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Mucocele formation in dogs is a unique and enigmatic muco-obstructive disease of the gallbladder caused by amassment of abnormal mucus that bears striking pathological similarity to cystic fibrosis. We investigated the role of CFTR in the pathogenesis of this disease. The location and frequency of disease-associated variants in the coding region of CFTR was compared using whole genome sequence data from 2,642 dogs representing breeds at low-risk, high-risk, or with confirmed disease. Expression, localization, and ion transport activity of CFTR was quantified in control and mucocele gallbladders by NanoString, Western blotting, immunofluorescence imaging, and studies in Ussing chambers. Our results establish significant loss of CFTR-dependent anion secretion by mucocele gallbladder mucosa. A significantly lower quantity of CFTR protein was demonstrated relative to E-cadherin in mucocele compared to control gallbladder mucosa. Immunofluorescence identified CFTR along the apical membrane of epithelial cells in control gallbladders but not in mucocele gallbladder epithelium. Decreases in mRNA copy number for CFTR was accompanied by decreases in mRNA for the Cl-/HCO3- exchanger SLC26A3, K+ channels (KCNQ1, KCNN4), and vasoactive intestinal polypeptide receptor (VIPR1) which suggest a driving force for change in secretory function of gallbladder epithelial cells in the pathogenesis of mucocele formation. There were no significant differences in CFTR gene variant frequency, type, or predicted impact comparing low risk, high risk, and definitively diagnosed groups of dogs. This study describes a unique, naturally occurring muco-obstructive disease of the canine gallbladder, with uncanny similarity to cystic fibrosis, and driven by underlying failure of CFTR function.
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Conjoining of the major pancreatic duct and common bile duct at the major duodenal papilla (MDP) is suspected to predispose cats to the clinical syndrome of "triaditis." However, microanatomy of the MDP or presence of lesions at the MDP has not been assessed in cats with or without triaditis. The aims of this study were to characterize feline MDP histomorphology and to identify associations between MDP anatomy/disease and the presence of biliary, pancreatic, or intestinal inflammation or neoplasia. Histologic assessment was prospectively performed on the MDP, duodenum, jejunum, ileum, liver, and pancreas from 124 client-owned cats undergoing postmortem examination. The majority of cats (104/124, 84%) had a complex ductular network at the MDP, with no distinction between pancreatic and common bile ducts. Lymphoid aggregates at the MDP were common (63/124, 51%). Inflammation of the MDP (MDPitis) was present in 35 of 124 cats (28%) and was often concurrent with cholangitis, pancreatitis, or enteritis (32/35, 91%), but was only associated with enteritis (19/35, 54%, P < .05). Triaditis was less common (19/124, 15%), but was associated with both conjoined MDP anatomy (19/19, 100%, P < .05) and MDPitis (12/19, 63%, P < .05). Neoplasia was present in 37 of 124 cats (29%), with lymphoma (28/37, 78%) predominating. Enteropathy-associated T-cell lymphoma type 2 (EATL2) was most common (n = 16/37, 43%) and was associated with triaditis and MDPitis (P < .05). These findings suggest that anatomy, immune activation, and/or inflammation of the MDP may play a role in the pathogenesis of triaditis. Further studies are needed to elucidate the relationships between triaditis, MDPitis, and EATL2.
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Ampola Hepatopancreática , Doenças do Gato , Enterite , Neoplasias , Humanos , Gatos , Animais , Ampola Hepatopancreática/patologia , Pâncreas , Inflamação/patologia , Inflamação/veterinária , Enterite/patologia , Enterite/veterinária , Neoplasias/patologia , Neoplasias/veterinária , Doenças do Gato/patologiaRESUMO
Typical enteropathogenic Escherichia coli (tEPEC) is a leading cause of diarrhea and associated death in children worldwide. Atypical EPEC (aEPEC) lacks the plasmid encoding bundle-forming pili and is considered less virulent, but the molecular mechanism of virulence is poorly understood. We recently identified kittens as a host for aEPEC where intestinal epithelial colonization was associated with diarrheal disease and death. The purposes of this study were to (i) determine the genomic similarity between kitten aEPEC and human aEPEC isolates and (ii) identify genotypic or phenotypic traits associated with virulence in kitten aEPEC. We observed no differences between kitten and human aEPEC in core genome content or gene cluster sequence identities, and no distinguishing genomic content was observed between aEPEC isolates from kittens with nonclinical colonization (NC) versus those with lethal infection (LI). Variation in adherence patterns and ability to aggregate actin in cultured cells mirrored descriptions of human aEPEC. The aEPEC isolated from kittens with LI were significantly more motile than isolates from kittens with NC. Kittens may serve as a reservoir for aEPEC that is indistinguishable from human aEPEC isolates and may provide a needed comparative animal model for the study of aEPEC pathogenesis. Motility seems to be an important factor in pathogenesis of LI associated with aEPEC in kittens.
