RAMONA STOICESCU*, VIVIANA CUZIC*, ADRIANA BĂLAŞA*, ELENA GORUN**
*Universitatea „Ovidius”; **Laboratorul Prodiagnostic, Constanţa

Pathological changes in digestive tract can be induced by bacterial, viral, parasitic, fungal infections and malignancies (with high frequency in HIVinfected-AIDS in children). Oral manifestations are represented by candidiasis, pigmented nodules and ulcers. Esophageal manifestations reflect Candida albicans infection, cytomegalovirus or herpes virus infections. Diarrhea is a common cause of morbidity in children and is the leading cause of death in the first year of life in developing countries. Persistent diarrhea is correlated with a previous significant immune suppression and impaired growth and increases the risk of death in HIV positive children. Common microorganisms causing diarrhea in children with HIV infection include: rotaviruses, Shigella, Salmonella, Campylobacter and Escherichia coli. Other opportunistic infections are produced by Cryptosporidium, Isosporidium, Cytomegalovirus and atypical mycobacteria. Material and methods. The study was retrospectively conducted by documentary analysis method in the Department of Pediatrics Constanţa County Hospital for a period of 13 years (1.01.1990 – 31.12.2002), on a group of 153 children diagnosed with HIV/AIDS. The patients were followed up in dynamics up to December 31, 2008. Digestive impairment was seen in all patients in the group of study. Results. In the group of 153 HIV positive patients, candidiasis was the most common oral lesion (present in 39.2% cases). It was followed by intestinal infections manifested by diarrhea that dominated the clinical picture of HIV infected patients. Out of 138 cases of HIV infected children who had infectious diarrhoea, opportunistic parasitic infections were present in 105 cases. Parasitological examination revealed the presence of poliparasitism (73.33%) compared with those infected with only one species of parasites. The highest number of cases were caused by Protozoa. Conclusions. Digestive manifestations are common in HIV infected children. The clinical picture depends on the causative agent and the host’s immune status. Enteritis caused by some opportunistic infections in patients with significant immunosuppression may lead to severe dehydration and a significant weight loss, causing a severe form of chronic diarrhoea.

D. STERIU, MARIANA POP, IONELA NEAGOE
INCDMI ”Cantacuzino” Bucureşti

Toxoplasmosis is a common oportunistic parasitic infection produced by Toxoplasma gondii sporozoan. Toxoplasmosis may be acquired by ingestion of oocysts or by ingestion of tissue inhabiting stages of the parasite. The domestic cat is probably the major source of contamination with oocysts, but infection in humans is probably most often the result of ingestion of tissue cysts contained in raw or undercooked meat. Infection may be congenitally or postnatally acquired. Postnatally acquired infection may be localized or generalized. Lymphadenitis is the most frequently observed clinical form of toxoplamosis in humans. Congenital infection occurs only when a woman becomes infected during pregnancy and severity of the disease in the fetus may depend on the stage of pregnancy when the mother becomes infected. Program for prenatal screening and diagnosis in the pregnant woman is presented.

CRISTINA ELENA STAICU
DSVSA Constanţa

Maedi Visna is a viral disease of sheep and goats, caused by a lentivirus sheep and characterized by chronic inflammation of the lung, mammary gland, joints and central nervous system. In the present study was intended presence of parasites in sheep infected with Maedi Visna. For this research samples were analyzed organs (lung, liver and CNS) from 14 cadavers and sheep for parasitological examination were tested coproparazitologic 25 faecal samples from sheep aged 1 year and 10 years.

S. SIKÓ BARABÁSI1, P. DEPLAZES2, V. COZMA3, C. CEICA4, E. AVRAM4,
I. BOGOLIN5, SIMONA POPESCU6, RALUCA POPESCU7
1Asociaţia ECHINO-NEWS România; 2Institutul de Parazitologie, Universitatea din Zürich, Elveţia;
3FMV USAMV Cluj-Napoca; 4DSVSA Satu Mare; 5DSVSA Bistriţa Năsăud; 6LSVSA Bihor; 7LSVSA Covasna

The occurrence of zoonotic tapworm, Echinococcus multilocularis, in definitive hosts, in intermediate or aberrant hostes has been described in many east-European countries. So fare, the adult stage of the parasite wasn’t described in red foxes in Romania. Therefore, between August 2007 – August 2009, we are investigated 464 european red foxes (Vulpes vulpes L.) for intestinal helminthes. The investigated foxes small intestines samples in 12.28% were infected with trematodes, 20.47% with cestodes and 15.98% in nematodes. In 8 (1,71%) of the samples, from the north-west region of Romania, we are detected Echinococcus multilocularis (between 8 and > 100 intestinal stages per sample). The morphologically identification was confirmed by a multiplex PCR, followed by sequence analysis. The Echinococcus-specific antigens detection by copro-ELISA test (CHEKIT-Echinotest biphasic enzyme immunoassay kit), in 100 fecal samples was positive in 13%. This is the first confirmation for the presence of Echinococcus multilocularis in the Romanian fox population.

