Prevalence of keds on city dogs in central Poland.

The aim of this study was to identify the species of ked infesting dogs in the cities of central Poland. A total of 510 dogs were observed between June and September 2015. The presence of keds was noted in 182 (35.7%) animals. Keds were more prevalent in female (38.0%) than in male (33.2%) dogs, and were more frequently found in animals younger than 1 year (46.2%) and in long-haired dogs (36.6%). The body areas most heavily colonized by keds were the groin (35.4%) and neck (21.4%). A total of 904 keds were isolated from dogs, including Hippobosca equina (Diptera: Hippoboscidae) (17.2%), Lipoptena cervi (Diptera: Hippoboscidae) (32.0%), and two species not previously encountered in Poland: Hippobosca longipennis (45.0%) and Lipoptena fortisetosa (5.9%). Hippoboscidae may act as vectors of pathogens and any shifts in their geographic range may lead to the spread of new diseases affecting animals.

Histopathological studies of the heart in three lines of broiler chickens.

The aim of this study was to analyse the morphological lesion pattern of the heart of broiler chickens (Cobb 500, Hubbard F15 and Ross 308) during fattening with no clinical signs of disease and to determine the most susceptible period for the occurrence of morphological lesions. The most frequently diagnosed lesions in each genetic line were degeneration of the fibres with vacuolation, congestion of cardiac muscle, oedema and vacuolisation of the Purkinje cells. The highest numbers of morphological lesions were observed on d 38, 31 and 10 of life. The lesions were most numerous in the septum, followed by the left and right ventricles. Ischaemic cardiomyocytes were also most numerous on d 38 of life and in the left ventricle. Overload of cardiac muscle, prolonged hypoxia and increasing body weight on d 38 are the likely reasons for the largest number of lesions and ischaemic fibres, which may lead to heart failure.

Detection of Pentatrichomonas hominis in dogs using real-time PCR.

Trichomonadidae family is a protozoan occurring in different animal species. It inhabits the gastrointestinal and urinary tracts. P. hominis is rarely found in faecal samples of dogs, and its identification and differentiation from other trichomonads by light microscopy are difficult. Methods of molecular biology are the most effective in this case, because they confirm the presence of the specific species in animal organisms, irrespective of the protozoan form. The aim of this study was to find P. hominis in selected dog kennels in North-Eastern Poland. Forty-one faecal samples of dogs from 7 dog kennels were examined. The occurrence of P. hominis in 5 faecal samples of dogs with no symptoms of diarrhoea was the first one to be confirmed in Poland.

A comparison of the efficacy and pharmacokinetics of ivermectin after spring and autumn treatments against Cyathostominae in horses.

The aim of the present study was to determine the efficacy of ivermectin against Cyathostominae infections and to describe the drug's pharmacokinetic parameters during two seasonal deworming treatments in horses. The study was performed on warm-blooded mares aged 3-12 years weighing 450-550 kg. A single bolus of an oral paste formulation of ivermectin was administered at a dose of 0.2 mg/kg BW in spring and autumn. Fecal samples were tested before treatment and 1, 2, 3, 4, 6, 10, 20, 30, 40, 50, 60, 75 days after treatment. Ivermectin concentrations in blood samples collected before treatment, 0.5, 1, 2, 3, 4, 6, 12, 24, 36 and 48 hours after treatment, and 3, 4, 6, 8, 10, 15, 20, 25, 30, 40, 50, 60 and 75 days after drug administration were determined by high pressure liquid chromatography. Drug absorption was significantly (p<0.05) slower (tmax: 21.89±11.43 h) in autumn than in spring (tmax: 9.78±8.97 h). Maximum concentrations (Cmax) of ivermectin in the blood plasma of individual horses (8.40-43.08 ng/ml) were observed 2-24 h after drug administration during the spring treatment and 2-36 h (6.43-24.86 ng/ml) after administration during the autumn treatment. Significantly higher (p<0.05) ivermectin concentrations were found during the first 4 hours after administration in spring in comparison with those determined after the autumn treatment. The administration of the recommended dose of ivermectin resulted in 100% elimination of parasitic eggs from feces in spring and autumn treatment.

Biochemical parameters in Japanese quails Coturnix coturnix japonica infected with coccidia and treated with Toltrazuril.