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Gatos/genética , Escherichia coli Enteropatogênica/genética , Infecções por Escherichia coli/genética , Proteínas de Escherichia coli/genética , Genômica , Sorotipagem , Virulência/genética , Adolescente , Animais , Criança , Pré-Escolar , Infecções por Escherichia coli/microbiologia , Feminino , Variação Genética , Humanos , Lactente , Recém-Nascido , Masculino , SorogrupoRESUMO
Diarrhea is responsible for the death of approximately 900,000 children per year worldwide. In children, typical enteropathogenic Escherichia coli (EPEC) is a common cause of diarrhea and is associated with a higher hazard of death. Typical EPEC infection is rare in animals and poorly reproduced in experimental animal models. In contrast, atypical EPEC (aEPEC) infection is common in both children and animals, but its role in diarrhea is uncertain. Mortality in kittens is often attributed to diarrhea, and we previously identified enteroadherent EPEC in the intestines of deceased kittens. The purpose of this study was to determine the prevalence and type of EPEC in kittens and whether infection was associated with diarrhea, diarrhea-related mortality, gastrointestinal pathology, or other risk factors. Kittens with and without diarrhea were obtained from two shelter facilities and determined to shed atypical EPEC at a culture-based prevalence of 18%. In contrast, quantitative PCR detected the presence of the gene for intimin (eae) in feces from 42% of kittens. aEPEC was isolated from kittens with and without diarrhea. However, kittens with diarrhea harbored significantly larger quantities of aEPEC than kittens without diarrhea. Kittens with aEPEC had a significantly greater severity of small intestinal and colonic lesions and were significantly more likely to have required subcutaneous fluid administration. These findings identify aEPEC to be prevalent in kittens and a significant primary or contributing cause of intestinal inflammation, diarrhea, dehydration, and associated mortality in kittens.
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Doenças do Gato/microbiologia , Diarreia/veterinária , Escherichia coli Enteropatogênica/isolamento & purificação , Infecções por Escherichia coli/mortalidade , Animais , Gatos , Colo/microbiologia , Diarreia/microbiologia , Diarreia/mortalidade , Modelos Animais de Doenças , Escherichia coli Enteropatogênica/genética , Escherichia coli Enteropatogênica/patogenicidade , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Intestino Delgado/microbiologia , Reação em Cadeia da Polimerase Multiplex , Nutrição Parenteral/métodos , Virulência/genéticaRESUMO
BACKGROUND: Gallbladder disease in people is frequently associated with disorders of lipid metabolism and metabolic syndrome. A recently emergent gallbladder disease of dogs, referred to as mucocele formation, is characterized by secretion of abnormal mucus by the gallbladder epithelium and is similarly associated with hyperlipidemia, endocrinopathy, and metabolic dysfunction. The cause of gallbladder mucocele formation in dogs is unknown. METHODS: A prospective case-controlled study was conducted to gain insight into disease pathogenesis by characterization of plasma lipid abnormalities in 18 dogs with gallbladder mucocele formation and 18 age and breed matched control dogs using direct infusion mass spectrometry for complex plasma lipid analysis. This analysis was complemented by histochemical and ultrastructural examination of gallbladder mucosa from dogs with gallbladder mucocele formation and control dogs for evidence of altered lipid homeostasis of the gallbladder epithelium. RESULTS: Gallbladder mucocele formation in dogs carried a unique lipidomic signature of increased lipogenesis impacting 50% of lipid classes, 36% of esterified fatty acid species, and 11% of complex lipid species. Broad enrichment of complex lipids with palmitoleic acid (16:1) and decreased abundance within complex lipids of presumptive omega-3 fatty acids eicosapentaenoic (20:5) and docosahexaenoic (22:6) was significant. Severe lipidosis of gallbladder epithelium pinpoints the gallbladder as involved causally or consequently in abnormal lipid metabolism. CONCLUSION: Our study supports a primary increase in lipogenesis in dogs with mucocele formation and abnormal gallbladder lipid metabolism in disease pathogenesis.