- CASE REPORT -

DOINA PĂTRAŞCU, GEORGIANA CĂCIULĂ
DSVSA Caraş-Severin

Etiologic agents of animals mange and infestations can occur in humans when a closer cohabitation. In June 2008, a 9 week old Bichon Maltese dog was brought to our laboratory with severe and generalized pruritus and restlessness. The owner of the dog was found to be infested with Sarcoptes scabiei by a specialist in human dermatology so that the dog was sent to our laboratory with presumptive diagnosis of sarcoptic mange. The owner of the dog, her son aged 9 years and two relatives of them, had itching, erythematosis and papuloses lesions on the shoulder blades, coastal and abdomen areas. The animal was purchased three weeks before the development of the lesions on the owner from a pet shop from Timişoara. At purchase the dog had a valid health certificate issued by a veterinarian that stated the animal was healthy. The skin lesions, congestion and hemorrhage, skin trauma due to intense itching localized on the abdomen, armpits, groin and the ear internal conches in conjunction with the age of the animal suggested the animal was suffering from sarcoptic mange. On the examination of the skin scrapings S. scabiei was identified. Treatment with ivermectin at resulted in complete cure of the dog. Meanwhile the humans were treated with sulfur ointment. Two weeks after this event the brother of the first dog, arrived in our laboratory where it was diagnosed with sarcoptic mange, too. Veterinary check of animal movement should target not only internal and infectious diseases but also the parasitic diseases, the latter ones generally being overlooked by the specialists. The release of health certificate covering animal health should be seen as an issue of utmost importance and responsibility for the veterinarians.

DOINA PĂTRAŞCU
DSVSA Caraş-Severin

Gasterophilus are parasites from Phylum Arthropoda, Order Diptera that are parasitic in the larval stage in the digestive tract of horses. Out of the 99 horses from Caraş-Severin county, killed mid-July 2006 in the slaughterhouse SC. Carolimp from Ucea of Jos, Brasov County, 89 had infections with Gasterophilus spp., corresponding to a prevalence of 89.89%. In horses from Caraş-Severin, subject to our study larvae of two species of Gasterophilus were identified: Gasterophilus intestinalis and Gasterophilus nasalis. The first species was found in the stomach, the secretory mucosa and always in the anterior part of the “margo plicatus” area. The second species was identified only in the duodenum, in the area immediately neighboring pyloric orifice. The extensitivity of infection with Gasterophilus intestinalis was 70.8% while the extensitivity of infection with Gasterophilus nasalis was 29.22%, G. intestinalis being always the predominant species. In the case of infections with only one species, G. intestinalis had an incidence of 7.85%, while G. nasalis had an incidence of 3.37%. In the remaining cases, the two species where found together. The total number of larvae counted was 12 226, intensitivity of infection was high, being on average 137.3 larvae/horse. “Nests” of larvae of the species Gasterophilus intestinalis were made up of between 17 and 107 individuals and those of Gasterophilus nasalis between 6 and 93, with an average of 36 larvae/nest for G. intestinalis and 34 larvae/nest for G. nasalis. The differences in extensitivity of infection between the sexes were not significant, being 68.42% for males and 64.28% for females. On age, horses of 1-5 years had the highest percentage of individuals with the highest intensivity (72.72%), followed closely by horses over 10 years (71.4%). Only in the 6-10 year category, the percentage of individuals who had over 50 each larvae is lower (58.69%). Lesions induced by Gasterophilus intestinalis larvae are in the crater-form formations with a diameter of about 0.3-0.5 mm and a depth of about 0.4 mm. They corresponded to the places where the larvae were fixed. Congestions, hemorrhagic lesions and even erosions were observed on the duodenal mucosa where the “nests” of parasites were big occupying large areas of mucosa.

D. PANAITESCU
Academia de Ştiinţe Medicale

The studies on Echinococcus granulosus protoscolices structure were made for many years using vital staining with methylene blue 1‰. This method allowed a rather good visualization of structural elements of this parasitic component which occurs during the biological cycle of the cestode Echinococcus granulosus. Examination by means of the microscopy with interferential light or polarised light permites visualization of living protoscolex making evident in different wavelengths from infra-red to ultraviolet a series of structures that were invisible by vital usual staining. It was also possible to watch among other things the structure of the cells with vibratile flag, the structure of the future proligerous membrane and many other elements unseen by the best vital staining with methylene blue. Our researches were refered as well to invaginated protoscolex as to evaginated protoscolex, where the second segment it is of the greatest importance in the development of the future hydatid cyst.

