Hair Sample Analysis as a Method of Monitoring Exposure to Bisphenol A in Dogs.

Bisphenol A (BPA) is an organic substance widely used in the plastics industry. It penetrates food and environment and, as an endocrine disruptor, has detrimental effects on human organisms. Pet animals, which live in the immediate vicinity of humans, are also exposed to BPA; however, knowledge regarding the exposure of dogs to this substance is extremely scarce. This is the first study in which hair analysis has been used to biomonitor BPA in 30 dogs using liquid chromatography and tandem mass spectrometry techniques. The presence of BPA in concentration levels above the method detection limit (1.25 ng/g) was noted in 93.33% of samples. BPA concentration levels were found to range from 7.05 ng/g to 436 ng/g (mean 81.30 ng/g). Statistically significant differences in BPA concentration levels were found between animals with physiological weight and animals with abnormal weight (skinny and obese). In turn, differences between males and females, as well as between young, middle-aged and old dogs, were not statistically significant. The obtained results have clearly shown that hair analysis is a useful method to evaluate the exposure of dogs to BPA. This study also confirmed that dogs are exposed to BPA to a large extent, and this substance may play a role as a pathological factor in this animal species. However, many aspects connected to the influence of BPA on canine health status are unclear and need further study.

The use of Savary-Gilliard dilators in the treatment of an oesophageal stricture in a cat.

Oesophageal strictures in cats and dogs are relatively rare and the cause of this disorder can be multifactorial. However, the most common cause in cats is an inflammatory process.Conservative treatment strategies for this disorder includes image-guided interventions. Endoscopic methods are a form of a minimally invasive surgical treatment of the oesophageal strictures. Several endoscopic methods for the therapy of this condition are known, one of them is Savary-Gilliard dilators technique.In the present study of a case of oesophageal stricture in a cat, caused probably by doxycycline treatment without water administration, the authors used the Savary-Gilliard dilators as a therapy for its condition. The animal underwent 3 endoscopy procedures, where in the third one no abnormality in the oesophagus was observed. Moreover, the cat was asymptomatic 6 months after the last oesophagoscopy.In the authors opinion, based on the present case, some experience of the authors and previously described studies, the Savary-Gilliard dilators seems to be a safe, effective, relatively cheap and minimally invasive method of the oesophageal stricture therapy in the cat.

Biomonitoring parabens in dogs using fur sample analysis - Preliminary studies.

Parabens are widely used in the food, cosmetics and pharmaceutical industry and are widespread in the environment. As endocrine disruptors, parabens have adverse effects on living organisms. However, knowledge of the exposure of domestic animals to parabens is extremely scarce. Therefore, this study assessed the exposure level of dogs to three parabens commonly used in industry (i.e. methylparaben - MeP, ethylparaben - EtP and propylparaben - PrP) using fur sample analysis in liquid chromatography-tandem mass spectrometry. The presence of parabens has been noted in the samples collected from all dogs included in the study (n = 30). Mean concentrations of MeP, EtP and PrP in the fur of dogs were 176 (relative standard deviation - RSD = 127.48%) ng/g dry weight (dw), 48.4 (RSD = 163.64%) ng/g dw and 79.8 ng/g dw (RSD = 151.89%), respectively. The highest concentrations were found for MeP (up to 1023 ng/g dw). Concentrations of MeP and EtP in males were statistically higher than those in females (p < 0.05). Statistically significantly higher concentration levels of PrP in young animals (up to three years old) were also found. This is the first study concerning the use of fur samples to evaluate the exposure of domestic animals to parabens. The results indicate that an analysis of the fur may be a useful tool of paraben biomonitoring in dogs. The presence of parabens in the canine fur also suggests that these substances may play a role in veterinary toxicology. However, many aspects connected with this issue are not clear and require further study.

Bisphenol A affects vipergic nervous structures in the porcine urinary bladder trigone.

