Inhibitory activities of essential oils against Babesia canis.

The in vitro anti-Babesia canis activities of nine essential oils were investigated. Among the tested essential oils Achillea millefolium, Eugenia caryophyllus and Citrus grandis were the most active (IC50 values of 51.0, 60.3 and 61.3 µg/mL, respectively). The oils from Abies sibirica, Rosmarinus officinalis, Eucalyptus globulus, Cinnamonum zeylanicum, Mentha piperita and Pinus sylvestris were less active (IC50 values of 134.3, 237.3, 239.3, 367.9, 837.5 and 907.3 µg/mL, respectively). The results support the concept that some essential oil constituents may be useful in the clinical management of babesiosis.

The influence of bilberry fruit on memory and the expression of parvalbumin in the rat hippocampus.

Therefore, the aim of the present study was to evaluate the possible effect of bilberry fruit (Vaccinium myrtillus L.) supplement in a daily diet on the cognitive behaviour of the rats and the expression of paravalbumin (PV) in populations of hippocampal neurons. It has been postulated that the antioxidants present in bilberry fruit may act as neuroprotective factors playing also a significant role as memory enhancements. Forty Wistar rats with a similar average body weight (460 ± 0.4 g) were divided into four groups (n=10 per group). The control group received standard feed (210 g/week), whereas animals of experimental groups received standard feed supplemented with bilberry (per os) at consumed doses of 2 g (group I), 5 g (group II), and 10 g/kg b.w./ /day (group III). After three months of feeding with bilberry, the modified elevated plus-maze test (mEPM) was performed. After 32 weeks of feeding, brains were collected and PV-immunoreactive (ir) neurons were immunohistochemically visualized. In the modified elevated plus-maze test, transfer latency examined 2 h and 24 h after the acquisition session was significantly shorter (p⟨0.05) in the group II in comparison with the control group. In CA1 and CA2/CA3 hippocampal fields as well as dentate gyrus of all experimental groups, a significant (p⟨0.05) decrease in number of PV-ir neurons were found. In relation to the control group, the mean subpopulation of PV-ir neurons found in groups II and III were significantly reduced. The subpopulations of PV-ir neurons found in DG of all experimental groups were significantly reduced in comparison to the control. In conclusion the in the present paper we demonstrated a relationship between the diet rich in a bilberry fruit and process of memory as well as numbers of calcium- binding protein-expressing hippocampal neurons. Our results may be source of basic knowledge for further research aiming at neuroprotective role of the bilberry fruit.

The concentration of urea in hemolymph as a marker of health in Lissachatina fulica and Cornu aspersum edible snails - a preliminary study.

Edible snails are kept in farms in many countries worldwide. As farm animals, they are an object of interest of veterinary studies and applied biology. There is a large demand for tests which would help identify their health and well-being. The objective of this study was to assess the usefulness of determining the concentration of urea in hemolymph as a marker of health of the Lissachatina fulica and Cornu aspersum edible snails. The observation covered snails from four farms marked from A to D, in which numerous deaths (farm A) and decreased body weight gain (farms B and C) were observed. In experimental farm D we observed a group of snails subjected to stress and a control group maintained in correct conditions. High concentrations of urea were found in the hemolymph of all farm animals from farms A, B and C, as well as in those subjected to food deprivation in farm D (on average from 96 mg/dl in farm D to 320 mg/dl in farm A). On the other hand, in controls from group D, the concentration of the parameter in question was much lower (⟨ 2.0 mg/dl). The results obtained indicate that the urea concentration is a non-specific marker of pathological conditions in snails, and that the continuous monitoring of this parameter makes it possible to demonstrate irregularities in farming and introduce appropriate and early measures to eliminate such disturbances.

In vitro antioxidant and antibabesial activities of the extracts of Achillea millefolium.

