Sequence and phylogenetic analysis of the 16S rRNA gene of Ehrlichia canis strains in dogs with clinical monocytic ehrlichiosis     

Siarkou VI  Mylonakis ME  Bourtzi-Hatzopoulou E  Koutinas AF 《Veterinary microbiology》2007,125(3-4):304-312

The purpose of this study was to characterize, at the molecular level, the Ehrlichia canis strains involved in naturally occurring canine monocytic ehrlichiosis (CME) in Greece, and to investigate if any sequence diversity exists between the 16S rRNA genes of those involved in the mild non-myelosuppressive or the severe myelosuppressive form of CME. To this end, amplification of the ehrlichial 16S rRNA gene was attempted by nested polymerase chain reaction (PCR) assays in bone marrow (BM) aspirates from 20 dogs tentatively diagnosed as having non-myelosuppressive (n=10, group A) or myelosuppressive (n=10, group B) CME. PCR assay using E. canis-specific primers revealed that 15 BM samples, including all group A and 5 group B dogs, were positive. Using universal PCR primers, a nearly full-length 16S rRNA gene could be amplified from 13 BM samples, including 9 group A and 4 group B dogs. The 16S rDNA analysis based on secondary structure revealed that all sequences of the Greek strains were identical to each other and indicated 100% identity among some American (Venezuelan and Brazilian), European (Greek), Middle Eastern (Turkish) and Asiatic (Thailand) strains. The results of this study suggest that the E. canis strains involved in the non-myelosuppressive and myelosuppressive forms of CME in Greece share an identical 16S rRNA genotype.  相似文献   

Clinical, histopathological and therapeutic considerations in a flock of sheep with facial staphylococcal-associated dermatitis     

Koutinas AF  Saridomichelakis MN  Argyroudis S  Koutinas CK  Karatzanos P  Giadinis N 《Veterinary dermatology》2007,18(4):211-216

In a flock of 290 sheep, a total of 20 ewes (6.9%) with an age range of 1.5-4 years, developed a nonpruritic dermatitis characterized by alopecia/hypotrichosis, erythema, hyperpigmentation, crusting, superficial ulcers, exudation and thickening of the skin that was localized mainly to the peri-ocular area. Frequent and massive feeding of swarming sucking flies on the lesional sites was witnessed. In every instance (n = 6), Staphylococcus aureus was cultured from the exudate and the aseptically obtained skin biopsy samples from both recurrent and new cases. The main predisposing factor was the presence of small troughs, which forced the sheep to compete for food. Intramuscular administration of cefalexin for 7 days resulted in the resolution of lesions over a 2-week period. No further cases or recurrences were seen over the 16-month follow-up period. The main histopathological feature of this skin disease was a superficial and deep, perivascular to interstitial eosinophilic and mononuclear dermatitis.  相似文献   

A randomised, blinded, placebo-controlled clinical trial with allopurinol in canine leishmaniosis.     

A F Koutinas  M N Saridomichelakis  M E Mylonakis  L Leontides  Z Polizopoulou  C Billinis  D Argyriadis  N Diakou  O Papadopoulos 《Veterinary parasitology》2001,98(4):247-261

A total of 45 non-uremic dogs, with clinical signs indicating leishmaniosis, entered the study. Diagnosis was confirmed by indirect immunofluorescence assay (IFA) on serum and polymerase chain reaction (PCR) on bone marrow samples. The dogs were randomly allocated into Group A (n=37) that received allopurinol (10mg/kg B.W., per os, twice daily) for 4 consecutive months, and Group B (n=8) that were placebo-treated. Clinical signs were scored just before and at monthly intervals throughout the study period, in a blinded and independent fashion. Complete blood count, serum biochemistry profile, urinalysis, lymph node and bone marrow parasitology, IFA and enzyme-linked immunosorbent assay (ELISA) serology and bone marrow PCR were carried out at the beginning and at the end of the trial. A total of three Group A and one Group B dogs died of end stage kidney disease that developed during the trial. In Group A animals that endured the trial there was a significant improvement in the general body condition, conjunctivitis, peripheral lymphadenopathy, splenomegaly, masticatory muscle atrophy, ulcerative stomatitis, epistaxis, exfoliative dermatitis, cutaneous ulcerations, blepharitis and nasodigital hyperkeratosis. The same observation was made for anemia, lymphopenia, hyperproteinemia, hyperglobulinemia, hyperphosphatemia, increased alkaline phosphatase activity and the low albumin/globulin ratio. By contrast, no improvement of any kind was seen in Group B dogs. Lymph node and bone marrow parasite numbers were significantly decreased in Group A animals. In Group B, that occurred only in the lymph nodes. Apart from remission of clinical signs and restoration to normal of clinicopathological abnormalities, allopurinol did not eliminate Leishmania organisms, as the PCR result on bone marrow was still positive in all the dogs that finished the trial.  相似文献   