Bisphenol A (BPA) is used in the production of plastics approved for contact with feed and food. Upon entering living organisms, BPA, as a potent endocrine disruptor, negatively affects various internal organs and regulatory systems, especially in young individuals. Although previous studies have described the neurotoxic effects of BPA on various tissues, it should be underlined that the putative influence of this substance on the chemical architecture of the urinary bladder intrinsic innervation has not yet been studied. One of the most important neuronal substances involved in the regulation of urinary bladder functions is vasoactive intestinal polypeptide (VIP), which primarily participates in the regulation of muscular activity and blood flow. Therefore, this study aimed to determine the influence of various doses of BPA on the distribution pattern of VIP-positive neural structures located in the wall of the porcine urinary bladder trigone using the double-immunofluorescence method. The obtained results show that BPA influence leads to an increase in the number of both neurons and nerve fibres containing VIP in the porcine urinary bladder trigone. This may indicate that VIP participates in adaptive processes of the urinary bladder evoked by BPA.

Assessment of exposure to perfluoroalkyl substances (PFASs) in dogs by fur analysis.

Poly- and perfluoroalkyl substances (PFASs) are a large group of chemicals commonly used in various branches of industry, which may adversely affect the living organisms. The aim of this study were to evaluate exposure of dogs to six selected PFASs: five perfluoroalkyl carboxylic acids (perfluorobutanoic acid - PFBuA, perfluoropentanoic acid - PFPeA, perfluorohexanoic acid - PFHxA, perfluoroheptanoic acid - PFHpA, perfluorooctanoic acid - PFOA) and perfluorooctane sulfonic acid (PFOS) through the analysis of fur samples. To our knowledge this is the first study concerning the use of fur samples to evaluation of exposure of domestic animals to PFASs. Relationship between PFASs concentration and age, gender and body weight of animals was also evaluated. Fur samples were collected from 30 dogs living in Olsztyn (Poland). All PFASs studied were detected in the canine fur samples. The highest concentrations were observed in the case of PFOA and PFBuA, detected at concentrations in the range between 1.51 and 66.7 ng/g and 0.98-26.6 ng/g, respectively. During the present study generally no statistically significant differences dependent on gender, age and body weight of animals were found. This study confirms the suitability of fur samples for biomonitoring of exposure to PFASs in domestic animals, what may be important in veterinary toxicology.

Endoscopic Biopsies and Histopathological Findings in Diagnosing Chronic Gastrointestinal Disorders in Dogs and Cats.

Nowadays, endoscopic examination is a diagnostic tool gaining popularity in the management of gastrointestinal disorders in dogs and cats. Direct accessibility of the lumen of gastrointestinal tract combined with the mucosal biopsy provides a great diagnostic potential. Using endoscopy and endoscopically guided biopsy, one can conduct both macro- and microscopic assessment of lesions and perform many specialist adjunct examinations. Histopathological examination of mucosal biopsy specimens collected from the stomach and intestines allows us to distinguish between types of inflammations and to diagnose ulcerative, polypoid, and cancerous lesions.

A Few TH-Immunoreactive Neurons Closely Appose DMX-Located Neuronal Somata Projecting to the Stomach Prepyloric Region in the Pig.

The vagus nerve is responsible for efferent innervation and functional control of stomach functions. The efferent fibers originate from neurons located in the dorsal motor nucleus of the vagus (DMX) and undergo functional control of the local neuroregulatory terminals. The aim of the present study was to examine the existence of morphological foundations for direct regulatory action of the local TH-immunoreactive neurons on parasympathetic efferent neurons supplying the prepyloric region of the porcine stomach. Combined injection of neuronal retrograde tracer Fast Blue into the stomach prepyloric region with TH immunostaining was used in order to visualize spatial relationship between DMX-located stomach prepyloric region supplying neuronal stomata and local TH-IR terminals. We confirmed existence of TH-immunoreactive neural terminals closely opposing the stomach prepyloric region innervating neurons at the porcine DMX area. The observed spatial relationship points out the possibility of indirect catecholaminergic control of the stomach function exerted through preganglionic parasympathetic efferent neurons in the pig.

Vasoactive Intestinal Polypeptide (VIP) in the Intestinal Mucosal Nerve Fibers in Dogs with Inflammatory Bowel Disease.

