Vesna Lalošević

High infection rate of zoonotic Eucoleus aerophilus infection in foxes from Serbia

The respiratory capillariid nematode Eucoleus aerophilus (Creplin, 1839) infects wild and domestic carnivores and, occasionally, humans. Thus far, a dozen of human infections have been published in the literature but it cannot be ruled out that lung capillariosis is underdiagnosed in human medicine. Also, the apparent spreading of E. aerophilus in different geographic areas spurs new studies on the epidemiology of this nematode. After the recognition of the first human case of E. aerophilus infection in Serbia, there is a significant merit in enhancing knowledge on the distribution of the nematode. In the present work the infection rate of pulmonary capillariosis was investigated in 70 red foxes (Vulpes vulpes) from the northern part of Serbia by autopsy. The estimated infection rate with Eucoleus aerophilus was 84%. In contrast, by copromicroscopic examination only 38% of foxes were positive. In addition, 10 foxes were investigated for the closely related species in nasal cavity, Eucoleus boehmi, and nine were positive. Our study demonstrates one of the highest infection rates of pulmonary capillariosis in foxes over the world.

Neospora caninum antibodies in dairy cows and domestic dogs from Vojvodina, Serbia

Neospora caninum, the causative agent of neosporosis, is a protozoan parasite responsible for high rate of abortion in cattle worldwide. In dogs, consequences of infection vary from severe neuromuscular disorders to asymptomatic infection and shedding of environmentally resistant oocysts. In this study, we determined the occurrence of N. caninum antibodies in dairy cattle and dogs in Vojvodina (Northern Province of Serbia) and possible risk factors. N. caninum antibodies were found in 15.4% (55/356, CI 95%:12.0–19.6) of cows and 17.2% (17/99, CI 95%: 10.8–26.2) of dogs. Cows from smallholdings showed significantly greater odds (OR = 5.28, CI 95%: 2.0–13.6, p = 0.0006) of being seropositive in comparison to the farm cows. Epidemiological importance of results is discussed.

Borrelia-like organism in heart capillaries of patient with Lyme-disease seen by electron microscopy.

A case of a patient who developed an acute myocarditis due to Lyme disease is reported. An increased serum antibody titer to Borrelia burgdorferi suggested a diagnosis and in addition of basic clinical methods, endomyocardial biopsy performed and analyzed by transmission electron microscopy. The lumen of myocardial capillaries was founded mostly filled with detritus and fibrin precipitate, between them several bacterial fragments were identified. The electron-microscopic characteristics of the microorganisms in this specimen, revealing irregularly coiled appearance and consistent thickness of 0.2 μm, correspond to the spiral-like structure of Lyme disease borrelia. The presence of fibrin deposits on the capillary endothelium and necrosis of myocardiocytes, suggests that the cardiopathy in our patient was represent borrelia-mediated damage of the heart microcirculation.

Current differential diagnosis of hypereosinophilic syndrome

INTRODUCTION Hypereosinophilic syndrome (HES) is a group of idiopathic disorders associated with single or multiple organ system dysfunction. HES must be distinguished from reactive eosinophilia in parasitic infections, allergic diseases, and especially from hematological diseases of clonal origin. REACTIVE EOSINOPHILIA DUE TO INFECTIOUS AND PARASITIC DISEASES Tissue helminth infections, especially toxocariasis, cause severe and long-standing hypereosinophilia. Despite specific therapy, eosinophilia may persist for over a year after diagnosis, and decreases slowly. Other helminth infections, such as trichinellosis, strongyloidosis, and rarely taeniasis and cysticercosis may also be diagnosed in patients with eosinophilia. HEMATOLOGIC AND OTHER NEOPLASTIC DISEASES Numerous neoplastic diseases, like Hodgkins and other malignant lymphomas, myeloproliferative diseases, systemic mastocytosis etc., may be associated with marked eosinophilia. We had two patients with clinical and histological features resembling chronic eosinophilic leukemia, and many others with T-cell lymphoma, planocellular or adenocarcinoma etc. where eosinophilia persisted DRUG-INDUCED EOSINOPHILIA: Drugs associated with eosinophilia include penicillins, tetracyclines, especially minocycline, clarithromycin, tetrazepam, mefloquine, and many, others. Toxins associated with L-tryptophan cause eosinophilia-myalgia syndrome and toxic oil syndrome, also belonging in this group. Treatment includes drug discontinuation and administration of corticosteroids. HYPEREOSINOPHILIA WITH ORGAN DYSFUNCTION Many severe diseases, such as sarcoidosis, Churg-Strauss syndrome, pemphigus vulgaris, eosinophilic gastrointestinal diseases, inflammatory bowel disease and many others are associated with hypereosinophilia and target organ damage, e.g. involvement of the heart, lungs, skin, or nervous tissue. CONCLUSION Eosinophilia can be classified as either familial or acquired. Hypereosinophilic syndrome is a subcategory of idiopathic eosinophilia. If the differential diagnosis of hypereosinophilia fails to resolve the etiology succesfully, the diagnosis of idiopathic HES remains.

