Molecular identification of bacteria associated with canine periodontal disease

Hsp90 is a molecular chaperone that is involved in diverse cellular processes including protein folding/repairing and signal transduction. Edwardsiella tarda is a serious fish pathogen that affects fish aquaculture worldwide. The aim of this study was to investigate the potential importance of HtpG, the prokaryotic homologue of Hsp90, in the pathogenesis of E. tarda. E. tarda HtpG is 627-residue in length and contains domain structures that are conserved among Hsp90 family members. Quantitative real time RT-PCR analysis indicated that expression of htpG is induced by heat shock and oxidative stress. Recombinant HtpG (rHtpG) purified from Escherichia coli exhibits apparent ATPase activity, which is optimal at 40°C. Mutation of htpG (i) affects bacterial growth at elevated temperature and renders the cells more sensitive to stress induced by reactive oxygen species, (ii) causes dramatic reduction in blood dissemination and general bacterial virulence, (iii) weakens the ability of E. tarda to block head kidney macrophage activation and to resist against the bactericidal effect of macrophages, and (iv) upregulates the expression of pro-inflammatory cytokines in macrophages. Taken together, these results indicate that HtpG is a biologically active protein that is required for E. tarda to cope with various stress conditions especially that encountered in vivo the host system during infection.     

Systemic effects of periodontal disease in cats

Background: Periodontal disease in cats is a local disease that may have systemic consequences that are affected by treatment.

Objective: To test the hypotheses that systemic health indices would be correlated with the severity of periodontitis, and would improve with treatment.

Animals and methods: Apparently otherwise healthy cats from an in-bred colony were randomly assigned to a treatment group (n?=?30), or a control group (n?=?18), which was left untreated for 3 months. Periodontal disease was scored at baseline in the treatment group according to calculus, gingivitis, and alveolar bone loss measured from dental radiographs. Blood, urine and saliva were collected from both groups before, and 16, 45, and 90 days after dental treatment. Assays included haematology, urinalysis, serum biochemistry, serum IgG, salivary IgA, lymphocyte subsets and proliferation, and plasma malonyldialdehyde (MDA). Correlations between the severity of periodontitis and assays at baseline were assessed, and the effect of treatment determined using linear mixed model methodology.

Results: The severity of periodontitis was associated with age, bodyweight, total globulins (Globs), Alanine aminotransferase, and IgG, and negatively associated with albumin, haemoglobin, haematocrit, and Aspartate aminotransferase (AST). Treatment significantly reduced IgG, total Globs, AST, and eosinophils, and increased cholesterol. Other leucocyte assays and plasma MDA concentrations were not affected by the treatment. Cats ate dry food faster 1 week after, than they did 1 week before treatment.

Conclusions and clinical relevance: Although the clinical significance of these findings are unknown, we conclude that periodontitis is not simply a localized disease, but also impacts on systemic health and wellbeing.     

Comparative palatability of five supplements designed for cats suffering from chronic renal disease

Background

Intestinal phosphate binders, uremic toxin binders and some other types of supplements are an integral part of the management of chronic kidney disease (CKD) in various species, including cats. This pathology in domestic carnivores requires life-long nutritional and medical management. In this context, the compliance of owners and patients cannot be achieved without an adequate level of palatability for oral medication or supplementation. Knowing that hyporexia and anorexia are among the most commonly seen clinical signs in cats suffering from CKD this is already, in itself, a serious obstacle to acceptable compliance in sick animals. The aim of the present study was to investigate the palatability of four commercially available products designed for cats suffering from CKD: Ipakitine® (Vetoquinol, France), Azodyl® (Vetoquinol, USA), Renalzin® (Bayer, France), Rubenal® (Vetoquinol, France) and an additional recently developed product: Pronefra® (Virbac, France). The study was performed with a group of previously-characterised cats, all living in an enriched and well-being securing environment of an independent centre housing panels of pets expert in palatability measurement. In total 172 monadic testings were performed. The palatability of each product was assessed by measuring their rates of prehension and consumption, and the consumption proportions were also analysed.