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Doenças do Cão , Doenças da Vesícula Biliar , Vesícula Biliar , Lipogênese , Mucocele , Animais , Cães , Mucocele/metabolismo , Mucocele/patologia , Vesícula Biliar/metabolismo , Vesícula Biliar/patologia , Doenças do Cão/metabolismo , Doenças do Cão/patologia , Doenças da Vesícula Biliar/metabolismo , Doenças da Vesícula Biliar/patologia , Doenças da Vesícula Biliar/veterinária , Feminino , Estudos de Casos e Controles , Masculino , Lipidoses/metabolismo , Lipidoses/patologia , Estudos Prospectivos , Epitélio/metabolismo , Epitélio/patologia , Metabolismo dos LipídeosRESUMO
Protozoal diarrhea caused by Tritrichomonas foetus (blagburni) is a prevalent, lifelong, and globally distributed burden in domestic cats. Treatment is limited to the use of 5-nitroimidazoles and treatment failure is common. The repurposed gold salt compound auranofin has killing activity against diverse protozoa in vitro but evidence of efficacy in naturally occurring protozoal infections is lacking. This exploratory study investigated the efficacy and safety of auranofin for treatment of cats with naturally occurring, 5-nitroimidazole-resistant, T. foetus infection. The minimum lethal concentration (MLC) of auranofin against 5 isolates of feline T. foetus was determined under aerobic conditions in vitro. Healthy cats and cats with T. foetus infection were treated with immediate release auranofin (range, 0.5-3â¯mg/cat for 7 days) or guar gum-coated auranofin capsules (0.5 or 3â¯mg/cat for 7 days). Adverse effects were monitored by clinical signs and clinicopathologic testing. Efficacy was determined by fecal consistency score, bowel movement frequency, and single-tube nested PCR of feces for T. foetus rDNA. Fecal samples were assayed for concentrations of auranofin, known and predicted metabolites of auranofin, gold containing molecules, and total gold content using HPLC, LC-MS, ion mobility-MS, and ICP-MS, respectively. Auranofin was effective at killing isolates of feline T. foetus at MLC ≥ 1 µg/ml. Treatment of cats with T. foetus infection with either immediate release auranofin or a colon-targeted guar gum-coated tablet of auranofin did not eradicate infection. Treatment failure occurred despite fecal concentrations of gold that met or exceeded the equivalent MLC of auranofin. Neither auranofin, known or predicted metabolites of auranofin, nor any gold-containing molecules >100â¯Da could be detected in fecal samples of treated cats. Adverse effects associated with auranofin treatment were common but minor. These studies identify that in vitro susceptibility test results of auranofin may not translate to treatment effectiveness in vivo even when achieving gold concentrations equivalent to the MLC of auranofin in the target environment. These studies further establish the absence of any predicted or unpredicted gold containing metabolites in feces after oral administration of auranofin.
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BACKGROUND & AIMS: Although we are beginning to understand where, when, and how intestinal epithelial cells are shed, physiologically, less is understood about alterations in cell fate during minimally invasive epithelial infections. We used a piglet model of Cryptosporidium parvum infection to determine how elimination of infected enterocytes is balanced with the need to maintain barrier function. METHODS: We studied the effects of enterocyte shedding by C parvum-infected ileum on barrier function ex vivo with Ussing chambers. The locations and activities of caspase-3, nuclear factor κB (NF-κB), and inhibitor of apoptosis proteins (IAP) were assayed by enzyme-linked immunosorbent assay, immunoblot, and tissue immunoreactivity analyses and using specific pharmacologic inhibitors. The location, specificity, and magnitude of enterocyte shedding were quantified using special stains and light microscopy. RESULTS: Infection with C parvum activated apoptotic signaling pathways in enterocytes that resulted in cleavage of caspase-3. Despite caspase-3 cleavage, enterocyte shedding was confined to villus tips, coincident with apoptosis, and observed more frequently in infected cells. Epithelial expression of X-linked inhibitor of apoptosis protein (XIAP), activation of NF-κB, and proteasome activity were required for control of cell shedding and barrier function. The proteasome blocked activity of caspase-3; this process was mediated by expression of XIAP, which bound to cleaved caspase-3. CONCLUSIONS: We have identified a pathway by which villus epithelial cells are maintained during C parvum infection. Loss of barrier function is reduced by active retention of infected enterocytes until they reach the villus tip. These findings might be used to promote clearance of minimally invasive enteropathogens, such as by increasing the rate of migration of epithelial cells from the crypt to the villus tip.