Despite many phytochemical and pharmacological investigations, to date, there are no reports concerning the antibabesial activity of extracts of A. millefolium against B. canis. This study was aimed at investigating the biological activities of A. millefolium against the Babesia canis parasite and to identify its chemical ingredients. The water (WE), ethanol (EE) and hexane/acetone (H/AE) extracts of plant aerial parts were screened for total phenolic content (TPC), total flavonoid compound (TFC), DPPH free radical-scavenging activity and its antibabesial activity assay. In this study, imidocarb diproprionate was used as a positive control. The H/AE and EE extracts were analysed using gas chromatography-mass spectroscopy (GC-MS). In the EE extract, the main compounds were 17.64% methyl octadec-9-ynoate, 16.68% stigmast-5-en-3-ol(3α,24S) and 15.17% hexadecanoic acid. In the H/AE extract, the main compounds were 34.55% 11-decyldocosane, 14.31% N-tetratetracontane, 8.22% ß-caryophyllene, and 7.69% N-nonacosane. Extract of EE contained the highest content of phenolics followed by H/AE and WE. The concentration of flavonoids in EE, H/AE and WE extracts showed that TFC was higher in the EE samples followed by H/AE and WE. The antioxidant activities were highest for AA, followed by EE, WE and H/AE. The antibabesial assay showed that the WE, EE and H/AE extracts of A. millefolium were antagonistic to B. canis. At a 2 mg/mL concentration, it showed 58.7% (± 4.7%), 62.3% (± 5.5%) and 49.3% (± 5.1%) inhibitory rate in an antibabesial assay, respectively. Considering these results, the present findings suggest that A. millefolium extracts may be a potential therapeutic agent and that additional studies including in vivo experiments are essential.

Study on establishing normal ranges of chosen biochemical parameters of haemolymph of Cornu aspersum maxima and Cepaea nemoralis gastropods.

The aim of the study was to establish normal ranges for chosen biochemical parameters of haemolymph of snails (Gasropoda: Mollusca), in the light of the use of these animals as experi- mental models in various types of studies. The study was conducted on 100 specimens of Cornu aspersum maxima (CAM) and 100 specimens of Cepaea nemoralis (CN). The haemolymph col- lected from the animals was analysed using colorimetry to assay aspartate transaminase (AST) activity, alanine transaminase (ALT) activity, amylase activity and the concentrations of urea and triglycerides. In the further part of the study, the influence of administering doxycycline with feed on the change of AST and ALT activity in snail haemolymph has been studied. The normal values established for CAM are as follow: AST activity: 26-38 u/l, ALT activity: 0-11 u/l, amylase activity 9-16 u/l, concentration of urea: 3-6 mg/dl, concentration of triglycerides: 16-20 mg/dl. For CN, the following data have been obtained: AST activity: 30-80 u/l, ALT activity: 0-15 u/l, amylase activity 12-15 u/l, concentration of urea: 5-8 mg/dl, concentration of triglycerides: 18-24 mg/dl. It has been shown that doxycycline presents a high workload on the hepatopancreas of snails, which is reflected by a statistically significant (p ⟨ 0.05) increase of AST and ALT activity in the haemolymph of the specimens which obtained doxycycline in feed, as compared to the groups with antibiotic-free feed. The haemolymph activity of both studied parameters increased together with study time and tetracycline administration time.

The Impact of Caregivers' Anxiety on Patients' Anxiety before Fast-Track Knee Arthroplasty.

PURPOSE OF THE STUDY Anxiety may have negative repercussions on the surgery including poorer outcomes. On the other hand, the majority of patients reporting preoperative anxiety fear not receiving enough attention from a caregiver. In patients undergoing fast-track knee arthroplasty, we determined the relationship between patients' preoperative anxiety and the anxiety the patient's caregiver. We also analyzed the influence of selected psychosocial and demographic variables on the relationship between caregivers' and patients' anxiety. MATERIAL AND METHODS We conducted a prospective, descriptive study in which baseline assessments of patients scheduled to undergo fast-track total knee arthroplasty between 1st November 2014 and 30th April 2015 were compared with those of their caregivers. Patients were recruited from a large teaching hospital through the orthopedics joint replacement clinic. Information on sex, age, body mass index (BMI), educational status, employment status, marital status, and living status was recorded for all patients. Patients and their caregivers completed the Spielberger State-Trait Anxiety Inventory. Baseline trait anxiety was assessed with STAI scores in the initial interview, 2 weeks before hospitalization, and state anxiety was assessed the day before the surgery. The patients' caregivers were contacted during a scheduled postoperative clinic visit and asked to complete the STAI and to provide information on their age, degree of consanguinity with patient, and living status. RESULTS The mean age was 66.4 years for the 118 patients and 55.7 years for the 93 caregivers. In male caregivers, caregiver anxiety and patient anxiety were positively related but not statistically so, and in women was not significant. In male patients, a relationship between caregiver's anxiety and patient's anxiety was positive, although not statistically significant, and in women was neither present nor significant. DISCUSSION Given the widespread impacts of anxiety before knee arthroplasty, it is critical for surgeons to gain a better understanding of how to identify and reduce preoperative anxiety in operated patients. We found that male sex among caregivers was associated with more preoperative anxiety among patients than was female sex and that male patients more quickly accepted anxiety from their caregivers than did female patients. CONCLUSIONS Anxious male caregivers appear to impart their anxiety to male patients but not to female patients. The anxiety of unrelated caregivers is associated with low preoperative anxiety among patients. Preoperative interventions should focus on caregivers, especially male caregivers, and to related caregivers to help patients cope with anxiety before knee arthroplasty. Key words: knee arthroplasty. knee replacement. fast track, anxiety, caregiver, preoperative stress.