Results

The most palatable presentation (based on useful consumption) was Pronefra®, which was significantly higher than Azodyl® (p = 0.046), Ipakitine® (p < 0.0001), Renalzin® (p < 0.0001) and Rubenal® (p < 0.0001). The product with the highest rate of prehension was also Pronefra®, which was significantly higher than Azodyl® (p = 0.0019), Ipakitine® (p = 0.0023), Renalzin® (p = 0.0008) and Rubenal® (p < 0.0001).

Conclusion

Pronefra® was the most palatable presentation tested, meaning it may be useful for improving ease of supplementation in CKD cats.     

Molecular identification of Entamoeba histolytica from stool samples of Ilam,Iran

IntroductionAmoebiasis is a multifactorial, life-threatening public health issue and the third parasitic disease cause of mortality in worldwide, particularly in low- and mid-income countries. The aim of this study was to attempt to explore genetic encoding differences of CP8 (conserved gene) of Entamoeba histolytica/Entamoeba dispar in its various infectious properties isolated from Ilam located at a southwest part of Iran.Materials and methodsA total of 2023 stool samples were collected between 2016 and 2018 from the hospital in Ilam, of which only 30 isolates were identified as E. histolytica/E. dispar. These isolates were collected from the intensive care unit, infectious disease, and surgery settings. The isolates were identified and the polymerase chain reaction (PCR) was performed to detect the CP8 gene. In all stages, Entamoeba histolytica HM1: IMSS was used as a positive control.ResultsIn genotype confirmation, only two isolates had the CP8 gene found in the PCR technique. The sequencing results confirmed the mentioned gene with 99%–100% specificity.ConclusionIt is concluded that PCR is highly sensitive to detect E. histolytica and indicating this important role as screening tools in direct DNA extraction from stool samples and valuable technique in early detection of symptomatic and asymptomatic E. histolytica patients.     

Preliminary study of the anaerobic bacteria isolated from subgingival plaque from sheep

A preliminary study was made to determine the genera of cultivable anaerobic bacteria which could be isolated from subgingival plaque of sheep. Samples were taken from 10 sheep on farms with a known record of broken mouth periodontitis. For assessment of the sampling technique, samples were also taken from freshly exposed tooth roots in killed sheep. The bacteria isolated on several selective and non-selective media were identified to genus level by a combination of Gram reaction, colony morphology and gas chromatographic analysis of volatile and non-volatile fatty acid metabolic end products. At least 10 different genera were isolated and these findings are discussed in relation to the bacteriology of human periodontitis and recent studies of sheep broken mouth periodontitis.     

Chromosomal DNA probes for the identification of asaccharolytic anaerobic pigmented bacterial rods from the oral cavity of cats.

A dot-blot hybridisation assay using isolated high molecular weight DNA as whole chromosomal probes of the cat pigmented asaccharolytic Bacteroides/Porphyromonas species was used against both purified high molecular weight DNA and DNA released on membranes from whole cells for the identification of B. salivosus and for its differentiation from the other anaerobic species isolated from normal and diseased mouths of cats and horses. 32P-labelled probes were compared with digoxigenin (DIG)-labelled probes (Boehringer-Mannheim). The whole chromosomal probes were specific--differentiating B. salivosus from a variety of species (including members of the genera Bacteroides, Fusobacterium, Eubacterium, and Prevotella) found in normal and abnormal mouths of cats and horses. Likewise, asaccharolytic black pigmented Group 2 strains were distinguishable from all strains tested. However, cat strains of P. gingivalis which show 68-76% DNA-DNA homology with human strain P. gingivalis ATCC 33277T, were not distinguishable from each other using either 32P-labelled or DIG-labelled probes. The minimum amount of pure Bacteroides DNA which could be detected by the 32P-labelled probe was 100-300 pg, while the amount of pure DNA detected by the DIG system was 1-3 mg after room temperature colour development for 1 h and 100-300 pg after 6 h colour development.     

