Adrian Gal

Multidisciplinary analysis of Knemidocoptes jamaicensis parasitising the Common Chaffinch, Fringilla coelebs: proofs for a multispecies complex?

The number of studies discussing the pathology and host specificity in Knemidocoptinae is very limited. In Knemidocoptes jamaicensis, the host specificity seems to be very broad, and there is a clear morphological variability in individuals originating from various bird species; hence, serious doubts appear about the species status of this mite. We report a multidisciplinary approach to the taxonomy, morphology, ecology, and pathology of K. jamaicensis. The source of the mites in our study was a second year aged female of the Common Chaffinch, Fringilla coelebs, which accidentally died in the mist net during a field study in Dumbrava, Cluj County, Romania in March 2011. Comparisons of the biometrical data regarding the body dimensions, length of certain setae, and distances between bases of dorsal setae with other published data showed a great variability of certain measurements between populations infecting various hosts and localities and sometimes even within single populations. Gross and histologic lesions consisted in severe bilateral orthokeratotic hyperkeratosis and epidermal spongiosis. Lesions also involved the skin of the joints. Skin inflammation was absent, and no lesions were noticed in the metatarsus bone. Following molecular analysis, the 518-base-long sequence differed from the published 18S rDNA in nine positions. Additionally, our paper reports for the first time the DNA barcode sequences of K. jamaicensis and, together with the synoptic analysis of host spectrum, geographical distribution and morphological variability it brings important evidences to sustain the hypothesis of multispecies complex for K. jamaicensis.

Effects of astaxanthin supplementation on chemically induced tumorigenesis in Wistar rats

BackgroundAstaxanthin (ASTA) is a fat-soluble xanthophyll with powerful antioxidant functions. It is extracted from e.g. salmon, an important food source for certain human populations known to have a reduced risk of tumor development. It is possible that ASTA plays a role in cancer chemoprevention in such populations. The purpose of this study was to investigate the effects of dietary ASTA on chemically induced mammary tumorigenesis using N-methyl-N-nitroso-urea (MNU) in immature Wistar rats.MethodsThirty-six 37 days old juvenile female Wistar rats were at random allocated to 4 groups of which Groups 1 and 2 received a single dose of 55 mg MNU/kg body weight. The effects of ASTA was evaluated by giving rats of Groups 2 and 4 a dose of 50 mg ASTA/kg/day for the entire duration of the study. Group 3 rats received feed added alimentary oil.Necropsy and histopathological examinations were carried out on each rat 14 months after the administration of MNU. Haematological values and antioxidative status were determined. Oxidative stress was evaluated by monitoring superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in hepatic tissue. Lipid peroxidation and carbonylation of proteins was determined in protein extracts from the liver.ResultsTumor development occurred only in rats of Groups 1 and 2, i.e. MNU exposed animals. Frequency of tumor development in general and average number of tumors per animal were insignificant between these two groups. Mammary gland tumors developed in equal frequencies in Group 1 and 2 rats, respectively. Although only rather few tumors were found in the mammary glands, a substantial number of other tumors were found in Group 1 and 2 rats, but at equal rates.Biochemical analyses showed significant higher levels of GPx, malondialdehyde and dinitrophenylhydrazine in Group 1 rats that for rats in all other groups thus indicating protective effects of ASTA on MNU induced hepatic oxidative stress.ConclusionsSupplementation with ASTA did not reduce tumorigenesis induced by MNU in Wistar rats. However, supplementation with ASTA seemed to have anti-inflammatory effects.

Vascular abnormalities of the distal deep digital flexor tendon in 8 draught horses identified on histological examination

The purpose of this study was to provide a detailed description of the vascular changes in the distal part of deep digital flexor tendon (DDFT). Eight isolated forelimbs were collected from 8 horses with DDF tendinopathy diagnosed post-mortem by ultrasound and gross anatomopathological examination. The samples were fixed in 10% neutral buffered formalin, softened in 4% phenol and dehydrated with ethylic alcohol. Goldners Trichrome staining method was used. The histopathological examination revealed vascular proliferation associated with structural disorders of blood vessels. Angiogenesis, fibroplasia and consecutive hypertrophy of the vascular wall with or without vascular occlusion were the most common findings. Other histopathological findings were: endothelial cell edema, progressive metaplasia from squamous to cubic cells, vascular wall hyalinization, endothelial cells apoptosis/necrosis and endothelial desquamation. These results demonstrated damage of the distal deep digital flexor tendon vasculature which may progressively alter the structural integrity of the tendon and contribute to degenerative lesions.

