Identification of bacteria associated with feline chronic gingivostomatitis using culture-dependent and culture-independent methods

Feline chronic gingivostomatitis (FCGS) is a chronic inflammatory disease of the oral cavity that causes severe pain and distress. There are currently no specific treatment methods available and little is known regarding its aetiology, although bacteria are thought to play a major role. The purpose of this study was to identify the oral bacterial flora in normal and diseased cats. Oral swabs were obtained from the palatoglossal folds of eight cats (three normal and five FCGS) and were subjected to microbiological culture. Pasteurella pneumotropica and Pasteurella multocida subsp. multocida were the most prevalent species identified by culture methods in the normal and FCGS samples, respectively. Bacteria were also identified using culture-independent methods (bacterial 16S rRNA gene sequencing). For the normal samples, 158 clones were analysed and 85 clones were sequenced. Capnocytophaga canimorsus (10.8% of clones analysed) was the predominant species. Uncultured species accounted for 8.2% of clones analysed, and 43.7% of clones analysed represented potentially novel species. For the FCGS samples, 253 clones were analysed and 91 clones were sequenced. The predominant species was P. multocida subsp. multocida (51.8% of clones analysed). Uncultured species accounted for 8.7% of clones analysed, and 4.7% of clones analysed represented potentially novel species. It is concluded that the oral flora in cats with FCGS appears to be less diverse than that found in normal cats. However, P. multocida subsp. multocida is found to be significantly more prevalent in FCGS than in normal cats and consequently may be of aetiological significance in this disease.     

The use of oral recombinant feline interferon omega in two cats with type II diabetes mellitus and concurrent feline chronic gingivostomatitis syndrome

Feline Chronic Gingivostomatitis Syndrome (FCGS) is a common disease in clinical practice. Among the therapeutic options available, long-acting corticosteroids are frequently used due to their anti-inflammatory and immunosuppressive properties. Although they may improve the clinical symptoms, they can lead to a progressive form of the disease that becomes refractory to treatment. Furthermore, their direct relationship with type II diabetes mellitus (DM) is well known. Consequently, these drugs are controversial and not recommended for routine management of FCGS. Recombinant feline interferon-omega (rFeIFN-ω) is an immunomodulatory compound. Recently, its daily oral administration has been shown to be successful in treating refractory cases of FCGS. This case study describes two clinical cases of type II DM complicated by FCGS. Both animals were calicivirus positive and they had been previously treated with long-acting corticosteroids, which may have been the major cause of DM. The two cats were treated with glargine insulin (Lantus, starting dose 1 IU/cat twice daily (BID)), achieving remission 10 and 18 weeks later respectively. Considering the difficulty with control of FCGS in these animals, an oral daily dose of rFeIFN-ω was started as an alternative to long-acting corticosteroids. In both cats oral clinical signs gradually improved and 60 days after the start of therapy the owners reported a significant relief of pain during mastication. According to the authors’ knowledge, this is the first case report that describes the successful use of rFeIFN-ω in the management of FCGS in type II diabetic cats, in which long-acting corticosteroids are contraindicated.     

Relevance of feline calicivirus, feline immunodeficiency virus, feline leukemia virus, feline herpesvirus and Bartonella henselae in cats with chronic gingivostomatitis

Despite its common occurrence, the aetiology of chronic gingivostomatitis in cats remains uncertain. Aetiology is likely multifactorial, and several infectious agents may be associated with chronic gingivostomatitis. The purpose of this study was to investigate the prevalence of feline calicivirus (FCV), feline immunodeficiency virus (FIV), feline leukemia virus (FeLV), feline herpesvirus (FHV), and Bartonella henselae (B. henselae) in cats with chronic gingivostomatitis and in an age-matched control group. In addition, other factors, e. g., environmental conditions were investigated. In 52 cats with chronic gingivostomatitis and 50 healthy age-matched control cats, the presence of FCV ribonucleic acid (RNA), and FHV deoxyribonucleic acid (DNA) (polymerase chain reaction [PCR] from oropharyngeal swabs), and B. henselae DNA (PCR from oropharyngeal swabs and blood), as well as FeLV antigen (serum), and antibodies against FCV, B. henselae, and FIV (serum) were examined. FCV RNA was significantly more common in cats with chronic gingivostomatitis (53.8%, p < 0.001) than in controls (14.0%); a significant difference was also found in the prevalence of antibodies to FCV between the cats with chronic gingivostomatitis (78.8%, p = 0.023) and controls (58.0%). Of the other infectious agents investigated, there was no significant difference in the prevalence between the cats with chronic gingivostomatitis and the controls. The results of this study allow the conclusion that FCV, but no other infectious agents, is commonly associated with chronic gingivostomatitis in cats.     