禽流感病毒抗原超滤浓缩重要参数的动态变化     

张彦杰  李爱芬 《中国兽药杂志》2007,41(10):16-17

采用世界先进的切向流超滤浓缩技术，对禽流感病毒（H9N2亚型）超滤浓缩成倍体积，测定血凝价HA和半数感染量EID50。结果HA成倍增加，废液HA呈阴性，病毒回收率达100％；受宿主鸡胚系统的影响，EID50与病毒量之间不是简单的直线关系，但病毒的活性不受影响。结果表明，所用超滤系统适用于禽流感病毒抗原的浓缩。  相似文献   

Serial analysis of serum cardiac troponin I changes and correlation with clinical findings in 46 dogs with mitral valve disease     

Zoe S. Polizopoulou  Christos K. Koutinas  Anastasia Dasopoulou  Michael Patsikas  Malcolm York  Ian Roman  Mitul Gandhi  Sonal Patel  Alexander F. Koutinas  Peter J. O'Brien 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2014,43(2):218-225

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Cutaneous involvement in canine leishmaniosis due to Leishmania infantum (syn. L. chagasi)     

Manolis N. Saridomichelakis  Alexander F. Koutinas 《Veterinary dermatology》2014,25(2):61-e22

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Food additive lactic acid production by immobilized cells of Lactobacillus brevis on delignified cellulosic material     

Elezi O  Kourkoutas Y  Koutinas AA  Kanellaki M  Bezirtzoglou E  Barnett YA  Nigam P 《Journal of agricultural and food chemistry》2003,51(18):5285-5289

Improvements in yield and productivity in lactic acid fermentation by Lactobaccilus brevis cells immobilized on delignified cellulosic (DC) material are reported. The system proved to be more efficient in comparison with the work reported by other workers. Yields of 80 and 100% conversion using glucose were obtained at 30 degrees C in 1 day of fermentation time. Lactic acid fermentation using whey as substrate was obtained at 30 degrees C in 1-1.5 days, resulting in 70% yield, whereas the remaining lactose in whey was converted to alcohol byproduct, leading to a 90% lactose exploitation and 100% conversion. Cell immobilization of L. brevis on DC material was proved by its reuses in repeated batch fermentations and through electron microscopy. A series of 10 repeated batch fermentations without any loss in cell activity showed a tendency for high operational stability. The presence of DC material resulted in a drastic drop of the fermentation time from 48 to 13 h.  相似文献   

Investigation of volatiles evolution during the alcoholic fermentation of grape must using free and immobilized cells with the help of solid phase microextraction (SPME) headspace sampling     

Mallouchos A  Komaitis M  Koutinas A  Kanellaki M 《Journal of agricultural and food chemistry》2002,50(13):3840-3848

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on delignified cellulosic material (DCM). Repeated batch fermentations were conducted using these biocatalysts and free cells, separately, at temperatures of 20, 15, and 10 degrees C. Solid phase microextraction (SPME) was used in monitoring the formation of volatile alcohols, acetate esters, and ethyl esters of fatty acids. The kinetics of volatile production were similar for free and immobilized cells. In all cases immobilized cells showed a better rate of volatile production, which was directly connected to sugar consumption. The main difference observed was in propanol production, which increased with temperature decrease for the immobilized cells, whereas it remained constant for the free ones. In the case of immobilized cells significant amounts of esters were also produced. It is well-known that esters contribute to the fruity aroma of wine. It was also established that SPME is a very sensitive, accurate, and reliable technique and can be used without any reservation in the characterization of volatile constituents of wine.  相似文献   

Cellular immunophenotyping of exfoliative dermatitis in canine leishmaniosis (Leishmania infantum)     

