Biochemical and histomorphological findings in Swiss Wistar rats treated with potential boron-containing therapeutic - K2[B3O3F4OH].

BACKGROUND: Boron and boron containing compounds are known for their biological and protective roles being non-toxic and non-mutagenic in low concentrations. Male rats were exposed to halogenated boroxine (HB), dipotassium-trioxohydroxytetrafluorotriborate K2[B3O3F4OH], a potential new boron-containing therapeutic, aiming to determine concentrations with no adverse effects on selected serum biochemical parameters and histomorphological features. METHODS: HB was prepared by reacting potassium hydrofluoride (KHF2) with boric acid in molar ratios 2:3 at room temperature and its primary structure contains 4 fluorine atoms substituted in 6-membered ring. In concentrations of 10, 25, 35 and 45 mg/kg, HB was administered intraperitoneally as a single dose. Biochemical parameters were observed 24 and 96 h following the treatment. Effects of HB on biochemical blood parameters were also observed 24 h following continuous nine days application in concentrations of 10 mg/kg intraperitoneally and 50 mg/kg per os. Histomorphological observation of kidneys, liver, spleen, lungs and heart was performed for all treated animals. RESULTS: Administration of single high dose of HB (35 mg/kg-45 mg/kg) effected high levels of urea and creatinine, which indicated renal injury that appeared to be temporary. Possible cause of concern is pancreatic injury indicated by elevated levels of serum amylase in the groups of animals that received the highest dosages of the substance. Histopathological examination of selected tissues revealed mild to moderate lesions in the kidneys and livers associated with administration of HB. CONCLUSION: Observation of biochemical serum parameters or histopathology of examined tissues revealed no adverse effects of HB either after the administration of single dose lower than 35 mg/kg or following repeated administration at 10 mg/kg. These dosages should be further considered for potential therapeutic applications.

Fatal visceral leishmaniosis in a dog caused by Leishmania infantum in Bosnia and Herzegovina: A case report.

Canine leishmaniosis (CanL) caused by Leishmania infantum, is a zoonotic vector-borne disease endemic in the Mediterranean region. Here, we report a molecularly confirmed case of fatal CanL caused by L. infantum in the south of Bosnia and Herzegovina where epidemiology data are scarce. A 2.5-year-old, male golden retriever presented with a history of lethargy, prostration, and anorexia. Clinical examination revealed pale mucosae membranes, reduced capillary refill time, anuria, and ulcerated oral mucosae and skin of the legs. Complete blood count discovered severe non-regenerative, normocytic and normochromic anemia. Biochemistry profile showed hyperglycemia, hypoalbuminemia, hypercholesterolemia, increased potassium, and considerably elevated creatinine, urea, and phosphorus. Rapid Leishmania SNAP test was negative, as well as the serum neutralization test for leptospirosis. At necropsy, mildly enlarged and firm yellow to tan kidneys were the most prominent lesions. Macrophages laden with amastigotes in bone marrow, liver, spleen, kidneys, lymph nodes and the skin were seen in histopathology. Molecular testing by PCR and sequencing (cpb gene) confirmed and identified the pathogen as L. infantum. This study highlights the lack of key measures necessary to undertake the proper control of this important zoonosis in the country. Nationwide epidemiologic study on CanL and its vector(s), along with adoption and establishment of proper diagnostic approach with quantitative serologic and molecular methods in place are warranted.

Fatal pulmonary cysticercosis caused by Cysticercus longicollis in a captive ring-tailed lemur (Lemur catta).

Here we describe fatal pulmonary cysticercosis caused by Cysticercus longicollis, the larval stage of Taenia crassiceps in a 15-year-old female ring-tailed lemur (Lemur catta) from Sarajevo Zoo. After sudden death, the lemur was subjected to necropsy and large multicystic structure, subdivided with fibrous septa and filled with numerous translucent, oval to ellipsoid bladder-like cysts (cysticerci), almost completely replacing right lung lobe was observed. In addition, numerous free and encysted cysticerci were found in the thoracic cavity. Histopathology revealed connective tissue outlined cavities that compress lung parenchyma. Each cavity contained several thin walled cysticerci with single inverted protoscolex, one or more suckers and rostelum with two rows of hooks. In many of the cysticerci one or several exogenous buds of daughter cysticerci were observed. Based on morphology and microscopic appearance the parasite was identified as C. longicollis. Subsequent molecular analysis and sequencing confirmed presumptive diagnosis. To our knowledge, this case represents the first report of T. crassiceps and cysticercosis caused by C. longicollis in Bosnia and Herzegovina.

Hepatozoon silvestris sp. nov.: morphological and molecular characterization of a new species of Hepatozoon (Adeleorina: Hepatozoidae) from the European wild cat (Felis silvestris silvestris).

Based on morphological and genetic characteristics, we describe a new species of Hepatozoon in the European wild cat (Felis silvestris silvestris), herein named Hepatozoon silvestris sp. nov. The study also provides the first data on the occurrence of H. felis in this wild felid. Hepatozoon meronts were observed in multiple cross-sections of different organs of four (44%) cats. Additionally, extracellular forms, resembling mature gamonts of Hepatozoon, were found in the spleen and myocardium of two cats. Furthermore, tissues of six animals (67%) were positive by PCR. Hepatozoon felis was identified infecting one cat (11%), whereas the 18S rRNA sequences of the remaining five cats (56%) were identical, but distinct from the sequences of H. felis. Phylogenetic analyses revealed that those sequences form a highly supported clade distant from other Hepatozoon spp. Future studies should include domestic cats from the areas where the wild cats positive for H. silvestris sp. nov. were found, in order to investigate their potential role to serve as intermediate hosts of this newly described species. Identification of its definitive host(s) and experimental transmission studies are required for elucidating the full life cycle of this parasite and the possible alternative routes of its transmission.