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Enterócitos/metabolismo , Íleo/fisiopatologia , Mucosa Intestinal/metabolismo , NF-kappa B/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismo , Animais , Apoptose , Caspase 3/metabolismo , Criptosporidiose/metabolismo , Cryptosporidium parvum , Modelos Animais de Doenças , Enterócitos/microbiologia , Íleo/microbiologia , Transdução de Sinais , SuínosRESUMO
Approximately 15% of foster kittens die before 8 weeks of age, with most of these kittens demonstrating clinical signs or postmortem evidence of enteritis. While a specific cause of enteritis is not determined in most cases, these kittens are often empirically administered probiotics that contain enterococci. The enterococci are members of the commensal intestinal microbiota but also can function as opportunistic pathogens. Given the complicated role of enterococci in health and disease, it would be valuable to better understand what constitutes a "healthy" enterococcal community in these kittens and how this microbiota is impacted by severe illness. In this study, we characterized the ileum mucosa-associated enterococcal community of 50 apparently healthy and 50 terminally ill foster kittens. In healthy kittens, Enterococcus hirae was the most common species of ileum mucosa-associated enterococci and was often observed to adhere extensively to the small intestinal epithelium. These E. hirae isolates generally lacked virulence traits. In contrast, non-E. hirae enterococci, notably Enterococcus faecalis, were more commonly isolated from the ileum mucosa of kittens with terminal illness. Isolates of E. faecalis had numerous virulence traits and multiple antimicrobial resistances. Moreover, the attachment of Escherichia coli to the intestinal epithelium was significantly associated with terminal illness and was not observed in any kitten with adherent E. hirae. These findings identify a significant difference in the species of enterococci cultured from the ileum mucosa of kittens with terminal illness compared to the species cultured from healthy kittens. In contrast to prior case studies that associated enteroadherent E. hirae with diarrhea in young animals, these controlled studies identified E. hirae as more often isolated from healthy kittens and adherence of E. hirae as more common and extensive in healthy kittens than in sick kittens.
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Biota , Doenças do Gato/microbiologia , Enterite/veterinária , Enterococcus/isolamento & purificação , Escherichia coli/isolamento & purificação , Íleo/microbiologia , Mucosa Intestinal/microbiologia , Animais , Animais Recém-Nascidos , Biofilmes/crescimento & desenvolvimento , Doenças do Gato/mortalidade , Gatos , Enterite/microbiologia , Enterite/mortalidade , Enterococcus/fisiologia , Escherichia coli/fisiologiaRESUMO
BACKGROUND: Bacterial infection of bile is a common cause of hepatobiliary disease in cats. Whether bile harbors a core microbiota in health or in cases of suspected hepatobiliary disease in cats is unknown. OBJECTIVES: Establish if gallbladder bile in apparently healthy cats harbors a core microbiota composed of bacterial taxa common to many individuals. Compare results of bile cytology, bile culture, and 16S rRNA gene amplicon sequencing in apparently healthy cats and cats with suspected hepatobiliary disease. ANIMALS: Forty-three client-owned cats with suspected hepatobiliary disease and 17 control cats. METHODS: Bile was collected by ultrasound guided cholecystocentesis (cats with suspected hepatobiliary disease) or laparotomy after euthanasia (controls). Bile samples underwent cytologic examination, aerobic and anaerobic culture, and DNA was extracted for 16S rRNA gene amplification and sequencing. RESULTS: Microbiome sequencing did not identify a core microbiota in control cats or cats having bile sampled because of clinical suspicion for hepatobiliary disease. Microbiome profiles from control cats were indistinguishable from profiles obtained from sampling instruments and reagents that were not exposed to bile (technical controls). Bacterial taxa that could not be explained by contamination or off-target amplification were identified only in samples from cats with bactibilia and positive bile culture results for Escherichia coli. In several E. coli positive samples, microbiome sequencing also identified a small number of potentially co-infecting bacterial genera not identified by culture. CONCLUSIONS AND CLINICAL IMPORTANCE: Cat bile does not harbor a core microbiota. Uncultured bacteria may contribute to pathogenesis of hepatobiliary disease in cats with bile E. coli infection.
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Doenças do Gato , Doenças do Sistema Digestório , Infecções por Escherichia coli , Microbiota , Humanos , Gatos , Animais , Bile , Escherichia coli , RNA Ribossômico 16S/genética , Doenças do Sistema Digestório/veterinária , Infecções por Escherichia coli/veterináriaRESUMO
To date studies have not investigated the culture-independent microbiome of bile from dogs, a species where aseptic collection of bile under ultrasound guidance is somewhat routine. Despite frequent collection of bile for culture-based diagnosis of bacterial cholecystitis, it is unknown whether bile from healthy dogs harbors uncultivable bacteria or a core microbiota. The answer to this question is critical to understanding the pathogenesis of biliary infection and as a baseline to exploration of other biliary diseases in dogs where uncultivable bacteria could play a pathogenic role. A pressing example of such a disease would be gallbladder mucocele formation in dogs. This prevalent and deadly condition is characterized by excessive secretion of abnormal mucus by the gallbladder epithelium that can eventually lead to rupture of the gallbladder or obstruction of bile flow. The cause of mucocele formation is unknown as is whether uncultivable, and therefore unrecognized, bacteria play any systematic role in pathogenesis. In this study we applied next-generation 16S rRNA gene sequencing to identify the culture-negative bacterial community of gallbladder bile from healthy dogs and gallbladder mucus from dogs with mucocele formation. Integral to our study was the use of 2 separate DNA isolations on each sample using different extraction methods and sequencing of negative control samples enabling recognition and curation of contaminating sequences. Microbiota findings were validated by simultaneous culture-based identification, cytological examination of bile, and fluorescence in-situ hybridization (FISH) performed on gallbladder mucosa. Using culture-dependent, cytological, FISH, and 16S rRNA sequencing approaches, results of our study do not support existence of a core microbiome in the bile of healthy dogs or gallbladder mucus from dogs with mucocele formation. Our findings further document how contaminating sequences can significantly contribute to the results of sequencing analysis when performed on samples with low bacterial biomass.
