Dimitra Psalla

Non-suppurative Meningoencephalitis of Unknown Origin in Cats and Dogs: an Immunohistochemical Study

Summary Non-suppurative meningoencephalitis of unknown cause is a frequent finding in dogs and cats. Fifty-three dogs and 33 cats with non-suppurative meningoencephalitis of unknown aetiology were examined immunohistochemically for 18 different infectious agents, including viruses, bacteria and prion proteinSc. In 14 (26%) of the dogs and 13 (39%) of the cats a causative agent was identified in the central nervous system (CNS), two dogs and one cat giving positive results for two infectious agents simultaneously. The study revealed infections with known causative agents (porcine herpes virus 1, feline infectious peritonitis virus, Escherichia coli) and a new disease pattern of parvovirus infection in the CNS of dogs and cats. Infection of the CNS with feline leukaemia virus was found in a cat. Five dogs and four cats gave positive results for West Nile virus (WNV) antigen. In one dog, canine parainfluenza virus antigen was detected in the brain. Four dogs and four cats gave positive results for encephalomyocarditis virus (EMCV). The significance of the detection of WNV and EMCV antigen requires further study. The aetiology remained undetermined in 39 dogs (74%) and 20 cats (61%). Although it is possible that non-infectious causes play a more important role than previously thought, infections with hitherto unrecognized agents cannot be ruled out.

Pathogenesis of experimental encephalomyocarditis: a histopathological, immunohistochemical and virological study in mice.

Mice (n=20) aged 8 weeks were infected, either by oronasal inoculation or by contact, with one of two different myocardial strains of encephalomyocarditis virus (EMCV), namely, the Greek strain 424/90 and the Belgian strain B279/95. The animals were killed at 18-59 days post-infection (dpi), except for two mice that died at 6 and 32 dpi, and samples of brain, heart, pancreas, kidney, Peyers patches, spleen, lung and thymus were processed for virological, histopathological and immunohistochemical examination. Apart from the two deaths, the experimental infection was inapparent, but virus was invariably recovered from faeces and several organs. The main histopathological lesions were focal interstitial pancreatitis, depletion of thymus and Peyers patches, and interstitial pneumonia. Additionally, in the two mice that died, multifocal interstitial myocarditis was observed. EMCV antigen was detected in the cytoplasm of pancreatic acinar cells and in macrophages of the lung and the thymus. Antigen was also detected in the cytoplasm of cardiac muscle cells from three animals, including the two that died. The results support the role of mice, in addition to rats, as reservoir hosts in the epidemiology of EMCV infections on pig farms.

Use of the laryngeal mask airway in rabbits : placement and efficacy

The laryngeal mask airway (LMA) has been used in various animal species anesthetized for the purpose of device evaluation, but the device has not been evaluated in rabbits during surgery. The authors tested the feasibility and potential advantages of using the LMA in 50 rabbits undergoing surgery under spontaneous-breathing inhalational anesthesia, focusing mainly on the technique of insertion and its efficacy. The LMA was easily inserted and no air leakage at the larynx was detected. Although four rabbits developed lingual cyanosis, this was reversible and most likely due to lingual vascular compression by the LMA. The authors conclude that the LMA is an attractive alternative to endotracheal intubation, as the mask can be inserted easily and rapidly and its correct placement is easily confirmed.

First evidence of the European wildcat (Felis silvestris silvestris) as definitive host of Angiostrongylus chabaudi

Angiostrongylus chabaudi (Strongylida, Angiostrongylidae) is a parasitic nematode described for the first time last century from the pulmonary arteries of six European wildcats (Felis silvestris silvestris) in central Italy. Since then, this parasite remained practically unknown until recently, when immature A. chabaudi have been reported from one wildcat in Germany and two domestic cats (Felis silvestris catus) in Italy. The present report describes the first record of A. chabaudi in Greece and, most importantly, the first known case of patent infection by A. chabaudi. The necropsy of a road-killed F. s. silvestris found near the lake Kerkini, in the municipality of Serres (Macedonia, Greece), revealed the presence of nematodes of both sexes in the right ventricle and the pulmonary artery of the heart. All parasites were mature adults and numerous eggs were present in the uteruses of females. The morphological characteristics of the parasites were consistent with those of A. chabaudi. Moreover, Angiostrongylus-like first stage larvae (L1) were present in the faeces of the animal that was negative for any other cardio-pulmonary parasite. Genetic examination of adult parasites and L1 confirmed the morphological identification as A. chabaudi. Histopathological examination of the lungs showed severe, multifocal to coalescing, chronic, interstitial granulomatous pneumonia due to the presence of adult parasites, larvae and eggs. These findings demonstrate for the first unequivocal time that this nematode reproduces in the European wildcat which should be ultimately considered a definitive host of A. chabaudi. Finally, the L1 of A. chabaudi are described here for the first time, opening new prospects for further studies on this neglected parasite.