Pearsonema plica (Capillaria plica) infection and associated urinary bladder pathology in red foxes (Vulpes vulpes) from Bosnia and Herzegovina.

Pearsonema plica is a widely distributed nematode parasite that occurs in the urinary tract of various domestic and wild carnivores. The aim of this study was to investigate the occurrence and geographical distribution of P. plica and associated urinary bladder pathology in 112 red foxes (70 males, 42 females; 87 adults >1 year, 25 juveniles <1 year) from six different geographical regions in Bosnia and Herzegovina. The urinary bladders of the red foxes were subjected to gross examination and histopathology. Urine content (n = 40) and mucosal smears (n = 71) of the urinary bladders were examined microscopically for the presence of P. plica. Overall, adults and eggs of P. plica were detected in 65 (58.0 %; 95% CI 48.9-67.2%) of the foxes. Out of the positive foxes, 42 were males (64.6%) and 23 females (35.3%). According to age, 49 adults (75.3%) and 16 juveniles (24.6%) were positive. There were no statistically significant differences in the infection prevalence between the geographical regions (p = 0.701), sex (p = 0.693), or age (p = 0.646) of the host. Also, no significant differences in the prevalence of parasites in urine content (48.7%; 20/41) and mucosal smears (63.3%; 45/71) were observed (p = 0.165). Eosinophilic cystitis characterized with mild to severe infiltrates of eosinophils in the propria of the bladder mucosa accompanied by hyperemia and edema was observed in 36 examined foxes, 24 of which were P. plica positive. Parasites attached and embedded into the mucosa and free in the lumen were recorded in both cystitis positive and negative foxes. Beside clear numerical differences, the influence of P. plica infection on the occurrence of cystitis was not significant (p = 0.309). The results of this study give the first insight into the distribution of P. plica infection among the red fox population in Bosnia and Herzegovina. Observed microscopic changes may contribute toward a better understanding of pathology caused by this widely distributed parasite in free-ranging red foxes.

Highly pathogenic avian influenza virus subtype H5N1 in mute swans (Cygnus olor) in Central Bosnia.

In order to determine the actual prevalence of avian influenza viruses (AIVs) in wild birds in Bosnia and Herzegovina, extensive surveillance was carried out between October 2005 and April 2006. A total of 394 samples representing 41 bird species were examined for the presence of influenza A virus using virus isolation in embryonated chicken eggs, PCR, and nucleotide sequencing. AIV subtype H5N1 was detected in two mute swans (Cygnus olor). The isolates were determined to be highly pathogenic avian influenza (HPAI) virus and the hemagglutinin sequence was closely similar to A/Cygnus olor/Astrakhan/ Ast05-2-10/2005 (H5N1). This is the first report of HPAI subtype H5N1 in Bosnia and Herzegovina.

Topical treatment of standardised burns with herbal remedies in model rats.

Experimental studies of burns require the use of different animal models. The aim of this work was to establish experimental model of thermal injuries and to evaluate the effects of topical agents on healing of the burn wounds. Forty female Wistar rats were randomly classified in 4 groups and isolated for 2 weeks before the onset of experiment. Animals were primarily anaesthetized with pentobarbital-sodium and then shaved (skin area of their back with diameters 5 cm x 5 cm). A round metal stamp with contact area of 5 cm2 and total weight of 100 g was heated up to 80 degrees C and then applied without additional pressure on the depilated skin of the back for 14 seconds. This procedure produced a standardized burn wound. Induced burn wounds were immediately drowned in the 4 degrees C- water for 3 s in order to maintain microcirculation. After the inducement of thermal injures, all rats were treated with 1% silver sulfadiazine cream, herbal topical preparations or were not treated at all. Burn wounds were treated twice a day until the healing completion. The result of treatment application was a significant reduction of burn wound diameters. Herbal topical preparations expressed positive therapeutic effects on the parameters of burn wounds. The efficiency of silver sulfadiazine cream in burn wound healing was significantly more expressed in comparison to healing process in control group of animals (p < or = 0,001). We conclude that herbal topical preparations efficiently caused the completion of burn wound healing process without scar formation.

Nitric oxide in gentamicin-induced acute tubular necrosis in rats.

Gentamicin is still widely used in clinical practice in spite of its renal toxicity. The role of nitric oxide (NO) in that process is not completely elucidated. The aim of this study was to investigate the relationship between plasma level of NO and the histopathological changes of kidney in acute tubular necrosis (ATN) induced by gentamicin in rats. Study was carried out in Albino-Wistar rats, both sexes (n=16), average body weight 200-250 g. divided in two equal groups: control and gentamicin group. The control group was injected with 0.9% NaCl i.p. and gentamicin group was injected with gentamicin in the dose of 80 mg/kg/day i.p. in a period of 5 consecutive days. NO plasma level was determined by the production of nitrates and nitrites using classical colorimetrical Griess reaction. Kidney specimens were stained with hematoxylin-eosin (H-E) and Periodic acid-Schiff (PAS) stain. Semiquantitative histological analysis was used for the evaluation of the level of kidney damage. Both, the plasma NO level and the level of kidney damage were statistically higher in rats with gentamicin-induced ATN in comparison to the control group. In spite of that the correlation between plasma NO level and the level of kidney damage was not found. The rise of plasma level NO in gentamicin induced ATN in rats could possibly indicate on the role of NO in renal damage caused by gentamicin.