Canine inflammatory bowel disease (IBD) is a group of enteropathies with nonspecific chronic symptoms and poorly understood etiology. Many aspects connected with IBD are not understood. One of them is the participation of the intestinal nervous system in the development of pathological processes. Thus, this study aimed to demonstrate changes in the density of intramucosal nerve fibers containing vasoactive intestinal polypeptide (VIP)-one of the most important intestinal nervous factors caused by the various stages of IBD development. Mucosal biopsy specimens collected from the duodenum, jejunum and descending colon of healthy dogs and dogs with varied severity of IBD were included in the experiment. The density of VIP-like immunoreactive (VIP-LI) nerves was determined by a single immunofluorescence technique and a semi-quantitative method consisting in VIP-LI fiber counts in the field of view (0.1 mm2). The obtained results indicate that IBD induces changes in the density of mucosal VIP-LI nerve fibers in the canine gastrointestinal tract. The initial decrease is followed by an increase in VIP-like immunoreactivity in successive stages of the disease. These observations show that VIP is a neuronal factor that participates in the pathological processes connected with canine IBD. The observed changes probably result from the neuroprotective and/or adaptive properties of VIP. Protective and adaptive reactions induced by inflammation aim to protect the GI tract against damage by proinflammatory factors and ensure the homeostasis in the enteric nervous system (ENS) under the conditions changed by the disease process.

Changes in the expression of substance P in nerve fibres of the colonic mucosa in dogs suffering from inflammatory bowel disease.

Due to its difficult diagnosis and complicated treatment, inflammatory bowel disease (IBD) in dogs is a challenge for the veterinarian. Several aspects connected with pathological changes during IBD still remain unknown. Since one of these aspects is the participation of intestinal innervation in the evolution of the disease, the aim of this study was to demonstrate changes in the number and distribution of intramucosal colonic nerve fibres immunoreactive to substance P (SP) arising as the disease progresses. SP is one of the most important neuronal factors in intestinal innervation which, among other tasks, takes part in the conduction of pain stimuli. Using routine immunofluorescence technique, the density of nerve fibres containing SP was evaluated within mucosal biopsy specimens collected from the descending colon of healthy dogs and animals suffering from IBD of varying severity. The results of the study indicate that during severe IBD the number of nerve fibres containing SP located in the colonic mucosal layer increases in comparison to control animals. The number of SP-positive intramucosal nerves amounted to 10.99 ± 2.11 nerves per observation field in healthy dogs, 14.62 ± 2.86 in dogs with mild IBD, 14.80 ± 0.91 in dogs with moderate IBD and 19.03 ± 6.11 in animals with severe IBD. The observed changes were directly proportional to the intensity of the disease process. These observations may suggest a role of this neuronal substance in pathological processes occurring during IBD. Although the exact mechanism of the observed changes has not been completely explained, the results obtained in this investigation may contribute to improving the diagnosis and treatment of this disease, as well as the staging of canine IBD in veterinary practice.

The T2 Toxin Produced by Fusarium spp. Impacts Porcine Duodenal Nitric Oxide Synthase (nNOS)-Positive Nervous Structures-The Preliminary Study.

T2 toxin synthetized by Fusarium spp. negatively affects various internal organs and systems, including the digestive tract and the immune, endocrine, and nervous systems. However, knowledge about the effects of T2 on the enteric nervous system (ENS) is still incomplete. Therefore, during the present experiment, the influence of T2 toxin with a dose of 12 µg/kg body weight (b.w.)/per day on the number of enteric nervous structures immunoreactive to neuronal isoform nitric oxide synthase (nNOS-used here as a marker of nitrergic neurons) in the porcine duodenum was studied using the double immunofluorescence method. Under physiological conditions, nNOS-positive neurons amounted to 38.28 ± 1.147%, 38.39 ± 1.244%, and 35.34 ± 1.151 of all enteric neurons in the myenteric (MP), outer submucous (OSP), and inner submucous (ISP) plexuses, respectively. After administration of T2 toxin, an increase in the number of these neurons was observed in all types of the enteric plexuses and nNOS-positive cells reached 46.20 ± 1.453% in the MP, 45.39 ± 0.488% in the OSP, and 44.07 ± 0.308% in the ISP. However, in the present study, the influence of T2 toxin on the intramucosal and intramuscular nNOS-positive nerves was not observed. The results obtained in the present study indicate that even low doses of T2 toxin are not neutral for living organisms because they may change the neurochemical characterization of the enteric neurons.