Salivary and serum immunoglobulin levels in cats with chronic gingivostomatitis

The salivary and serum concentrations of immunoglobulins G, M and A (IgG, IgM and IgA), and the salivary concentrations of albumin were measured by ELISA in 30 cats with chronic gingivostomatitis and 32 healthy cats. The cats with chronic gingivostomatitis had significantly higher salivary concentrations of IgG, IgM and albumin, and higher serum concentrations of IgG, IgM and IgA, but significantly lower salivary concentrations of IgA than the healthy cats. The cats with chronic gingivostomatitis were treated with either methylprednisolone, sodium aurothiomalate, metronidazole and spiramycin, or oral hygiene products. After three months of treatment, the cats receiving methylprednisolone had a significant reduction in serum IgG levels compared to the cats treated with sodium aurothiomalate or metronidazole and spiramycin, but after six months of treatment there were no significant differences between the groups. Before the treatments, the levels of oral inflammation were not correlated significantly with any of the serum or salivary immunoglobulin levels. However, the changes in oral inflammation were correlated significantly with the changes in the salivary IgM concentration after three and six months of treatment, and with the change in the salivary IgA concentration after six months of treatment.     

Cessation of feline calicivirus shedding coincident with resolution of chronic gingivostomatitis in a cat

Feline calicivirus (FCV) shedding and oral bacterial flora were monitored over a period of 22 months in a case of feline gingivostomatitis (FGS). The cat was treated daily with 50 mg thalidomide capsules by mouth, and 200 mg lactoferrin powder was applied directly to the lesions. Clinical signs began to resolve after 11 months when, in addition to treatment, the diet had been changed to an additive-free cat food supplemented with antioxidant vitamins A, D3 and E. Resolution of clinical signs of FGS coincided with the cessation of FCV shedding, and this is the first report documenting such an association. Which part of the treatment, if any, contributed to the cure requires further investigation.     

荧光定量RT-PCR检测雏鸡感染NDV强毒后胸腺和法氏囊中TLR2与TLR4 mRNA的表达

Toll样受体2(TLR2)和Toll样受体4(TLR4)在识别病原微生物过程中发挥重要作用。为了定量检测TLR2、TLR4 mRNA表达水平,研究病原与机体的相互作用,本研究建立了检测鸡TLR2、TLR4 mRNA表达水平的SYBR GreenⅠ荧光定量RT-PCR(RRT-PCR)方法,检测了新城疫病毒强毒(vNDV)感染SPF雏鸡后36h、48h时胸腺、法氏囊中TLR2、TLR4 mRNA表达量变化。结果显示该方法特异性好,RRT-PCR产物分别在85.5℃、83.3℃出现单特异峰,对TLR2、TLR4的扩增效率分别为105.91%和95.30%,相关系数分别为0.9980、0.9996,最低检测限分别为108拷贝/反应和461拷贝/反应。感染后36h vNDV显著抑制TLR2、TLR4基因在法氏囊、胸腺中的表达;感染后48h时,法氏囊、胸腺中TLR2基因的表达水平显著升高,胸腺中TLR4基因的表达显著升高,而法氏囊中TLR4基因的表达仍处于抑制状态。本研究证明TLR2和TLR4参与了鸡体对NDV的感染应答。     

Toll-like receptor and pro-inflammatory cytokine expression during prolonged hyperinsulinaemia in horses: Implications for laminitis

