBD patients using quantitative proteomics. Our objective was to investigate whether these organoids show phenotypic traits of the disease compared with control organoids obtained from healthy donors. To this aim, IBD and control organoids were subjected to quantitative proteomics analysis via liquid chromatography–mass spectrometry. The obtained data revealed notable differences between the two groups. The IBD organoids exhibited several alterations at the levels of multiple proteins that are consistent with some known IBD alterations. The observed phenotype in the IBD organoids to some degree mirrors the corresponding intestinal condition, rendering them a compelling approach for investigating the disease and advancing drug exploration. Additionally, our study revealed similarities to some human IBD biomarkers, further emphasizing the translational and comparative value of dogs for future investigations related to the causes and treatment of IBD. Relevant proteins such as CALU, FLNA, MSN and HMGA2, which are related to intestinal diseases, were all upregulated in the IBD duodenal organoids. At the same time, other proteins such as intestinal keratins and the mucosal immunity PIGR were depleted in these IBD organoids. Based on these findings, we propose that these organoids could serve as a valuable tool for evaluating the efficacy of therapeutic interventions against canine IBD

Виробнича інтелігенція українського села: до проблеми формування (середина 1940-х – 1960-х рр.)

Paramyxoviruses constitute a large family of enveloped RNA viruses including important pathogens in veterinary and human medicine. Recently, feline paramyxoviruses, genus morbillivirus, were detected in cats from Hong Kong and Japan. Here we describe the discovery of several new feline paramyxoviruses. Infections with these diverse viruses were detected in urine samples from cats suffering from chronic kidney disease (CKD). No viral RNA was found in cats without clinical signs of uropathy highlighting an association between feline paramyxovirus (FPaV) infections and CKD. Phylogenetic analyses of the detected viruses showed that they represent at least two different species, one of them representing the feline morbilliviruses detected previously in Hong Kong and Japan. In addition, a new FPaV was detected sharing only 73 % homology on the nucleotide level of the viral L-gene to currently known paramyxoviral species

Proposal for rational antibacterials use in the diagnosis and treatment of dogs with chronic diarrhoea

Chronic diarrhoea is a frequent complaint in canine practice and the diagnostic path is often characterised by numerous diagnostic tests and stepwise empirical treatments, often applied before gastrointestinal (GI) endoscopy/mucosal biopsies. These include dietary interventions (novel protein, hydrolysed protein diet), parasiticides and still, in many cases, antibacterials. Indiscriminate use of antibacterial drugs risks detrimental consequences for both the individual patient (antimicrobial resistance, long-term disruption of intestinal bacterial populations, potential worsening of GI signs) and general public. For that reason, in this Perspective essay we advocate use of antibacterials only after histopathologic evaluation of GI biopsies or, for those cases in which endoscopy is not possible, after other therapeutic trials, such as diet/pre-probiotics or anti-inflammatory drugs have proven unsuccessful. They should be reserved, after appropriate dietary trials, for those canine chronic diarrhoeic patients with signs of true primary infection (i.e. signs of systemic inflammatory response syndrome or evidence of adherent-invasive bacteria) that justify antibacterial use

Formerly bile-farmed bears as a model of accelerated ageing

Bear bile-farming is common in East and Southeast Asia and this farming practice often results in irreversible health outcomes for the animals. We studied long-term effects of chronic bacterial and sterile hepatobiliary inflammation in 42 Asiatic black bears (Ursus thibetanus) rescued from Vietnamese bile farms. The bears were examined under anesthesia at least twice as part of essential medical interventions. All bears were diagnosed with chronic low-grade sterile or bacterial hepatobiliary inflammation along with pathologies from other systems. Our main finding was that the chronic low-grade inflammatory environment associated with bile extraction in conjunction with the suboptimal living conditions on the farms promoted and accelerated the development of age-related pathologies such as chronic kidney disease, obese sarcopenia, cardiovascular remodeling, and degenerative joint disease. Through a biomimetic approach, we identified similarities with inflammation related to premature aging in humans and found significant deviations from the healthy ursid phenotype. The pathological parallels with inflammageing and immuno-senescence induced conditions in humans suggest that bile-farmed bears may serve as animal models to investigate pathophysiology and deleterious effects of lifestyle-related diseases