Detection of Viable Toxoplasma gondii in Free-Range Pigs from the Special Nature Reserve of Zasavica

Summary Toxoplasma gondii is considered one of the most successful parasites of humans and animals. The ingestion of viable cysts through the consumption of undercooked pork is recognized as a significant route of human infection with T. gondii. The aim of this study was to investigate the presence of viable parasite in tissues of free-range pigs from the Zasavica Special Nature Reserve. All pigs were of the Mangulica breed, raised in a traditional way. The serological screening was performed using a modified agglutination test (MAT). The isolation of viable T. gondii was attempted by a bioassay of pig heart tissue in mice, while the real-time polymerase chain reaction (qPCR) targeted at the 529 bp repetitive element of T. gondii was used to detect parasitic DNA in digested hearts. Specific antibodies were detected in 12 out of 18 pigs examined. The bioassay was performed for five MAT-positive and one MAT-negative pig, and a total of three isolates were obtained. qPCR was performed for all samples, including one MAT-negative sample that was not bioassayed. The presence of T. gondii DNA was confirmed in all hearts with a positive bioassay as well as in one originating from seropositive and one from seronegative pig whose hearts were not bioassayed. The successful isolation of viable cysts, presence of risk factors (such as older age at the time of slaughter) and increased contact with the environment, along with the great appreciation of Serbian consumers towards home-cured Mangulica’s meat, make this breed worthy of consideration as a potentially important reservoir of human infection.

Disodium Cromolyn and Anti-podoplanin Antibodies Strongly Inhibit Growth of BHK 21/C13-derived Fibrosarcoma in a Chick Embryo Chorioallantoic Membrane Model

Aim: To characterize baby hamster kidney fibroblast (BHK 21/C13) cells and test the effects of antibodies against podoplanin and disodium cromolyn on BHK 21/C13 cell line-derived tumors grown on chick embryo chorioallantoic membrane (CAM). Material and Methods: BHK 21/C13 cell-derived fibrosarcomas developed in hamsters were implanted on CAM and treated with anti-podoplanin antibodies and disodium cromolyn. BHK 21/C13 cell immunophenotype was assessed. Results: Fibrosarcoma cells were positive for vimentin, CD117, smooth muscle actin, vascular endothelial growth factor epidermal growth factor receptor, homebox prospero gene 1 and negative for platelet-derived growth factor B, neuron-specific enolase, S100, CD34, Ewing sarcoma and podoplanin. CAM-grown fibrosarcomas were highly sensitive to disodium cromolyn and anti-podoplanin antibodies. Conclusion: Immunophenotyping BHK 21/C13 cells and their response to drugs represent the first step in revealing cell line utility and a reliable tool for experimental cancer research.

Correlation between Different Helicobacter Morphotypes and Histological Changes in Pig Gastric Mucosa