Molecular detection of Bartonella henselae and Bartonella clarridgeiae in clinical samples of pet cats from Southern Italy

Bartonella henselae is considered an emerging pathogen of veterinary and medical interest that can be occasionally transmitted to humans. Cats are considered to be the only reservoir host for B. henselae. In this study, we used a nested-PCR assay to investigate the prevalence of B. henselae and Bartonella clarridgeiae DNA in peripheral blood samples, fine needle lymph node aspirate specimens and oral swabs from 85 cats in order to develop an easy diagnostic strategy for the selection of infection-free cats that are being considered as pets, especially for immunocompromised patients. Overall, molecular analysis showed that 71 cats (83.5%) tested PCR positive for the presence of B. henselae DNA. PCR amplification of DNA B. henselae produced positive products from lymph node aspirate specimens (62/85; 72.9%) similar to those obtained from blood samples (60/85; 70.6%) and higher than those from oral swabs (51/85; 60%) of cats. No PCR product was obtained for B. clarridgeiae. The simultaneous analysis of three different clinical samples in our study increased the diagnostic possibilities for B. henselae infection in the examined cats from 60–72.9% to 83.5%. Lymph node aspirates were found to be the most effective clinical samples for the detection of B. henselae and blood samples were the next best. Oral swab samples were used in this study with good results when considered in combination with blood and/or lymph node aspiration. The use of nested-PCR assay on these three clinical samples may enhance the diagnostic sensitivity for bartonellosis in cats irrespective of the clinical status of animals.     

Associations amongst three feline Porphyromonas species from the gingival margin of cats during periodontal health and disease

Digoxigenin labelled whole chromosomal DNA probes directed against three feline members of the genus Porphyromonas (P. gingivalis VPB 3492, P. circumdentaria NCTC 12469T and P. salivosa VPB 3313) were used to identify and quantify organisms in samples taken from the gingival margins of 40 domestic cats with different grades of periodontal disease. At the right upper canine tooth, the grade of periodontal disease ranged from 0 to 5 and the cfu of facultative/obligate anaerobes ranged from 5.5 x 10(4) to 2.0 x 10(6)). In 38 of the 40 cats, at least one of the three Porphyromonas species was isolated and regression analysis showed that the cfu of total Porphyromonas sp. was a highly significant indicator of the grade of periodontal disease (p < 0.001, R2 0.510). Feline P. gingivalis was isolated from 37 of the 40 cats and regression analysis showed that it was a highly significant predictor of the grade of periodontal disease (p < 0.001, R2 0.561). The cfu of P. salivosa was a significant predictor of the grade of periodontal disease (p < 0.001, R2 0.286) and regression analysis showed that there was a significant positive relationship between cfu of P. circumdentaria and grade of periodontal disease (p = 0.018, R2 0.116). The periodontal grades at the right upper third premolar tooth ranged from 0 to 6. The cfu of facultative/obligate anaerobes isolated ranged from 1.2 x 10(5) to 7.9 x 10(6), and regression analysis showed that cfu was a significant predictor of periodontal grade (p < 0.001, R2 0.378). The cfu of total Porphyromonas species ranged from 1.2 x 10(4) to 1.7 x 10(6) and regression analysis of the cfu against the grade of periodontal disease showed a highly significant association (p < 0.001, R2 0.633). The cfu of P. gingivalis ranged from 0 to 1.1 x 10(6) and regression analysis of the cfu of P. gingivalis against the grade of periodontal disease showed a highly significant association (p < 0.001, R2 0.439). The cfu of P. salivosa was a significant predictor of the grade of periodontal disease (p < 0.001, R2 0.479) and the same association was found between cfu of P. circumdentaria and grade of periodontal disease (p = 0.002, R2 0.204). This study has established Porphyromonas as anumerically significant and highly prevalent genus in feline periodontal disease.     