Doramectin-Intoxikation bei drei Katzenwelpen

Dieser Fallbericht beschreibt drei Katzenwelpen mit Verdacht auf Doramectin-Intoxikation. Ein Wurf von insgesamt 7 Katzenwelpen wurde mit Doramectin behandelt, woraufhin drei Welpen neurologische Symptome entwickeltena Ein Welpe zeigte leichte Apathie und Zittern, ein weiterer zusätzlich Verhaltensänderungen und epileptiforme Anfälle, die mit Diazepam behandelt werden mussten. Beide Welpen erholten sich vollständig. Ein dritter Welpe wurde 3 Tage nach Doramectin-Gabe stuporös vorgestellt und zeigte im weiteren Verlauf Verhaltensänderungen wie Aggressivität, Hyperästhesie, Tremor und epileptiforme Anfälle. Er verstarb 36 Stunden nach Vorstellung. Bei der histopathologischen Untersuchung des Gehirns konnten ein zytotoxisches Ödem und Polioenzephalomalazie festgestellt werden. Die Doramectin-Dosis beim verstorbenen Welpen betrug 380 μg/kg.This case report describes 3 kittens with suspected doramectin toxicity. In a litter of 7 kittens treated with doramectin, 3 developed neurological symptoms. One kitten showed mild apathy and tremors, while a second one additionally presented behavioral changes and seizures that had to be treated with diazepam. Both kittens recovered completely. A third kitten was presented to us in coma 3 days following Treatment with doramectin. Subsequently, this kitten developed behavioral changes such as aggression, hyperesthesia, tremors, and seizures and died 36 hours after presentation. Histopathologic examination of the brain showed cytotoxic edema and polioencephalomalacia. The doramectin dosage of the deceased kitten was 380 μg/kg

Diagnosis of feline pancreatitis with SNAP fPL and Spec fPL

Objectives Pancreatitis is a frequent disease in cats for which the ante-mortem diagnosis remains challenging. Feline pancreatic lipase immunoreactivity (fPLI) has been reported to have a high sensitivity for the diagnosis of pancreatitis. The aim of this study was to compare the rapid in-house test SNAP fPL with the standard test Spec fPL and to evaluate the use of SNAP fPL to diagnose pancreatitis in an emergency setting. Methods fPLI of 111 cats with a clinical suspicion of pancreatitis was measured with both SNAP fPL and Spec fPL. Furthermore, clinical signs, haematological and biochemical changes, and abdominal ultrasound findings were recorded. Results Seventy-eight of 111 cats (70.3%) were tested below the cut-off level for pancreatitis with SNAP, as well as Spec fPL, whereas 21/111 (18.9%) were tested with values above the cut-off level with both tests. In 12/111 (10.8%) cats the results were discordant. The comparison of both tests revealed an agreement of 78/80 (97.5%) when Spec fPL was ⩽3.5 μg/l (negative) and 18/20 (90%) when Spec fPL was ⩾5.4 μg/l (positive). The most common clinical signs in cats with suspected pancreatitis (n = 21) were lethargy (95.2%), reduced appetite and vomiting (90.5% each), dehydration (81.0%), diarrhoea (57.1%), abdominal pain and weight loss (47.6% each). Hyperglycaemia and hyperbilirubinaemia (85.7% each), increased aspartate transaminase (76.2%) and alanine transaminase (47.6%), leucocytosis (61.9%), lymphopenia (57.1%), decreased sodium and chloride (57.1% each), and increased urea (52.4%) were the most common abnormalities in blood work. Conclusions and relevance Clinical signs, as well as routine blood-work changes, were non-specific and thus proved to be insufficient to diagnose pancreatitis. The combination of SNAP fPL and subsequent Spec fPL, if indicated, provided the opportunity to rule out or to diagnose pancreatitis with a higher certainty than previously known test methods. This study proved SNAP fPL to be a valuable tool to exclude or include pancreatitis in an emergency settin