Metastatic cholangiocarcinoma in a llama (Lama glama)

A 2-year-old female llama (Lama glama), from a private zoological park, with anorexia, ataxia, dyspnea, ascites, and emaciation, was necropsied. Gross inspection, and cytological, histological, and immunohistochemical analyses were performed. A firm, white, multinodular mass, 25 cm in diameter was found in the liver parenchyma. Similar nodules up to 3 cm were found in lymph nodes, lung, diaphragm, and peritoneum. Histologically, the affected organs were replaced by an infiltrative growth of undifferentiated neoplastic cells. Immunohistochemically, neoplastic cells were positive for pancytokeratin (panCK), CK20, and CK19, and negative for thyroid transcription factor 1, CK7, and carcinoembryonic antigen. A diagnosis of poorly differentiated metastatic cholangiocarcinoma was made.

Lethal herpesvirosis in 16 captive horned vipers (Vipera ammodytes ammodytes): pathological and ultrastructural findings

Sixteen captive adult horned vipers (Vipera ammodytes ammodytes) were submitted for necropsy examination following a 2-week history of lethargy, anorexia and dyspnoea. Gross lesions included widespread haemorrhage, serosanguineous effusions in the body cavities and multiple pinpoint white to yellow foci in the liver. Microscopically, there was multifocal hepatic coagulative necrosis associated with intranuclear acidophilic inclusion bodies in sinusoidal endothelial cells. Similar endothelial lesions were observed in the myocardium, fat bodies, kidneys and spleen. Transmission electron microscopy revealed numerous virions (100-110 nm) in the nuclei of endothelial cells and intracytoplasmic enveloped virions (140-150 nm) were also found. The gross and histological findings and the ultrastructural features of the intranuclear inclusions and viral particles were consistent with herpesviral infection. To the best of our knowledge, these are the first reported cases of a lethal herpesvirosis in horned vipers and the second report in snakes.

Mesothelial metaplasia in European pond turtle, Emys orbicularis (Testudines: Emydidae) infected with Spiroxys contortus (Nematoda: Spirurida)

SummaryThe mesothelium consists of a monolayer of specialized epithelial cells that line the surfaces of the peritoneal, pleural, and pericardial cavities. The present study reports mesothelial changes associated with larval stages of the spirurid nematode Spiroxys contortus in a naturally infected European pond turtle. Between 2002 and 2007, eight European pond turtle, Emys orbicularis were brought for necropsy. In one adult male turtle, necropsy revealed the presence of gastric and mesothelial changes associated with gastric infestation with the nematode Spiroxys contortus. Gastrointestinal samples with gross lesions were collected and processed by paraffin technique for further histological examination. Mesothelial cuboidal to columnar or pseudostratified focal progressive metaplasia was observed in visceral coelomic cavity. Intracytoplasmic acidophilic hyaline inclusions were also observed in metaplastic mesothelial cells, most of them being placed to the apical cell’s poles. The significance of our findings is discussed in the light of current knowledge regarding the mesothelial cells.

New Aspects of Graft-induced Central Plasticity and Neuroregeneration at T 10 in a Rat ModelâÂÂ

Objective: Retest Brunelli’s graft induced glutamate neurotransmitter switch at the neuromuscular junction in rat for the translation of new aspects of central plasticity concepts for human reconstructive surgery in spinal cord lesions. Methods: Randomized double blind controlled study in rat, which was limited to 30 animals (Charles River, 220 to 280 g). Ethical research approval was obtained from the Animal Research Committee of the University Hospital Schleswig Holstein, UK-SH, Lubeck, D, and legitimated by the Governmental Department of Agriculture, Natural Environments and Agriculture Kiel, D, in compliance with the European Commission Recommendation to retest and review of graft-induced glutamatergic regeneration and /or cholinergic co-transmission at the neuromuscular junction for reinnervation of the skeletal obliquus abdominis internus muscle fibers. Assessments were performed to demonstrate pharmacological neuromodulation after attaching the lateral corticospinal tract at T10 to the bisected skeletal motor nerve. Medication was administered for 14 days postoperatively–a) verum Cerebrolysin® IP=12–b) shams NaCl 0.9% IP=11, and c) 7 controls (nil). 2nd Op at day 90 (16 surviving rats) for open proof of reinnervation and its type by a) CMAP, b)Vecuronium® application. Fast Blue® labeling were performed. 10 days later, on the 3rd Op N=15, euthanasia and organ fixation were performed. Extensive histology-morphology examinations were performed in Cluj. Results: Eight rats showed positive CMAPs. Reinnervation and neuromodulation were demonstrated by counting and comparison of the grafted muscle fibers diameter. Four CAMP- positive- rats received Vecuronium®: 1 CERE and 1 NaCl each demonstrated a loss of amplitude respectively two an incomplete muscle blockage due to the coexistence of glutamatergic and cholinergic neurotransmission. Confirmation of the VGluT2 in axons was observed by immunofluorescence. FB+ neurons were observed in many Rexed laminae in grafted spinal cord, but not in the brain. Conclusion: The coexistence of graft-induced cholinergic and glutamatergic neurotransmission and a great capacity of lower motor neurons and other types of spinal neurons to regenerate were observed. Because of limited animals, pharmacological neuromodulation requires further investigation.