Cyclooxygenase-2 as a Biomarker with Diagnostic, Therapeutic, Prognostic, and Predictive Relevance in Small Animal Oncology.

In canine and feline populations, the number of neoplasm cases continues to increase around the world. Attempts are being made in centres of research to identify new biomarkers that speed up and improve the quality of oncological diagnostics and therapy in human and animal tumour patients. Cyclooxygenase-2 (COX-2) is a promising biomarker with increasing relevance to human oncology, but as yet with less application in veterinary oncology. The expression of COX-2 increases significantly during pathological processes involving inflammation, pain or fever. It is also overexpressed in humans presenting various types of tumours and in selected types of tumours in animals, particularly in dogs. This article discusses the expression of COX-2 in canine and feline tumours, the importance of COX-2 as a biomarker with diagnostic, therapeutic, prognostic and predictive relevance in oncology, and the clinical significance of inhibiting COX-2 overexpression in tumours.

Long-term treatment with naproxen changes the chemical coding of the porcine intramural duodenum neurons.

Due to numerous therapeutic applications and high availability, non-steroidal anti-inflammatory drugs (NSAIDs) are the most widely used drugs worldwide. However, long-term use of these drugs can lead to damage to the gastrointestinal mucosa. The enteric nervous system (ENS), which is part of the autonomic nervous system, controls most aspects of gastrointestinal activity. Enteric neurons are characterized by considerable chemical plasticity and the appearance of a pathological factor results in a change in the synthesis of neurotransmitters. The purpose of this study was to determine the effects of naproxen on expression of biologically active substances by intramural neurons supplying the porcine duodenum. The study was performed on eight immature pigs of the Pietrain x Duroc race (approximately 20kg of body weight). The animals were divided into two groups - a control (C group) and an experimental group (N group). Group C (n=4) consisted of animals which received empty gelatine capsules. Group N (n=4) was composed of pigs who received naproxen orally for 28 days, approximately one hour before feeding. After this time, animals from both groups were euthanized. Frozen sections (14µm thickness) were then prepared from the collected duodenum and subjected to double immunofluorescence staining. Antibodies against the neuronal marker PGP 9.5 and against vasoactive intestinal polypeptide (VIP), substance P (SP), neuronal nitric oxide synthase (nNOS), galanin (GAL), pituitary adenylate cyclase-activating polypeptide (PACAP) and cocaine- and amphetamine- regulated transcript peptide (CART) were used as primary antibodies. The polyclonal donkey anti-rabbit, anti-mouse and anti-guinea pig IgG antibodies - Alexa Fluor 488 and 546 - were also used for staining. Analysis of the results obtained with a fluorescence microscope showed a significant increase in the number of nNOS-, VIP-, GAL-, PACAP- and CART-immunoreactive ganglionated neurons and a decrease in the number of SP-positive neurons in the myenteric and submucosal plexuses of the porcine duodenum. The obtained results indicate the participation of enteric neurotransmitters in the neuronal duodenal response to naproxen-induced inflammation.

The evaluation of three treatment protocols using oral prednisone and oral meloxicam for therapy of canine idiopathic lymphoplasmacytic rhinitis: a pilot study.