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Doenças dos Ductos Biliares , Doenças do Cão , Doenças da Vesícula Biliar , Microbiota , Mucocele , Cães , Animais , Vesícula Biliar/patologia , Mucocele/veterinária , RNA Ribossômico 16S/genética , Bile/microbiologia , Doenças da Vesícula Biliar/veterinária , Microbiota/genética , Doenças do Cão/diagnósticoRESUMO
Introduction: Diarrhea is the second most common cause of mortality in shelter kittens. Studies examining prevention strategies in this population are lacking. Probiotics are of particular interest but studies in cats are largely limited to healthy adults or those with induced disease. Only one study in domestic cats describes the use of host-derived bacteria as a probiotic. We previously identified Enterococcus hirae as a dominant species colonizing the small intestinal mucosa in healthy shelter kittens. Oral administration of a probiotic formulation of kitten-origin E. hirae (strain 1002-2) mitigated the increase in intestinal permeability and fecal water loss resulting from experimental enteropathogenic E. coli infection in purpose-bred kittens. Based on these findings, we hypothesized that administration of kitten-origin E. hirae to weaned fostered shelter kittens could provide a measurable preventative health benefit. Methods: We conducted a randomized, placebo-controlled, blinded clinical trial to determine the impact of a freeze-dried E. hirae probiotic on body weight gain, incidence of diarrhea, carriage of potential diarrheal pathogens, and composition of the intestinal microbiota in weaned fostered shelter kittens. Results: One-hundred thirty kittens completed the study. Fifty-eight kittens received the probiotic and 72 received the placebo. There were no significant differences in age, weight upon initiation of the study, number of days in the study, average daily gain in body weight, or weight at completion of the study. Kittens treated with E. hirae were 3.4 times less likely to develop diarrhea compared to kittens treated with placebo (odds ratio = 0.294, 95% CI 0.109-0.792, p = 0.022). A significant impact of E. hirae was not observed on the presence or abundance of 30 different bacterial, viral, protozoal, fungal, algal, and parasitic agents in feces examined by qPCR. With exception to a decrease in Megamonas, administration of the E. hirae probiotic did not alter the predominant bacterial phyla present in feces based on 16S rRNA gene amplicon sequencing. Discussion: Decreased incidence of diarrhea associated with preventative administration of E. hirae to foster kittens supports a rationale for use of E. hirae for disease prevention in this young population at high risk for intestinal disease though additional studies are warranted.
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During early urinary tract infection (UTI) the interplay between invading bacteria and the urothelium elicits a mucosal response aimed at clearing infection. Unfortunately, the resultant inflammation and associated local tissue injury are responsible for patient symptoms. Interleukin-6 (IL-6), a cytokine released during acute UTI, has both pro- and anti-inflammatory effects on other body systems. Within the urothelium, the IL-6 native-tissue origin, the target cell type(s), and ultimate effect of the cytokine on target cells are largely unknown. In the present study we modeled the UTI IL-6 response ex vivo using canine bladder mucosa mounted in Ussing chambers to determine the inflammatory and reparative role of IL-6. We demonstrated that uropathogenic Escherichia coli infection stimulates the synthesis of IL-6 by all urothelial cell layers, with the urothelial cells alone representing the only site of unequivocal IL-6 receptor expression. Autocrine effects of IL-6 were supported by the activation of urothelial STAT3 signaling and SOCS3 expression. Using exogenous IL-6, a microarray approach, and quantitative reverse transcriptase PCR (q-RT-PCR), 5 target genes (tumor necrosis factor alpha, interleukin-1ß, matrix metallopeptidase 2, heparan sulfate d-glucosaminyl 3-O-sulfotransferase 3A1, and hyaluronan synthase 2) that have direct or indirect roles in promoting a proinflammatory state were identified. Two of these genes, heparan sulfate d-glucosaminyl 3-O-sulfotransferase 3A1 and hyaluronan synthase 2, are also potentially important mediators of wound repair via the production of glycosaminoglycan components. These findings suggest that IL-6 secretion during acute UTI may serve a dual biological role by initiating the inflammatory response while also repairing urothelial defenses.