M. OVCHARENKO, P. WRÓBLEWSKI
Institutul de Parazitologie “W. Stefański”, Academia Poloneză de Ştiinţe, Warszawa, Polonia

Microsporidia are particularly common parasites of amphipod crustaceans, in which they are transmitted both horizontally and vertically. Cucumispora dikerogammari (Ovcharenko and Kurandina, 1987) cause emerging diseases of Ponto-Caspian invasive amphipod Dikerogammarus villosus. They has recently colonized most large rivers of Western and Central Europe. Infected individuals of Dikerogammarus villosus were sampled from the Bug River, Eastern Poland (N 52º27′30.79″; E 21º01′04.52″). The infected tissues were stored in a refrigerator at +4 ºC until the moment of experimental infection. Eighty uninfected individuals of invasive Dikerogammarus villosus (40) and native Gammarus pulex (40) were selected for the experimental infection. Pieces of the body of infected individuals were then provided as the only source of food for 3 days. After this exposure period, animals were fed with dry fish food pellets for 60 days. All living gammarids were then killed and dissected under a light microscope for examination of microsporidian infection. All of Dikerogammarus villosus survived until the end of the experiment demonstrated clearly visible symptoms of infection. None of Gammarus pulex displayed any evidence of infection.

B. NIŢU*, CORINA CIOBANU**, TICUŢA NEGREANU-PÎRJOL*, RALUCA NIŢU**, PARASCHIVA ENOPIAN**, E. SCARLAT***, ELENA GORUN**
*Facultatea de Farmacie, Universitatea ”Ovidius”; **Pro Diagnostic SRL; ***Nexus Alpha SRL, Constanţa

In this study, we sought to evaluate the antiparasitic effects of complex compounds of Zn, Cu and Ag with chlorhexidine. Amoebicidal effect of chlorhexidine is documented in the literature. Salts of metals Zn, Cu and Ag were noted by antimicrobial and antifungal effects. Because of lack of a reference of parasites as we for bacteria or yeasts we had to choose a protozoan, Trichomonas intestinalis, to test, then identify and isolate him in the stool of a patient with enterocolitis. Of this sample were prepared 25 slides with test protozoan as follows: 22 for complex compounds and one for the two standards, 96% ethanol and chlorhexidine dihydrochloride. On the last slide we tested the solid chlorhexidine effect as a mean of validation. On each slide have been identified vegetative forms of mobile protozoan (trophozoites). We added one drop of aqueous solution of complex compound to be tested and observed its effect in a period of 10 minutes. The selection criterion chosen was that after 1 minute the mobile protozoa no longer are observed. Although in all cases was observed protozoan movements cease (immobilization), it has not always produced fast enough. Solid chlorhexidine has also led to parasite immobilization, indicating its effectiveness in specified test conditions. There were selected 7 complex compounds, which have proved most effective in trophozoites immobilization. They will undergo further tests in order to obtain commercial products.

GABRIELA NICOLESCU*, VALERIA PURCĂREA-CIULACU*, AL. F. VLADIMIRESCU*, L. F. PRIOTEASA*, ELENA FALCUŢĂ*, ELENA CLAUDIA COIPAN*, GABRIELA DUMITRESCU**, DIANA POPESCU**, AURORA ALEXSE*
*INCDMI ”Cantacuzino”; **Centrul de Cercetări Medico-Militare, Bucureşti

The main diseases transmitted by mosquitoes in Romania are malaria and West Nile virus (WNV) infections. Re-emergence of WNV human infections and the increased risk of malaria re-emergence represented the result of the global environmental changes, especially climatic ones, in the last years. The evaluation of the risk level of mosquito-borne diseases in different ecosystems by multidisciplinary investigations using entomological, immunological, viral and molecular biology techniques was performed. The investigation of WNV circulation in its transmission cycles in natural and anthropic ecosystems showed the significant values of the seroprevalence of the antibodies against this virus in domestic and wild birds (main vertebrate hosts) and horses (tangential hosts). The virus was detected in Coquillettidia richiardii and Culex pipiens mosquitoes and was isolated from Culex pipiens females. The investigations showed the intensive and permanent enzootic circulation of WNV on extended territories over all the country and the permanent risk of its transmission to humans. The anophelism is the essential element of the risk of malaria re-emergence in the present circumstances of the increase of the number of imported malaria cases. The anophelism of the former malaria endemic areas is now represented by high abundant populations of the potential vectors of maculipennis group (Anopheles atroparvus, An. maculipennis, An. messeae and An. daciae). Their distribution is still registered in all the areas corresponding to the historical scheme of “malaria stratification” in Romania. The permanent surveillance of mosquitoes and implementation of integrated control programmes are adequate decisions in the risk areas of re-emergence of mosquito-borne diseases.

Work supported by funding from FP6 of EU (Contract No. 010284/2004) and CEEX and PN II Programmes of the Ministry of Research, Romania (Contracts No. 92 / 2006 and No. 42-125/2008).