Effect of locally injected autologous platelet-rich plasma on second intention wound healing of acute full-thickness skin defects in dogs

OBJECTIVES Second intention wound healing may be impaired by wound and host factors and thus more advanced therapies are required for a fast and satisfactory outcome. Platelet-rich plasma (PRP), rich in growth factors and cytokines essential for tissue repair, could improve wound healing. The purpose of this experimental study was to evaluate the effect of locally injected autologous PRP on second intention healing of acute full-thickness skin defects in dogs. METHODS Three 2 x 2 cm full-thickness skin defects were created bilaterally on the dorsolateral area of the trunk of six Beagle dogs. The wounds of one randomly selected side received PRP treatment, whereas the contralateral wounds were left untreated (controls). Wound healing was evaluated by planimetry, laser-Doppler flowmetry measurements of tissue perfusion, and histologically. RESULTS The rate of wound healing did not differ significantly between the two groups. Tissue perfusion was significantly higher in the PRP-treated group (p = 0.008) compared to controls on day 10. Histological evaluation revealed a trend towards greater collagen production and a significantly better collagen orientation (p = 0.019) in PRP-treated wounds on day 20. CLINICAL SIGNIFICANCE Locally injected autologous PRP does not accelerate the healing process, but increases tissue perfusion and may promote the formation of organized collagen bundles in acute full-thickness skin defects in dogs.

Comparison of three skin-stretching devices for closing skin defects on the limbs of dogs

Our objective was to evaluate the effectiveness of skin-stretching devices for closing defects on the extremities of dogs. Antebrachial skin defects were created on the limbs of 24 dogs randomly divided into three groups. Skin stretchers included staples and sutures passing through them (group A), sutures and hypodermic needles (group B), and Pavletic device (group C). Wounds on the left were further undermined in all groups. Tension and blood perfusion were assessed. After removing the stretchers on day 3, the defects were sutured and wound healing was clinically scored. Histological variables evaluated were cellular infiltration, edema, collagen orientation, and thickness of epidermis. Significant differences in tension were found among groups (p < 0.0005) and between measurement times for undermined (p = 0.001) or non-undermined (p < 0.0005) wounds. In contrast, blood perfusion values did not differ significantly. Clinical scores for group B seemed to be better than those for groups A and C, but differences were not significant. Primary wound closure using the Pavletic device was not feasible. No significant differences in histological variables were found between groups. Skin stretching with staples or hypodermic needles resulted in successful wound management with minor side effects on skin histology and circulation.

Effects of single dose administration of ifosfamide on testes and semen characteristics in the rabbit.

The effects on testes and semen characteristics were studied in the rabbit after a single dose administration of the chemotherapeutic agent ifosfamide. Sexually mature male rabbits received a single intravenous injection of either 0, 60, 90, 120, or 240mg/kg body weight ifosfamide. Two phases of experimentation were conducted, lasting 1 and 18 weeks, respectively, at the end of which half of the animals from each treatment group were euthanized. Reproductive organs weighing, as well as testicular qualitative and quantitative histological examinations were performed 1 and 18 weeks post-treatment, while semen quality and libido were evaluated on a weekly basis. A decrease in the paired testes weight (90, 120, and 240mg/kg groups) and the accessory sex glands weight (240mg/kg group) were noted 1 week post-treatment. Although no histopathologic lesions or significant changes in the quantitative histologic examination were observed, semen quality examination revealed transient oligospermia (60, 90, 120, and 240mg/kg groups), teratozoospermia (120 and 240mg/kg groups), and asthenozoospermia (240mg/kg group), which returned to normal by the 6th (60 and 90mg/kg groups), 7th (120mg/kg group), or 8th week after treatment (240mg/kg group). Libido remained normal. The results suggest that ifosfamide, at a single dose, causes transient and dose-dependent depression of spermatocytogenesis (240mg/kg), spermiogenesis (60, 90, and 120mg/kg), and sperm maturation in the epididymis (240mg/kg).