Description of a Feline Injection-Site Fibrosarcoma with Metastasis in the Cerebellum

Abstract Feline injection-site sarcomas were initially described by Hendrick and Goldschmidt (1991). The link between vaccination site and sarcoma occurrence suggested the term of vaccine-associated sarcomas. Our paper describes an unusual feline injection-site fibrosarcoma with cerebellar metastasis. A 7-year-old female domestic short-hair cat was submitted to the Pathology Department (Faculty of Veterinary Medicine Cluj- Napoca, Romania) for necropsy. A subcutaneous tumor (relapse) in the interscapular region and multiple metastatic masses in the lungs, kidneys, subcutaneous tissue (scapular and thigh regions) and cerebellum were observed. Cytological, histological and immunohistochemical (for vimentin, desmin, multi-cytokeratin, α-smooth muscle actin, S100 and CD45) analyses from all tumors were performed. Cytological examination identifi ed highly pleomorphic spindle-shaped cells admixed with neoplastic multinucleated giant cells. Histologically, all neoplastic masses were composed of numerous spindle cells arranged into interlacing bundles. Extensive intratumoral areas of necrosis along with a neutrophilic infiltrate were also detected. A fibrillary material was present among neoplastic cells (green stained by Masson’s trichrome method), suggesting a collagenous structure. In all tumors assessed, immunohistochemistry showed an intense reaction only for vimentin in numerous neoplastic cells. Based on the history, gross, cytological, histological and immunohistochemical data, the final diagnosis was recurrent feline injection-site fibrosarcoma, with multiple metastases (including in the cerebellum). Overall, an uncommon case of feline injection-site fibrosarcoma has been reported. This is the first consistent record of an injection site sarcoma in a cat with cerebellar metastasis.

Comparative immunohistochemical assessment of the effect of repetitive anesthesia with isoflurane and sevoflurane on rat liver

BACKGROUND AND OBJECTIVES Inhalation anesthetics are used in human, as well as veterinary medical practice. In the present study we investigated the effect of isoflurane and sevoflurane on rat hepatocytes. METHODS A total of 40 Wistar female rats were used in this study. Animals were divided in groups of 5 rats. Groups IM, SM served as control groups. Groups I1, I2, I3 were used to study isoflurane and S1, S2, S3 for sevoflurane study. They were anesthetized 3 times, for 2h long, at 2 days interval with a concentration of: 1.5% isoflurane (I1, I2, I3) and 2% sevoflurane (S1, S2, S3). The oxygen supply throughout the anesthesia was 1LO2/min. Groups IM, IS, I1, S1 were sacrificed immediately after the last anesthesia. Groups I2, S2 were sacrificed 6h after the last anesthesia, and groups I3, S3, 24h post-anesthesia. Liver samples were harvested to highlight caspase-3 in apoptotic hepatocytes. RESULTS Following isoflurane administration, there were less than 1% cells in apoptosis highlighted in rat livers from groups IM, I1 and I2. At 24h post-anesthesia (group I3), a small number of apoptotic hepatocytes was highlighted (around 3.23% cells in apoptosis), with a strictly periacinar disposition, randomly distributed in a small number of hepatic lobules. After sevoflurane administration, less than 1% apoptotic hepatocytes were identified at all control moments throughout the study. CONCLUSIONS The results suggest that the anesthetics do not present a considerable hepatotoxicity. The comparative assessment of the two anesthetics shows that sevoflurane is superior to isoflurane.

Histological evidence concerning the osseointegration of titanium implants in the fractured rabbit femur

Stabilization of the broken bone is achieved using biocompatible materials. Since histology is still considered the gold standard technique for the assessment of bone formation around metallic implants, this report investigated the titanium implant integration in the accidentally broken bone in rabbits. The experimental protocol was reviewed and approved by the Ethical Committee of the Faculty of Medicine and Pharmacy Oradea, Romania. Holes were drilled in the diaphysis of the femur, and titanium implants were inserted in the created bone defect. In two subjects, fractures occurred on days two and three after the metallic alloy implantation. The other two rabbits presented no fractures following the surgical procedure. The rabbits were euthanized and the bones (with metallic implants) were harvested for histopathological investigation. Following decalcification, the bone samples were processed using the standard paraffin technique and stained by Goldner’s trichrome procedure. In subjects with a perfect immobilization of the titanium implants, the osseointegration occurred with minimal callus formation (i.e. primary cortical healing). In rabbits with bone fractures, the callus was more exuberant. A progressive replacement of the granulation tissue with hyaline cartilage and woven bone occurred soon after. The former aspects suggested an indirect metaplasia in the created callus. In all subjects, no inflammatory cells were identified in the created callus. The bone regeneration occurred either by primary cortical healing (in perfectly immobilized titanium implants) or by a process similar to the endochondral ossification (in poorly immobilized titanium implants following accidental post-implantation bones fracture).