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Reação de Fase Aguda/metabolismo , Interleucina-6/metabolismo , Bexiga Urinária/metabolismo , Animais , Cães , Humanos , Mucosa/metabolismo , Mucosa/microbiologia , Transdução de Sinais , Técnicas de Cultura de Tecidos , Bexiga Urinária/microbiologia , Escherichia coli Uropatogênica , Urotélio/metabolismoRESUMO
The protozoal pathogen Tritrichomonas foetus infects the colon of domestic cats and is a major cause of chronic colitis and diarrhea. Treatment failure is common, but antibiotics may improve clinical signs in a subset of cats, leading researchers to question involvement of the colonic microbiota in disease pathogenesis. Studies performed in women with venereal Trichomonas vaginalis infections have revealed that dysbiosis of host microbiota contributes to pathogenicity with similar findings also found in mice with intestinal Tritrichomonas musculis The aim of this study was to characterize differences in the fecal microbiota of cats with and without naturally occurring T. foetus infection and in a group of kittens prior to and after experimentally induced infection. Archived fecal DNA from cats undergoing testing for T. foetus infection (n = 89) and experimentally infected kittens (n = 4; at pre-, 2 weeks, and 9 weeks post-infection) were analyzed by sequencing of 16S rRNA genes. Amongst the naturally infected population, the genera Megamonas and Helicobacter were significantly increased in prevalence and abundance in cats testing positive for T. foetus infection. In the group of four experimentally infected kittens, fecal samples post-infection had significantly lower abundance of genus Dialister and Megamonas and greater abundance of the class Betaproteobacteria and family Succinivibrionaceae. We hypothesize that T. foetus promotes dysbiosis by competition for fermentable substrates used by these bacteria and that metabolic byproducts may contribute to the pathogenesis of colonic inflammation and diarrhea. Future studies are warranted for the measurement of fecal concentrations of microbial and protozoal metabolites in cats with T. foetus infection for the identification of potential therapeutic targets.
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Disbiose/complicações , Fezes/microbiologia , Microbiota , Infecções por Protozoários/complicações , Tritrichomonas foetus/fisiologia , Animais , Gatos , Modelos Animais de Doenças , Camundongos , Infecções por Protozoários/microbiologiaRESUMO
BACKGROUND: Proteinuria is an independent risk factor for morbidity and mortality in dogs. An association between proteinuria and gallbladder mucocele formation in dogs is unknown. OBJECTIVE: Determine if gallbladder mucocele formation or clinicopathologic comorbidities are associated with proteinuria. ANIMALS: Twenty-five dogs with mucocele formation and 25 breed and age-matched control dogs from a prior study. METHODS: Retrospective case control study. Proteinuria defined by calculated urine dipstick protein concentration (mg/mL) to urine specific gravity (USG) ratio. Clinicopathologic findings, postcosyntropin cortisol concentration, thyroid function profile, and illness severity score were recorded. RESULTS: Median urine dipstick protein concentration to USG ratio and number of dogs having a ratio ≥1.5 were significantly higher for dogs with mucocele formation compared to control dogs. Proteinuria was not significantly associated with CBC or serum biochemistry profile abnormalities but increased in relation to severity of illness. CONCLUSIONS AND CLINICAL IMPORTANCE: Gallbladder mucocele formation is significantly associated with proteinuria in dogs. Diagnosis and treatment of proteinuria in dogs with mucocele formation might minimize long term kidney morbidity in these patients.
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Doenças do Cão , Mucocele , Animais , Estudos de Casos e Controles , Doenças do Cão/etiologia , Cães , Vesícula Biliar , Mucocele/complicações , Mucocele/veterinária , Proteinúria/veterinária , Estudos RetrospectivosRESUMO
The bacterial causes of diarrhea can be frustrating to identify, and it is likely that many remain undiagnosed. The pathogenic potential of certain bacteria becomes less ambiguous when they are observed to intimately associate with intestinal epithelial cells. In the present study we sought to retrospectively characterize the clinical, in situ molecular, and histopathological features of enteroadherent bacteria in seven unrelated kittens that were presumptively diagnosed with enteropathogenic Escherichia coli (EPEC) on the basis of postmortem light microscopic and, in some cases, microbiological examination. Characterization of the enteroadherent bacteria in each case was performed by Gram staining, in situ hybridization using fluorescence-labeled oligonucleotide probes, PCR amplification of species-specific gene sequences, and ultrastructural imaging applied to formalin-fixed paraffin-embedded sections of intestinal tissue. In only two kittens was EPEC infection confirmed. In the remaining five kittens, enteroadherent bacteria were identified as Enterococcus spp. The enterococci were further identified as Enterococcus hirae on the basis of PCR amplification of DNA extracted from the formalin-fixed, paraffin-embedded tissue and amplified by using species-specific primers. Transmission electron microscopy of representative lesions from E. coli- and Enterococcus spp.-infected kittens revealed coccobacilli adherent to intestinal epithelial cells without effacement of microvilli or cup-and-pedestal formation. Enterococci were not observed, nor were DNA sequences amplified from intestinal tissue obtained from age-matched kittens euthanized for reasons unrelated to intestinal disease. These studies suggest that E. hirae may be a common cause of enteroadherent bacterial infection in pre-weaning-age kittens and should be considered in the differential diagnosis of bacterial disease in this population.