Locally injected autologous platelet-rich plasma enhanced tissue perfusion and improved survival of long subdermal plexus skin flaps in dogs

OBJECTIVES Distal flap necrosis remains a major complication in subdermal plexus (random) skin flaps. Platelet-rich plasma (PRP) has been shown to improve the survival of ischemic random skin flaps in rats. The objective of this study was to evaluate the effect of locally injected autologous PRP on the survival of long (5:1 length-to-width ratio) subdermal plexus skin flaps in dogs. METHODS A 2x10 cm subdermal plexus skin flap was created bilaterally on the abdominal wall of six Beagle dogs. One randomly selected side received 2.5 ml of fresh auto-logous PRP injected evenly between sutures underneath the flap, whereas the other side was left untreated (control). Skin flap survival was evaluated macroscopically, histologically and by laser-Doppler flowmetry measurements of tissue perfusion. RESULTS Flap percentage survival on day 10 (96.3% versus 74.5%; p = 0.046) and tissue perfusion (p <0.036) were significantly higher in PRP-treated flaps compared with controls. Histologically, there was less oedema in PRP-treated flaps compared to controls (p = 0.01), whereas collagen production and angiogenesis did not differ significantly between the two groups. CLINICAL SIGNIFICANCE The use of locally injected autologous PRP increases tissue perfusion and improves the survival of long subdermal plexus skin flaps in dogs.

Immunohistochemical expression of dogTERT in canine testicular tumours in relation to PCNA, ki67 and p53 expression.

The objective of the study was to determine the immunohistochemical expression of canine TERT in canine testicular tumours comparing two different antibodies for TERT, and to correlate them with well established markers specific to dividing cells such as PCNA and ki67, and with expression of the p53 tumour suppressor gene. The study included 36 cases of canine testicular tumours, which were categorized as 12 Sertoli Cell Tumours (SCT), 20 seminomas, 3 interstitial cell tumours and 1 mixed germ cell-sex cord stromal tumour (MT). Two antibodies for hTERT were examined; a highly specific TERT antibody, RCK-hTERT, was evaluated for the first time. Immunodetection of RCK-hTERT was observed in 31% of tumours examined (6/20 Seminomas, 4/12 SCT, 1/3 interstitial cell tumour and 0/1 mixed germ cell-sex cord stromal tumour), while the NCL-hTERT in 67% of them (15/20 Seminomas, 6/12 SCT, 3/3 interstitial cell tumour and 0/1 ΜΤ). PCNA immunoreactivity was detected in all cases. Regarding ki67, 3 SCT, 12 seminomas and all interstitial cell tumours showed clear immunoreaction. p53 immunoreactivity was detected in 6 SCT, 15 seminomas and all interstitial cell tumours. The immunohistochemical expression of both TERT antibodies are discussed and compared in order to clarify their potential usefulness in canine testicular malignancies in relation to the expression of well known cell cycle markers. Our results indicate that TERT and PCNA are useful proliferation markers but not helpful to evaluate prognosis. Instead of that ki67 and p53 could be used for predicting aggressiveness in this group of tumours

Nodular granulomatous glossitis as the sole clinical sign in canine leishmaniosis

A 5.5-year-old, intact male Rottweiler dog was admitted with a history of multifocal nodular tongue lesions which progressively deteriorated during the previous year. Physical examination revealed several reddish nodules with central depression on the surface of the tongue in an otherwise healthy dog. Clinicopathologic abnormalities included eosinophilia and hyperproteinemia. Lingual nodule cytopathology, histopathology, and immunohistochemistry revealed Leishmania spp. amastigotes and a severe granulomatous glossitis. The dog was also seroreactive to L infantum antigens by an indirect immunofluorescence assay. Clinical reevaluation 3 months after the institution of treatment with allopurinol and miltefosine indicated that the nodular lesions had completely regressed. In endemic areas, lingual nodular lesions may rarely be the sole clinical sign of canine leishmaniosis. Standard medical treatment may provide an excellent prognosis.