Application of Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry for Identification of Coagulase-negative Staphylococci Isolated from Milk of Cows with Subclinical Mastitis.

The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81.8%). The most frequent cause of subclinical mastitis was found to be Staphylococcus sciuri (39%), while S. vitulinus was detected in 15% of the milk samples. The results obtained indicate that MALDI-TOF MS can be used for the identification of CNS isolated from bovine mastitis as a method supplementary to biochemical tests.

The concentrations of inflammatory cytokines and acute-phase proteins in the peripheral blood and uterine washings in cows with pyometra.

The development of pyometra in cows depends largely on the state of local immunity of the uterus. The objective of the study was to evaluate the concentration of the following proinflammatory cytokines: tumour necrosis factor (TNF-α) and interleukin-6 (IL-6); anti-inflammatory cytokine: interleukin-10 (IL-10); and acute-phase proteins (APPs): haptoglobin (Hp) and serum amyloid A (SAA), in serum and uterine washings in cows with pyometra and healthy animals. The study was performed on 20 cows divided into two groups based on the results of cytological and ultrasonographic tests: a pyometra and a healthy group (10 cows per group). Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines, TNF-α, IL-6, IL-10 and APPs - Hp and SAA, in the study material were determined by ELISA. The results showed that the values of TNF-α, IL-6, IL-10 as well as SAA and Hp were significantly higher in serum of cows with pyometra compared to controls (p < 0.001). The uterine washings had significantly higher levels of IL-6, IL-10, and Hp in pyometra cows compared to the control (p < 0.001). Our results indicate that it is possible to monitor the course of pyometra in cows based on the evaluation of the concentration of cytokines and Hp in the serum and uterine washings. Simultaneous evaluation of selected indicators of antagonistic interaction can be helpful in determining the current status of local immunity of the uterus. On this basis, it could be possible to properly select an adjunctive therapy in the form of immunomodulating preparations.

Effect of adding one 15-minute-walk on the day of surgery to fast-track rehabilitation after total knee arthroplasty: a randomized, single-blind study.

BACKGROUND: Earlier and more intensive physiotherapy exercise after total knee arthroplasty (TKA) enhance recovery, but the best combination of intensity and duration has not been determined. AIM: To determine whether adding a single, 15-minute walk on the day of surgery to a fast-track rehabilitation protocol would reduce knee pain and improve knee function after TKA. DESIGN: A randomized single-blind study. SETTING: Inpatient. POPULATION: Patients with primary osteoarthrosis after TKA. METHODS: Patients undergoing TKA were randomly assigned to a standard, fast-track rehabilitation protocol consisting of a single, 15-minute walk with a high-rolling walker 4 to 6 hours after recovery from spinal anesthesia or to an intensive protocol, in which patients took a second 15-minute walk at least 3 hours after the first, only on the day of surgery. Outcomes were pain measured on a visual analog scale, Knee Society's (KSS) clinical and functional scores, Oxford knee scores, and Spielberger State-Trait Anxiety Inventory scores. Patients were blinded to group assignment. Since most data were non-normally distributed non-parametric tests were used. Groups were compared with Mann-Whitney U test (for continuous variables). Association between continuous variables was evaluated with Spearman`s rank correlation coefficient. Chi-square or Fisher's exact test was used to assess differences in categorical variables. RESULTS: Of 86 patients assessed for eligibility, 66 were randomly assigned. The 31 evaluable patients on the intensive protocol (mean age, 68 years; 18 women) did not differ significantly from the 31 (mean age, 70 years; 20 women) on the standard protocol on any baseline characteristic or on any outcome measure on any day. On the second postoperative day, pain while walking dropped from a mean of 6.1 to a mean of 4.9 in the intensive group and from 6.4 to 5.4 in the standard group. Results for pain at rest were 3.3 to 2.2, respectively, for the intensive group and 4.0 to 3.0 for the standard group. At 2 weeks, pain at rest was 2.8 in both groups, and pain while walking was 3.0, respectively, for the intensive group and 3.4 for the standard group. At 2 weeks, mean (SD) KSS clinical and KSS function scores were, respectively, 74.9 (12.5) and 51.6 (16.2) in the intensive group and 71.2 (14.3) and 46.3 (16.1) in the standard group. Older age correlated with decreasing knee function (rS=-0.43, P<0.001), and less knee flexion correlated with preoperatively higher state anxiety (rS=-0.37, P=0.005) and trait anxiety (rS=-0.29, P=0.027). The study is limited by its small sample. The fast-track program was not in line with the best available evidence following knee arthroplasty, because patients did not undergo such treatment as NMES. Finally, the intervention itself was modest. CONCLUSION: Adding an additional 15-minute walk to a fast-track rehabilitation protocol did not increase pain, but neither did it improve functional recovery. CLINICAL REHABILITATION IMPACT: A 15-minute walk immediately after recovery from spinal anesthesia did not increase pain in patients with TKA. More intense exercise during this period might improve functional recovery without increasing pain.