Long‐term use of non‐steroidal anti‐inflammatory drugs in cats with chronic kidney disease: from controversy to optimism

This is the first of a series of capsule reviews published by the World Small Animal Veterinary Association ‐ Global Pain Council (WSAVA‐GPC). Each of these short articles provides a brisk assessment of the scientific evidence in specific aspects of pain management, including analgesic techniques, recommendations and controversies surrounding their use. In this first capsule review, the scientific evidence available on the long‐term use of non‐steroidal anti‐inflammatory drugs in cats with concomitant chronic pain and chronic kidney disease is discussed.     

Detection of singlet oxygen in blood serum samples of clinically healthy lambs and lambs suffering from alveld disease

Alveld is a disease in lambs of domestic sheep (Ovis aries L.), characterized by a combination of photosensitivity and liver damage. Generation of singlet oxygen play a major role in phototoxicity reactions. The compound phylloerythrin (phytoporphyrin) is so far assumed to be the main photodynamic agent in hepatogenous photosensitivity diseases in sheep. Phylloerythrin is a potent photosensitizer and an efficient source of singlet oxygen. The compound accumulates in the peripheral circualtion upon liver damage. Liver dysfunction is also likely to cause an increase in the blood level of bilirubin. Formation of singlet oxygen by bilirubin is reported. In the present work the photosensitizing potential of serum has been measured and related to the bilirubin- and phylloerythrin levels in lambs suffering from alveld and in clinically healthy controls. The singlet oxygen level of the serum was taken as a measure of the photosensitizing potential. The observed singlet oxygen values in serum from alveld lambs were significantly higher than the corresponding values observed in clinically healthy control lambs. This indicates that the serum of the alveld lambs contains an elevated concentration of photosensitizer. The singlet oxygen level was not correlated to the concentration of bilirubin or phylloerythrin. The results indicate that the photosensitizing mechanism is quite complex and may involve other sensitizer(s) than phylloerythrin.     

Efficacy of a commercial hydrolysate diet in eight cats suffering from inflammatory bowel disease or adverse reaction to food

Eight of 28 cats presenting with chronic vomiting and/or diarrhoea were diagnosed as suffering from chronic enteropathy. All cats had undergone a complete diagnostic work-up, including endoscopy. Histopathology findings varied from normal, eosinophilic, to lymphoplasmacytic gastroenteritis. On the basis of findings, the cats were diagnosed with inflammatory bowel disease (IBD) or an adverse reaction to food. A hydrolysed protein diet was used as sole therapy, and clinical signs resolved within 4-8 days in all eight cats. After 2 months the cats had gained weight. A challenge trial with their previous diet resulted in recurrence of the clinical signs, which resolved in seven cats when the test diet was reintroduced. We conclude that the hydrolysed protein diet used in the current study was effective in managing these cases.     

Molecular characterization of Malassezia nana isolates from cats

Malassezia nana (M. nana) is a lipid-dependent yeast that has been isolated from cats and cows. Some sequence variability has been observed in the large subunit (LSU) and internal transcribed spacer (ITS) regions between strains isolated from cats and cows though these regions in M. nana isolates from cats alone have proven to be relatively conserved. In the present study, microsatellite PCR fingerprinting and β-tubulin gene sequence analysis were carried out on M. nana isolates from cats to investigate the genetic diversity of this species. Although a relatively small number of isolates were available, the similarity in the sequences of the β-tubulin and the microsatellite profiles indicate that a particular M. nana genotype colonizes cats. Moreover, all isolates obtained from animals with otitis externa had the same microsatellite fingerprinting pattern. Further studies of a wider population of M. nana isolates from other hosts and status disease are needed to establish that M. nana is a genetically homogeneous species. This is the first report of the characterization of the β-tubulin gene in Malassezia spp.     

Molecular characteristics of cutaneous papillomavirus from the canine pigmented epidermal nevus

To investigate the relation between the canine pigmented epidermal nevus (PEN) and cutaneous papillomavirus, we cloned and sequenced the L1 gene of papillomavirus from the canine pigmented epidermal nevus (PEN). Amplification of DNA sample with the L1 consensus primers yielded an expected fragment of approximately 450-bp. The nucleotide sequences of the fragment showed about 64% of sequence similarity to the L1 region of human papillomavirus isolate CP6108 and less than 57% sequence similarity to those of canine oral papillomavirus (COPV). In situ hybridization determined the presence of papillomavirus DNA mainly in the upper stratum granulosum of skin in this case. The results indicated that the canine cutaneous papillomavirus from the PEN lesion was genetically close to human papillomavirus rather than COPV.     