BACKGROUND: Idiopathic lymphoplasmacytic rhinitis (LPR) is a common inflammatory disorder of the nasal cavity in dogs due to unknown etiology. It is characterised by non-specific clinical signs, including nasal discharge, epistaxis and breathing problems. Diagnosis is usually based on the histopathologic identification of infiltrating plasmocytes and lymphocytes in the nasal mucosa and the exclusion of other underlying diseases. Treatment strategies include glucocorticoids, non-steroidal anti-inflammatory drugs, antibiotics and antifungal medications. The aim of this study was to evaluate the efficacy of various therapeutic protocols for managing canine lymphoplasmacytic rhinitis based on the results of clinical, endoscopic and histological examinations, and to determine the relapse rate for LPR in dogs.Twenty dogs of different breeds and both sexes, aged 1 to 14 years, were divided into four groups, each consisting of five dogs, including three experimental groups diagnosed with LPR and a control group.The dogs from the first experimental group were administered prednisone orally at 1 mg/kg/day in the first 4 weeks and 0,5 mg/kg/day in the following 2 weeks. The second group of dogs was administered meloxicam orally at 0,1 mg/kg/day in the first 3 weeks, followed by prednisone at 1 mg/kg/day in the following 2 weeks and 0,5 mg/kg/day in the last week of the treatment. The dogs from the third experimental group were administered meloxicam orally at 0,1 mg/kg/day for 6 weeks. The control group of dogs was administered empty gelatin capsules (placebo) orally for 6 weeks. Clinical signs, endoscopic and histopathologic lesions were scored before and after treatment. Groups were compared using Chi- squared statistics in a 2 × 2 table for pre- versus post-treatment scores. RESULTS: Clinical signs persisted in the group treated with meloxicam and were mostly resolved in prednisone-treated dogs. However, endoscopic and histological changes were still observed in these two groups after treatment. The severity of all diagnostic features was reduced in the group treated with meloxicam for 3 weeks followed by prednisone for 3 weeks. The significant differences (p < 0.05) were noted between experimental and control groups. The dogs showed a statistically significant reduction in characteristics of the LPR before and after treatment, as measured by clinical signs (Group 1vs.4 p = 0.00, group 2 vs 4 p = 0.00, group 3 vs 4 p = 0,01), by endoscopy (1 vs 4 p = 0,01, 2 vs 4 p = 0,00, 3 vs 4 p = 0,03), and by histopathology (groups 1 vs 4 p = 0,00, 2 vs 4 p = 0,00, 3 vs 4 p = 0,03). The significant differences were noted between experimental groups, as measured by endoscopy (group 2vs 3 p = 0,04), and by relapse rate (groups 1 and 2 p = 0,03, groups 2 and 3 p = 0,01). CONCLUSIONS: The three treatment protocols administered to dogs improved clinical, endoscopic and histological status. However, oral administration of meloxicam for 3 weeks, followed by prednisone for 3 weeks, appeared to be the most successful treatment. These patients remained asymptomatic for 6 months.

Inflammatory bowel disease affects density of nitrergic nerve fibers in the mucosal layer of the canine gastrointestinal tract.

The objective of this study was to determine the effect of inflammatory bowel disease (IBD) on the density of nitrergic nerve fibers in the mucosal layer of different sections of the gastrointestinal tract of dogs. Twenty-eight German shepherd hybrid dogs of both sexes, weighing from 15 to 25 kg and aged 6 to 10 y, were studied. The dogs were divided into 4 groups with 7 animals in each group: healthy animals, as well as dogs suffering from mild, moderate, and severe IBD. Immunoreactivity to neuronal isoform of nitric oxide synthase, which is a marker of nitrergic neurons, in samples of the mucosal layer in the duodenum, jejunum, and descending colon was studied using the single immunofluorescence method and the number of nerve fibers was evaluated in each observation field. The obtained results showed that IBD causes an increase in the number of nitrergic nerve fibers in all intestinal segments studied and these changes are directly proportional to the intensity of the disease process. These observations may be useful in diagnostic evaluation of the stage of canine inflammatory bowel disease in veterinary practice. The pathological mechanisms of these observed changes and the specific reasons for them are still not completely explained, however, and further study is required.