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Doenças do Gato/diagnóstico , Doenças do Gato/microbiologia , Enterococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/veterinária , Hibridização in Situ Fluorescente/métodos , Enteropatias/veterinária , Animais , Técnicas Bacteriológicas/métodos , Doenças do Gato/patologia , Gatos , Enterococcus/classificação , Enterococcus/genética , Infecções por Bactérias Gram-Positivas/diagnóstico , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/patologia , Histocitoquímica , Enteropatias/diagnóstico , Enteropatias/microbiologia , Enteropatias/patologia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Microscopia , Microscopia Eletrônica de Transmissão , Reação em Cadeia da Polimerase , Estudos RetrospectivosRESUMO
We hypothesized that unrecognized differences in epithelial expression of inducible nitric oxide synthase (iNOS), resulting from engineered immunodeficiency, could explain the contradictory findings of prior studies regarding the importance of nitric oxide (NO) in murine models of Cryptosporidium parvum infection. Severe combined immunodeficient mice (SCID) failed to constitutively or inducibly express epithelial iNOS or increase NO synthesis in response to C. parvum infection. In contrast, mice lacking IFN-gamma alone induced both epithelial iNOS expression and NO synthesis in response to infection. Accordingly, lymphocytes mediate epithelial expression of iNOS and NO synthesis independent of IFN-gamma in response to C. parvum infection. These findings in large part explain the contradictory conclusions of prior studies regarding the role of iNOS in C. parvum infection.
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Criptosporidiose/imunologia , Cryptosporidium parvum/imunologia , Interferon gama/imunologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Linfócitos/imunologia , Óxido Nítrico Sintase Tipo II/biossíntese , Animais , Feminino , Interferon gama/deficiência , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Camundongos SCIDRESUMO
The regulation of the glomerular filtration rate (GFR) is a particularly important and challenging concept for students to integrate into a memorable framework for building further knowledge and solving clinical problems. In this study, 76 first-year veterinary students and 19 veterinarians in clinical specialty training (house officers) participated in separate online exercises to evaluate the use of a computer-animated model of GFR regulation (www.aamc.org/mededportal) on learning outcome. Students were randomly allocated to study either the animated model or written materials before completion of a 10-question multiple-choice quiz. House officers completed a 35-question test before and after study of the animated model. Both groups completed a survey about the learning exercise. The ability of the model to enhance learning was demonstrated by a significant improvement (P < 0.001) in the test performance of house officers after studying the model. The model performed similarly to written materials alone in affecting the subsequent quiz performance of the students. The majority of students and house officers agreed or strongly agreed that the animated model was easy to understand, improved their knowledge and appreciation of the importance of GFR regulation, and that they would recommend the model to peers. Most students [63 of 76 students (83%)] responded that they would prefer the use of the animated model alone over the study of written materials but acknowledged that a combination of hardcopy written notes and the animated model would be ideal. A greater applicability of the model to more advanced students and an introduction in a didactic setting before individual study were suggested by the house officers. The results of this study suggest that the animated model is a useful, effective, and well-received tool for learning and creating a visual memory of the regulatory mechanisms of GFR.
Assuntos
Simulação por Computador , Educação em Veterinária/métodos , Taxa de Filtração Glomerular/fisiologia , Fisiologia/educação , Aprendizagem Baseada em Problemas/métodos , HumanosRESUMO
OBJECTIVE: To determine whether Tritrichomonas foetus infection resides in reproductive tract tissues from cats housed for breeding and for which a high prevalence of colonic T foetus infection has been reported. ANIMALS: 61 purebred cats in 36 catteries undergoing elective ovariohysterectomy or castration and for which reproductive tract tissues, feces, and a reproductive history were obtained. PROCEDURES: Reproductive tract tissues were examined for T foetus via light microscopy, immunohistochemical analysis, and PCR assay. History of reproductive tract disease was examined to detect statistical associations with identified or reported exposure to colonic T foetus infection. RESULTS: 15 of 61 (25%) cats and 22 of 33 (67%) catteries were identified with active or reported T foetus infection. Light microscopic, immunohistochemical, or molecular evidence of T foetus infection of the reproductive tract was not detected in any cats, including 15 cats with colonic T foetus infection, 29 cats residing in a cattery in which T foetus-infected cats were identified, and 8 cats for which gross or light microscopic evidence of reproductive tract disease was identified. There were no differences in total number of litters, number of litters per breeding, kitten mortality rate, or birth defects between cats or catteries infected with T foetus and those for which T foetus infection was not identified. CONCLUSIONS AND CLINICAL RELEVANCE: No evidence of reproductive tract colonization by T foetus was detected in this study. Accordingly, it is unlikely that reproductive tract infection with T foetus plays an important role in overall disease transmission.