Antimicrobial drug susceptibility of bacteria isolated from disease processes in cattle, horses, dogs and cats

Results are presented of antimicrobial disc diffusion susceptibility testing on commonly isolated bacterial pathogens made at the Ontario Veterinary College Diagnostic Bacteriology Laboratory in 1981 and 1982. Nearly 2 000 isolates from horses, cattle, dogs and cats were tested. Comparison of resistance in the same bacterial species isolated from different animal species showed significant differences between some of the same antibiotics.     

猫源弓形虫虫株的分离与鉴定

将猫的心、脑、舌用盐酸-胃蛋白酶溶液消化处理后接种小白鼠，经连续传代分离弓形虫虫株，用特异PCR方法对所分离的虫株进行了鉴定。分别从24只猫的样品中分离出8株弓形虫虫株，用弓形虫种特异的PCR方法对8个虫株进行鉴定。均得到弓形虫的特异条带；测序证明所扩增出的DNA片段确为弓形虫的核糖体DNA第一 内转录间隔区（ITS1）部分序列。研究结果表明，将动物组织用盐酸-胃蛋白酶溶液消化处理后接种小白鼠是一种分离弓形虫虫株较为理想的方法．特异PCR方法能准确、快速地鉴定弓形虫虫株。     

Clinical evaluation of meloxicam versus ketoprofen in cats suffering from painful acute locomotor disorders

The aim of this study was to evaluate the efficacy and palatability of meloxicam 0.5mg/ml oral suspension, compared to ketoprofen tablets in cats suffering from painful acute locomotor disorders. This single blinded, positively-controlled, randomised, multicentre trial involved 121 client owned cats. Cats received either meloxicam (0.5mg/ml oral suspension) at 0.1mg/kg on day 1 followed by 0.05mg/kg q 24h on days 2-5, or ketoprofen 5mg tablets at 1.0mg/kg q 24h for 5 days. The efficacy of the two treatments was assessed subjectively by clinicians on day 6 using a clinical sum score (CSS). Palatability and accuracy of dosing were also assessed. The baseline CSS was not significantly different between the groups, and after 5 days of treatment the CSS had decreased to a similar extent, reflecting a reduction in pain. There were no significant differences between the CSS of each group at day 6. Both treatments were well tolerated. Meloxicam was significantly more palatable than ketoprofen, and allowed for more accurate dosing. Meloxicam and ketoprofen are a safe and efficacious treatment for acute locomotor disorders in cats. Meloxicam (Metacam) may be associated with superior compliance in clinical practice due to the higher palatability, which results in better ease of administration.     

水貂主要病原的分子生物学鉴定及多重感染的病原分析

对2012年8月至2013年12月接诊的山东地区的241份病死貂进行病原学鉴定,主要包括生物学特性分析和16S rRNA序列或特定序列的同源性分析,并对多重感染进行统计分析。结果表明,山东地区引起水貂发病的主要病原有:犬瘟热病毒(25.3%)、细小病毒(25.3%)、阿留申病毒(19.1%)、绿脓杆菌(49.0%)、克雷伯氏菌(19.9%)、大肠杆菌(11.2%)、奇异变形杆菌(8.3%)以及巴氏杆菌(5.0%)和肺炎双球菌(2.5%)等。而且主要以单独感染(46.0%)和二重感染(44.4%)为主,三重感染(7.9%)次之,四重感染(1.24%)和五重感染(0.41%)较少。通过本研究掌握了山东地区水貂多重感染的主要病原及其特性,摸清了其感染发病规律,为山东地区规模化养貂场多重感染的诊断与防制提供理论依据。