L'objectif de la présente étude était de déterminer l'effet de la maladie inflammatoire de l'intestin (MII) sur la densité des fibres nerveuses nitrergiques dans la couche muqueuse de différentes sections du tractus gastro-intestinal de chiens. Vingt-huit chiens de race Berger Allemand croisée des deux sexes, pesant entre 15 et 25 kg et âgés de 6 à 10 ans, ont fait partie de l'étude. Les chiens ont été répartis en quatre groupes de 7 chiens dans chaque groupe : des animaux en santé, aussi bien que des chiens souffrant de MII légère, modérée et sévère. L'immunoréactivité à un isoforme neuronal de l'oxyde nitrique synthase, qui est un marqueur des neurones nitrergiques, dans des échantillons de la couche muqueuse du duodénum, jéjunum, et côlon descendant a été étudiée en utilisant la méthode d'immunofluorescence unique et le nombre de fibres nerveuses a été évalué dans chaque champs observé. Les résultats obtenus montrent que la MII cause une augmentation des fibres nerveuses nitrergiques dans tous les segments intestinaux étudiés et que ces changements sont directement proportionnels à l'intensité du processus pathologique. Ces observations pourraient être utiles pour l'évaluation diagnostique des stades de la MII en pratique vétérinaire. Par contre, les mécanismes pathologiques des changements observés et leurs raisons spécifiques ne sont pas encore complètement élucidés et des études supplémentaires sont requises.(Traduit par Docteur Serge Messier).

Effects of inflammation and axotomy on expression of acetylcholine transferase and nitric oxide synthetase within the cocaine- and amphetamine-regulated transcript-immunoreactive neurons of the porcine descending colon.

This study reports changes in expression of acetylcholine transferase (AChT) and nitric oxide synthetase (NOS) in neurons immunoreactive for cocaine- and amphetamine-regulated transcript (CART) peptides during chemically-driven inflammation and axotomy in the porcine descending colon. The co-localization of the neurotransmitters with CART was studied by double immunofluorescence in the myenteric plexus (MP) and outer submucosal plexus (OSP) of the porcine descending colon under physiological and selected pathological conditions. In control animals, neurons expressing CART also expressed AChT in 25.37 ± 0.98% and 26.73 ± 0.96% in the MP and OSP, respectively. Neuronal co-expression of CART with NOS occurred in 90.66 ± 2.13% and 88.09 ± 2.96% in the MP and OSP, respectively. Following axotomy the number of neurons co-expressing CART and AChT decreased to 16.50 ± 3.20% in the MP and increased to 35.49 ± 2.04% in the OSP, while the number of neurons co-expressing CART and NOS increased to 96.66 ± 2.38% in the MP and 97.46 ± 2.22% in the OSP. Experimentally-induced colitis resulted in an increase in the number of neurons co-expressing CART and AChT to 42.40 ± 2.28% in the MP and 63.62 ± 1.83% in the OSP. Similarly, in these animals the number of neurons co-expressing CART and NOS increased to 93.9 ± 2.58% in the MP and 90.43 ± 2.09% in the OSP. Sham-operated controls showed expression levels of 26.22 ± 0.66% (MP) and 27.02 ± 1.73% (OSP) for simultaneous CART and AChT expression and 94.18 ± 0.93% (MP) and 88.21 ± 0.81% (OSP) for CART and NOS co-localization. These data confirm that the examined neurotransmitters have a role in traumatic and inflammatory responses of enteric neurons.

The effectiveness of natural and synthetic immunomodulators in the treatment of inflammatory bowel disease in dogs.

The aim of the study was to evaluate the usefulness of immunomodulators in the treatment of inflammatory bowel disease (IBD) in dogs. Twenty-eight dogs diagnosed with IBD took part in the study. The animals received a food containing extruded immunomodulators: ß-1,3/1,6-D-glucan, ß-hydroxy-ß-methyl-butyrate (HMB) and levamisole for 42 days. Whole blood samples were analysed before and after therapy assessing changes in phagocyte activity (respiratory burst activity, RBA and potential killing activity, PKA), evaluation of proliferation response of mitogen-stimulated lymphocytes and serum gamma globulin levels, lysozyme activity, ceruloplasmin levels and interleukin activity (IL-6 and IL-10). In this experiment, ß-1,3/1,6-D-glucan delivered the highest level of treatment efficacy by producing the quickest therapeutic effect, lowering Canine Inflammatory Bowel Disease Activity Index (CIBDAI) values to below 3, improving histopathological parameters, decreasing IL-6 levels, increasing IL-10 concentrations, and producing remission periods longer than six months. HMB and levamisole were also effective in lowering CIBDAI scores, but the abatement of clinical symptoms was slower and less pronounced in comparison with ß-1,3/1,6-D-glucan. The results indicate that ß-1,3/1,6-D-glucan can be useful in the treatment of canine IBD.