Diagnosis of the Encephalitozoon cuniculi infections in pet rabbits with neurological symptoms.

The purpose of the study was the in vivo diagnosing of E. cuniculi invasions in pet rabbits with neurological symptoms using the Real-Time PCR, and determination of the rate of invasion, in this group of animals. The study involved 103 pet rabbits with neurological symptoms. Parasitic invasions were diagnosed using Real-Time PCR. The DNA of the parasites for molecular tests was isolated from the urine of the diseased animals. Out of the 103 tested DNA samples, the presence of the E. cuniculi genetic material was detected in 27 samples (26.21%). The melting temperature (Tm) of all products was 77.5 degrees C. The presence of parasitic DNA in the urine of 26.21% of examined animals indicates that E. cuniculi infections occur widely in pet rabbits in Poland and are a significant cause of neurological disorders in those animals.

The identification of Anaplasma spp. isolated from fallow deer (Dama dama) on a free-range farm in eastern Poland.

The aim of the present study was to investigate the occurrence of Anaplasma spp. in group of 50 fallow deer (Dama dama) from free-range farm in eastern Poland and determine what species of Anaplasma could infect these animals based on PCR gene sequencing. The PCR technique revealed the presence of 16S RNA Anaplasma spp. genetic material in the blood of 7 out of 50 examined animals. The sequences of the PCR products obtained showed a 100% homology with each other and 100% homology with GU 183908 sequence of A. phagocytophilum, isolated in our earlier study from a horse with clinical form of anaplasmosis. Here, we report the first molecular evidence of Anaplasma spp. among naturally infected fallow deer in eastern Poland.

Phylogenetic analysis of canine parvovirus CPV-2 strains and its variants isolated in Poland.

Canine parvovirus disease appeared in the world and in Europe during the second half of the 1970s. Over the course of 40 years the original CPV-2 strains mutated and variants 2a, 2b and 2c appeared. Their appearance is connected with specific amino acid changes, mainly in the capsid protein VP2. Strains isolated by the authors were adapted for in vitro cell culture. Phylogenetic analysis revealed differences between strains isolated in Poland in 1982-1985 and in 1995-2009. Strains from the 1980s were shown to belong to variant CPV-2a (11 strains) and variant 2b (2 strains), while no fundamental differences were found among the genetic profiles of the strains from 1995-2009, which were classified as belonging to variant 2c.

[Pregnancy and delivery in women with thrombocytopenia]. / Ciaza i poród u kobiet z maloplytkowoscia.

Maternal immune thrombocytopenia is a frequent finding in pregnancy and is most commonly asymptomatic and clinically benign. 8 pregnant women with thrombocytopenia between January 1995 and December 2000, in Clinic of Obstetrics and Gynecology in Katowice-Ligota, were evaluated. 7 patients had idiopathic thrombocytopenia purpura and one (31 years old) had cirrhosis, splenomegaly, thrombocytopenia and elevated liver enzymes. 6 cases was steroid-treated with initial platelet count 4,000 per cubic millimeter to 115,000 per cubic millimeter after treatment. Alloimmune thrombocytopenia is a serious fetal disorder, resulting from platelet-antigen incompatibility between the mother and fetus. Among all newborns only one had thrombocytopenia, platelet count 45,000 per cubic millimeter, without neonatal intracranial hemorrhage. 6 women had cesarean section and 2 patients delivered naturally.