Equine laminitis, a disease of the lamellar structure of the horse's hoof, can be incited by numerous factors that include inflammatory and metabolic aetiologies. However, the role of inflammation in hyperinsulinaemic laminitis has not been adequately defined. Toll-like receptor (TLR) activation results in up-regulation of inflammatory pathways and the release of pro-inflammatory cytokines, including interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α), and may be a pathogenic factor in laminitis. The aim of this study was to determine whether TLR4 expression and subsequent pro-inflammatory cytokine production is increased in lamellae and skeletal muscle during equine hyperinsulinaemia. Standardbred horses were treated with either a prolonged, euglycaemic hyperinsulinaemic clamp (p-EHC) or a prolonged, glucose infusion (p-GI), which induced marked and moderate hyperinsulinaemia, respectively. Age-matched control horses were treated simultaneously with a balanced electrolyte solution. Treated horses developed clinical (p-EHC) or subclinical (p-GI) laminitis, whereas controls did not. Skeletal muscle and lamellar protein extracts were analysed by Western blotting for TLR4, IL-6, TNF-α and suppressor of cytokine signalling 3 (SOCS3) expression. Lamellar protein expression of TLR4 and TNF-α, but not IL-6, was increased by the p-EHC, compared to control horses. A significant positive correlation was found between lamellar TLR4 and SOCS3. Skeletal muscle protein expression of TLR4 signalling parameters did not differ between control and p-EHC-treated horses. Similarly, the p-GI did not result in up-regulation of lamellar protein expression of any parameter. The results suggest that insulin-sensitive tissues may not accurately reflect lamellar pathology during hyperinsulinaemia. While TLR4 is present in the lamellae, its activation appears unlikely to contribute significantly to the developmental pathogenesis of hyperinsulinaemic laminitis. However, inflammation may have a role to play in the later stages (e.g., repair or remodelling) of the disease.     

The canine and feline skin microbiome in health and disease

The skin harbours a diverse and abundant, yet inadequately investigated, microbial population. The population is believed to play an important role in both the pathophysiology and the prevention of disease, through a variety of poorly explored mechanisms. Early studies of the skin microbiota in dogs and cats reported a minimally diverse microbial composition of low overall abundance, most probably as a reflection of the limitations of testing methodology. Despite these limitations, it was clear that the bacterial population of the skin plays an important role in disease and in changes in response to both infectious and noninfectious diseases. Recent advances in technology are challenging some previous assumptions about the canine and feline skin microbiota and, with preliminary application of next‐generation sequenced‐based methods, it is apparent that the diversity and complexity of the canine skin microbiome has been greatly underestimated. A better understanding of this complex microbial population is critical for elucidation of the pathophysiology of various dermatological (and perhaps systemic) diseases and to develop novel ways to manipulate this microbial population to prevent or treat disease.     

口服盐酸特比萘芬对猫肝肾损伤的病理组织学观察

选取健康猫28只,饲喂特比萘芬片剂,将实验猫随机分为特比萘芬低剂量(10 mg/kg.d)、中剂量(20 mg/kg.d)、高剂量(40 mg/kg.d)和对照组(0 mg/kg.d)4个剂量组,连续给药3周。在给药后第3周(即第21天)和第5周(即第35天)取不同剂量组的实验猫各3只,安乐死。采取肝脏和肾脏组织。采用H.E.染色方法检测盐酸特比萘芬对猫肝肾的组织病理学损伤。结果显示,不同剂量组盐酸特比萘芬均能造成不同程度的猫肝肾实质病变,且盐酸特比萘芬对肝肾的损伤具有明显的正向量效关系;且连续给药3周后停药2周时所造成的猫肝肾的损伤比在连续给药3周期间所造成的损伤严重。     

Expression of Toll-like receptor 9 (TLR9) in the lungs and lymphoid tissue of pigs

The pattern of distribution of Toll-like receptor 9 (TLR9) in different tissues varies between species. The aim of the present study was to describe the distribution of TLR9 expression in selected tissues and organs of healthy pigs at 3 weeks and 3 months of age. Representative formalin-fixed samples of lung, thymus and secondary lymphoid tissues were evaluated by immunohistochemistry. TLR9 positive staining was observed in epithelial cells, vascular endothelium and myoepithelial-like cells, as well as in cells of the alveolar septa of the lung. Antigen presenting cells of perifollicular zones (interdigitating, macrophage and dendritic-like cells) of the Peyer's patches, lymph nodes, spleen and thymus were also immunoreactive for TLR9. No differences were seen in TLR9 protein expression in tissues from the two age groups.