Evaluation of vasoactivity after haemotoxic snake venom administration.

The chick chorioallantoic membrane (CAM) is a widely used model in medical research and fulfils the requirements laid out in the 3 R's. The CAM, contains a dense network of blood vessels, essential for embryo development but also makes the chick embryo an invaluable resource for the study of angiogenesis and the haemotoxicity of substances. Non-neurotoxic snake venom is responsible worldwide for numerous deaths but also has been found to have a vast range of medicinal benefit. This study combines evaluating the time of onset and intensity of effects of three whole viper venoms (Bitis aritans, Crotalus viridis, Agkistrodon contortrix) at varying concentrations. They were applied onto the CAM, using the Luepke grading system as one method of determining their rapid irritation potential. Regarding the principles of 3 R's, this method helps to evaluate the haemotoxic effect of venom as an alternative method to the rodent tests. The information provided from these results can be used as a rapid tool for both medical management of snakebite wounds and the potential use of snake venom in medical treatments. Then, Luepke grading system can help to evaluate the haemotoxic effect of venom in combination with other appropriate methods.

DNA detection of Toxoplasma gondii in sheep milk and blood samples in relation to phase of infection.

Toxoplasma gondii is a worldwide parasite that is important both for veterinary medicine (economic losses in the herd) and for public health (immunocompromised patients and pregnant women). An important source of Toxoplasma infection in humans is consumption of contaminated meat and milk (undercooked meat and unpasteurized milk). Small ruminants are important in both milk and meat production throughout the world because of free-range husbandry. The purpose of our study was to detect the presence of T. gondii DNA in ewes' milk 1 month after the term, and to determine the relationship between the occurrence of this DNA in blood and milk based on the phase of infection. Using enzyme-linked immunosorbent assay (ELISA) the animals were divided into two groups (immunoglobulin M positive (IgM+), IgM-). With real-time polymerase chain reaction (PCR), T. gondii DNA was detected in seven milk samples (28%) and five blood samples (20%) of the IgM+ group (25 samples). In the IgM- group T. gondii DNA was detected in two milk samples (3.6%) out of 55 samples.

The occurrence of microsporidial infections and toxoplasmosis in Slovak women.

AIM OF THE STUDY: To determine the prevalence of antibodies to intracellular pathogens (Toxoplasma gondii, Encephalitozoon cuniculi, and Encephalitozoon intestinalis) in sera of Slovak women. MATERIAL AND METHODS: Enzyme-linked immunosorbent assay (ELISA) was used for the detection of specific IgG antibodies to Toxoplasma gondii, Encephalitozoon cuniculi, and Encephalitozoon intestinalis in 118 sera samples from Slovak women with different diagnoses. RESULTS: Women were divided into three groups based on the age. In group 1 (n = 26, age > 51), nine women tested positive for T. gondii, two for E. cuniculi, and one for E. intestinalis. The respective numbers in group 2 (n = 53, age range 36-50) were 18, three, and seven, and in group 3 (n = 39, age range 20-35) 13, two, and four. In an attempt to find the relationship between the clinical diagnosis and the prevalence of the three pathogens, we divided the study subjects into four groups based on the condition (diseases of the nervous, respiratory, and immune systems and pregnancy). A statistically significant relationship between the diagnosis and the prevalence of a pathogen was found (p < 0.001). In women with diseases of the respiratory system, antibodies against Toxoplasma gondii were most often detected, while infection with E. intestinalis was most commonly associated with nervous system diseases and the highest seroprevalence of E. cuniculi was found in women diagnosed with immune system disorders. CONCLUSIONS: In our study, the seroprevalence rates of the three infections in pregnant women were similar, showing no significant difference. Women with diseases of the respiratory system were most often seropositive for T. gondii. Our result is consistent with the known fact that in humans, toxoplasmosis may affect various organs of the body, causing diverse clinical signs. However, the respiratory system is commonly involved and pneumonia may result. The most common finding is a mild, flu-like illness that lasts a few days. The women with different types of immunodeficiency disorders had the highest seropositivity rates for E. cuniculi and the second most common infection in this group was toxoplasmosis. Both of these parasites are opportunistic and can cause serious problems in immunocompromised individuals (HIV-positives, organ transplant recipients, chemotherapy patients). The women diagnosed with the nervous system disorders were most commonly infected with E. intestinalis. The relationship between the disease and this pathogen is not clear, but E. intestinalis is responsible for various pathologies; it causes mainly disseminated infections and damage to the digestive tract. E. intestinalis is a parasite very close to E. cuniculi affecting either the nervous system or the respiratory tract depending on the type of host.

Bendiocarbamate toxicity in the chick embryo.

Carbamate pesticides generally possess low toxicity for warm-blooded vertebrates, but developmental data are scarce. We have therefore evaluated embryotoxicity of choline esterase inhibitor bendiocarbamate in the chick embryo. The pesticide was dissolved in 5% acetone in distilled water and a volume of 200 microl was administered over the embryo through membrana papyracea on embryonic days 2, 3, 4, 5, and 10. Sampling was performed on embryonic day 10, while the embryos treated on embryonic day 10 were sampled on embryonic day 17. The toxicity of bendiocarbamate was fairly low, and LD50 decreased with advancing development from 1 mg/ embryo on embryonic day 2 to 29 mg on embryonic day 5. Malformations in surviving embryos were observed rarely (< 3 %) and occurred in both control and experimental groups. There was a mild but statistically significant dose-dependent reduction in body weight, most pronounced in the treatment on embryonic days 5 and 10, but the maximum difference from controls was below 15 %. A small but not significant increase in the number of positive cells was observed in the eye, limb buds, and the central nervous system of embryos treated on embryonic days 3 and 4 and examined after supravital whole-mount staining with Lysotracker Red for apoptosis. In agreement with previously published studies in other vertebrate animals, we conclude that bendiocarbamate does not possess significant toxicity in the avian embryo.

Encephalitozoonosis in household pet Nederland Dwarf rabbits (Oryctolagus cuniculus).

The paper presents the results of examination of 32 domestically bred rabbits, the breed Nederland Dwarf of Oryctolagus cuniculus, for the presence of Encephalitozoon cuniculi microsporidian species. The results of serological tests for E. cuniculi in 32 rabbits are reviewed along with other follow-up studies of clinical cases. Blood samples were taken from 7 asymptomatic rabbits and 25 rabbits showing neurological and ocular signs suggestive of encephalitozoonosis. In the asymptomatic group, 5 out of 7 rabbits were seropositive (71%). 16 rabbits with clinical diseases showed neurological sings, including torticollis, circus-like movements, loss of weight; 6 of them also showed ataxia, anorexia, asthenia of hind-limbs and 3 showed ocular signs. All 25 rabbits were seropositive. The spores of E. cuniculi were isolated from the faecal samples or kidneys and brain of an animal and subsequently were used for DNA isolation and PCR analysis.