Assuntos
Doenças do Gato/parasitologia , Doenças dos Genitais Femininos/veterinária , Doenças dos Genitais Masculinos/veterinária , Infecções Protozoárias em Animais/parasitologia , Tritrichomonas foetus , Animais , Antiprotozoários/uso terapêutico , Gatos , Feminino , Doenças dos Genitais Femininos/parasitologia , Doenças dos Genitais Masculinos/parasitologia , Masculino , Ronidazole/uso terapêuticoRESUMO
BACKGROUND: Fecal polymerase chain reaction (PCR) testing for Tritrichomonas foetus is considered the most sensitive means for diagnosis of infection but results could be influenced by fecal collection technique and prior use of antimicrobial drugs. OBJECTIVES: To establish any association between fecal collection technique or treatment history and results of fecal PCR testing for T. foetus. ANIMALS: Fecal samples from 1717 cats submitted by veterinarians between January 2012 and December 2017. METHODS: This study used a retrospective analysis. T. foetus PCR test results from 1808 fecal samples submitted for diagnostic testing were examined for their association with method of fecal collection and prior antimicrobial treatments. Data were collected from sample submission form. RESULTS: Positive T. foetus PCR test results were obtained for 274 (16%) cats. Fecal samples collected via fecal loop had increased probability of positive PCR test results (odds ratio [OR] 2.04, 95% confidence interval [CI] 1.31-3.17, P = .002) compared to samples collected by colonic flush. There was no association between PCR test results and treatment history, treatment type, or prior treatment with ronidazole. After an initial positive PCR test, 4/19 (21%; 95% CI 2.7%-39.4%) cats treated with ronidazole had a second positive test result. CONCLUSIONS AND CLINICAL IMPORTANCE: Results of this study support that fecal samples collected by loop might be better for PCR diagnosis of T. foetus infection. Lack of association of ronidazole with PCR test results and a 21% all-potential-causes failure rate of ronidazole in cats with preconfirmed infection are important limitations to use of this drug.
Assuntos
Doenças do Gato/diagnóstico , Fezes/parasitologia , Infecções Protozoárias em Animais/diagnóstico , Manejo de Espécimes/veterinária , Tritrichomonas foetus/isolamento & purificação , Animais , Antiprotozoários/uso terapêutico , Doenças do Gato/tratamento farmacológico , Doenças do Gato/parasitologia , Gatos , Feminino , Masculino , Reação em Cadeia da Polimerase/veterinária , Infecções Protozoárias em Animais/tratamento farmacológico , Infecções Protozoárias em Animais/parasitologia , Estudos Retrospectivos , Ronidazole/uso terapêutico , Estados UnidosRESUMO
OBJECTIVES: The objective of this study was to identify significant associations between treatment interventions and survival of orphaned shelter kittens with diarrhea. METHODS: Orphaned kittens admitted to a large open-intake municipal animal shelter and entering a volunteer foster care provider network between April 2016 and July 2017 were studied. Individual orphaned kittens for which the care provider sought examination by shelter veterinarians because of clinical signs of diarrhea were included. Treatments administered were recorded and their association with kitten survival to adoption or transfer to a rescue partner was statistically examined. RESULTS: A total of 1718 orphaned kittens were fostered, among which 220 kittens (12.8%) from 118 different litters were presented for evaluation of diarrhea. A total of 172 (78.2%) kittens underwent treatment for their diarrhea, among which 153 (89.0%) survived to adoption or transfer to a rescue partner and 19 (11.0%) died or were humanely euthanized. Kittens with diarrhea that were ⩾4 weeks of age were 24.8 times more likely to survive (95% confidence interval [CI] 6.8-89.8; P <0.0001). While controlling for age, kittens that received a vitamin and mineral supplement were 12.8 times more likely to survive (95% CI 3.1-52.5; P = 0.0004) than kittens with diarrhea that did not receive the supplement. Treatment with subcutaneous fluids, penicillin G, tube feeding, a probiotic containing Enterococcus faecium SF68, ponazuril or metronidazole did not statistically significantly increase the survival of kittens with diarrhea. CONCLUSIONS AND RELEVANCE: Age ⩾4 weeks and treatment of diarrhea with a vitamin and mineral supplement favorably improves survival of orphaned kittens to adoption. Application of these findings are likely to improve the health and welfare of this population and contribute to a refinement in use of shelter resources.