Background: Two distinct morphologic types of bacteria which belong to the Helicobacter species, have been described in pigs: once or twice curved Helicobacter pylori-like bacteria (HLO) and the multicoiled, Gastrospirillum-like bacteria (GLO). The objective of this study was to determine the incidence of Helicobacter spp. using modified Giemsa stain and to define the relationship between presence of Helicobacter spp. and histopathological changes of gastric mucosa in pigs. Materials, Methods & Results: A total of 120 pig stomachs (60 from intensive and 60 from extensive breeding) were enrolled in this study and 240 fragments of fundic and pyloric mucosa were taken for histopathological examination. By modified Giemsa staining of gastric mucosa, Helicobacter-like organisms were confirmed in 4/60 (6.67%) of pigs in intensive and 5/60 (8.33%) of pigs in extensive breeding. The incidence of tightly spiral shaped Gastrospirillum-like organisms in pigs of intensive and extensive breeding were 5/60 (8.33%) and 9/60 (15%), respectively. The severity of gastritis was scored to the Sydney System with some modifications. There was no significant difference between HLO-positive and HLO-negative fundic mucosa in pigs of both breeding systems. In contrary, there was significant difference between HLO-positive and HLO-negative pyloric mucosa of pigs in intensive (P < 0.001) and in extensive breeding (P < 0.05). In intensive breeding, there was significant difference between GLO-positive and GLO-negative fundic mucosa (P < 0.05), while there was no significant difference between GLO-positive and GLO-negative pyloric mucosa. Discussion: The prevalence of Helicobacter spp. bacteria were in coherence with literature data. The higher prevalence of both morfological type of bacteria, were considered in pigs in extensive breeding. the hygienic conditions and managment factors in pigs farm are the possible impact for higher bacterial transmission. The association of high prevalence of H. pylori and poor hygienic condition was shown by epidemiological studies conducted on humans. In both, humans and pigs, the presence of H. pylori correlates with an inflammatory response, but there are differences in inflammatory cell population. In H. pylori infected humans, neutrophils composed the bulk of cellular infiltrate, while in pigs, the primary inflammatory cell was the lymphocyte, which is in accordance to results published by others autors. Thus it indicates that different hosts exhibit a different pathohistological response to the Helicobacter spp. infection. In human as well as in veterinary pathology, the fact of the different pathogenicity of various Helicobacter species is well known. In all HLO-positive pyloric mucosa, moderate to severe focal or diffuse infiltration of mononuclear cells and lymphoid follicles with germinal centers, were observed. A similar conclusion was drawn from results of an experimental infection study in pigs. There was signifficance between HLO-positive and HLO-negative pyloric mucosa in both, intensive and extensive breeding. In the contrast, GLO were not associated with the presence of severe gastritis, but only with mild to moderate superficial infiltration of lymphocytes and plasma cells in both, fundic and pyloric mucosa. There was no significant difference between GLO-positive and GLO-negative pyloric mucosa of pigs in both breeding systems. Despite the low gastritis score of fundic mucosa in pigs in intensive breeding, there was a significance difference between in GLO-positive and GLO-negative fundic mucosa. It is believed that the possible reason of such results is the meals with low fiber content and low particles size. These results suggest that the presence of HLO, but not of GLO is associated with the pyloric gastritis in pigs.

Nematophagous Activity of Duddingtonia Flagrans MUCL 9827 Against Sheep Gastrointestinal Nematodes

Summary Gastro-intestinal nematodes (GIN) of sheep are one of major constraints in grazing production systems worldwide. Control is commonly achieved using anthelmintics, but global occurrence of anthelmintic resistance to different drugs and the emergence of multi-resistant GIN species seriously limit the efficiency of their use. Therefore, integrated parasite management is widely recommended, with nematophagous fungi as one of control tools. Duddingtonia flagrans is one of the most used species, with various effect of different isolates. In previously performed coproculture assay, we showed low efficacy of D. flagrans MUCL 9827 against infective larvae (L3) of sheep GIN. The aim of current experiment was to reevaluate its nematophagous potential, using the medium where direct interaction between the fungus and L3 could be observed. Nematophagous activity was tested on 2% water agar with addition of chloramphenicol on three series of plates seeded with 500 and 1000 chlamidospores and agar blocks with 7 days old mycelium. At Days 0 and 5, 500 L3 of sheep GIN were added to test the trapping activity. The cultures, including control plates with only L3, were incubated at 25°C for 10 days, followed by evaluation of their number and reduction percentage. Nematophagous activity of D. flagrans MUCL 9827 against L3 was clearly demonstrated. However, the overall efficacy was poor since trapping was observed only in one out of nine plates containing fungal material. Potential reasons for such poor performance of the isolate of fungal species, otherwise known as successful in trapping animal parasitic nematodes, are discussed.