Papadogiannakis EI  Koutinas AF  Saridomichelakis MN  Vlemmas J  Lekkas S  Karameris A  Fytianou A 《Veterinary immunology and immunopathology》2005,104(3-4):227-237

Lymphocyte subsets, major histocompatibility complex (MHC)-II expressing cells and number of amastigotes in the epidermis and dermis were investigated immunohistochemically in 48 dogs with patent leishmaniosis, with or without exfoliative dermatitis (ED) to study the immunopathogenesis of this common cutaneous form of the disease. Skin biopsies were obtained and compared for ED sites (group A, n = 26), normal-appearing skin from the same animals (group B, n = 24), and leishmanial dogs not exhibiting ED (group C, n = 22), and normal controls (group D, n = 22). The CD3+, CD45RA+, CD4+, CD8+ (CD8a+), CD21+, and MHC-II+ cells and leishmania amastigotes were identified immunohistochemically and counted with the aid of an image analysis system. Pyogranulomatous to granulomatous dermatitis, expressed in various histopathological patterns, was noticed in all groups A and B and in half of group C dogs. In the epidermis, the low number of T-cells and their subsets did not differ significantly between groups A and B, but CD8+ outnumbered CD4+ lymphocytes in both groups. MHC-II+ expression on epidermal keratinocytes was intense in the skin with and without lesions from dogs with ED but not in group C dogs. CD3+, CD8+ and MHC-II+ cells were fewer in group C compared to group A and B dogs. In the dermis, CD3+ cells in group A animals were mainly represented by the CD8+. CD45RA+ and CD21+ cells were also seen in high numbers. MHC-II expression, potentially in lymphocytes, fibroblasts, dendritic cells, and macrophages was intense. The numbers of all cellular subpopulations in the dermis were significantly different between the groups, being highest in group A and lowest in group D. In sebaceous adenitis sites, CD4+ outnumbered CD8+ cells in contrast to the neighbouring dermis and the epidermis. The number of CD21+ and CD45RA+ cells was much lower in the inflamed sebaceous glands compared to the dermis. Finally, the number of amastigotes in the normal-appearing skin was significantly higher in the ED dogs (group B) than in those not exhibiting this cutaneous form of the disease (group C).  相似文献   

Histopathology and immunohistochemistry of canine distemper virus-induced footpad hyperkeratosis (hard Pad disease) in dogs with natural canine distemper     

Koutinas AF  Baumgärtner W  Tontis D  Polizopoulou Z  Saridomichelakis MN  Lekkas S 《Veterinary pathology》2004,41(1):2-9

Hard pad disease represents an uncommon manifestation of canine distemper virus (CDV) infection with a still uncertain pathogenesis. To study the pathogenesis of this uncommon, virally induced cutaneous lesion, the footpads of 19 dogs with naturally occurring distemper were investigated for histologic changes and distribution pattern of CDV antigen. All dogs displayed clinical signs of distemper, which had lasted from 10 to 75 days. Overt digital hyperkeratosis was observed in 12 animals (group A), whereas the footpads of the remaining seven dogs appeared normal macroscopically (group B). Orthokeratotic hyperkeratosis (12/12; 100%), irregular acanthosis (11/12; 92%), thickened rete ridges (10/12; 83%), and mild mononuclear perivascular (10/ 12; 83%) and periadnexal (7/12; 58%) dermatitis were the most common findings in dogs with hard pad disease. Surprisingly, orthokeratotic hyperkeratosis (5/7; 71%), irregular acanthosis (5/7; 71%), and thickened rete ridges (4/7; 57%) were also seen in the dogs without clinical evidence of digital hyperkeratosis. CDV-specific inclusion bodies and ballooning degeneration were not observed in the footpad epidermis of the 19 dogs. Immunohis-tochemistry revealed that CDV antigen was most frequently found in the stratum spinosum and granulosum and in the epithelial cells of the eccrine sweat glands and only rarely in the basal layer. Fibroblasts, pericytes, endothelial cells, and hair follicles were also positive in some animals. Despite the obvious difference regarding the macroscopic picture, the microscopic changes were less prominent between the animal groups. The selective infection of keratinocytes in the stratum spinosum might be the key event for the development of hard pad disease in the dog.  相似文献