The activity of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase, total protein, albumin and cholesterol levels were determined in the blood serum of Japanese quails infected with coccidia and treated with Baycox (active ingredient: toltrazuril). Lower levels of AST and ALT activity were noted in treated birds regardless of the applied Baycox dose. The biochemical changes observed in the blood serum of Japanese quails point to coccidia-induced damage of digestive system tissues despite an absence of pronounced clinical symptoms. Significantly lower levels of AST activity and higher levels of LDH activity in treated birds indicate that coccidiosis treatment with toltrazuril contributed to the regeneration of digestive system tissues. An insignificant increase in cholesterol levels was noted, whereas the other serum biochemical parameters remained within the reference ranges.

Toltrazuril (Baycox) treatment against coccidiosis caused by Eimeria sp. in Japanese quails (Coturnix coturnix japonica).

Coccidiosis is the most predominant parasitic disease affecting Japanese quails (Coturnix coturnix japonica) in commercial farms. Coccidiosis as a subclinical infection is difficult to diagnose without parasitological examinations. Oocysts of two Eimeria species, E. bateri and E. tsunodai, were determined in the analysed quail flock. Infected birds were administered Baycox 2.5% at the dose of: group I--7 mg toltrazuril/kg BW per day provided in drinking water (1.5 ml/0.5 1 H2O) that was available 24 h for 2 days, group II--14 mg/kg BW (3 ml/0.5 1 H2O), and group III-- 24.5 mg/kg BW (5 ml/0.5 1 H2O); in groups II and III, the solutions were available 8 h/24 h for 2 days. After the first day of the treatment, the number of excreted oocysts (OPG - oocysts per gram) increased, a steady decrease in oocyst counts began on the second day of Baycox administration and lasted until a three-day period when no oocyst were determined in faecal samples. Regardless of the dose applied, toltrazuril (Baycox) completely eliminated E. bateri coccidia and led to a highly significant reduction in the number of E. tsunodai oocysts. The results suggest that the effectiveness of toltrazuril varies depending on coccidia species and developmental stages of the parasite. From the clinical point of view, the treatment applied significantly reduces the number of coccidia oocysts in commercial flocks of Japanese quails.

Ataxia with isolated vitamin E deficiency is caused by mutations in the alpha-tocopherol transfer protein.

Ataxia with isolated vitamin E deficiency (AVED) is an autosomal recessive neurodegenerative disease which maps to chromosome 8q13. AVED patients have an impaired ability to incorporate alpha-tocopherol into lipoproteins secreted by the liver, a function putatively attributable to the alpha-tocopherol transfer protein (alpha-TTP). Here we report the identification of three frame-shift mutations in the alpha TTP gene. A 744delA mutation accounts for 68% of the mutant alleles in the 17 families analysed and appears to have spread in North Africa and Italy. This mutation correlates with a severe phenotype but alters only the C-terminal tenth of the protein. Two other mutations were found in single families. The finding of alpha TTP gene mutations in AVED patients substantiates the therapeutic role of vitamin E as a protective agent against neurological damage in this disease.

Treatment of cyathostominosis with ivermectin and its influence on selected blood biochemical parameters.

Infections caused by nematodes of the subfamily Cyathostominae affect nearly 100% of pastured horses. Despite of an absence of pronounced symptoms, cyathostominosis can have very serious health consequences. The aim of this study was to monitor changes in total protein levels and concentrations of selected microelements and macroelements in the blood of horses before and after ivermectin treatment. In healthy horses infected by the studied parasites, total blood protein levels were below the physiological norm, but iron (Fe), magnesium (Mg), calcium (Ca) and phosphorus (P) concentrations were within normal limits. Ivermectin treatment reduced the number of excreted parasite eggs (FEC) by 100%, and dead parasites were observed in feces. Decreased iron (Fe) concentrations and an insignificant increase in total blood protein levels were reported. A progressive decline in iron levels was observed when parasite eggs reappeared in feces 60 days after treatment. Iron loss takes place as a result of bleeding from the large intestine when adult nematodes affected by the drug are removed from intestine and fourth-stage larvae leave parasitic nodules in the intestinal wall. A drop in iron levels could be an indirect indicator of the severity of cyathostominosis.

Modulation of specific biochemical blood parameters by helminth infection in laboratory Beagle dogs.

The objective of this study was to examine the independent effect of helminths infection on biochemical blood parameters in Beagles intended for laboratory use which may contribute to a change of experimental results. As a result of research, the authors confirmed the negative effect of helminth invasion on the metabolism of the liver and kidney in laboratory dogs. Stool samples from thirty Beagle puppies were examined for parasites before the puppies were moved to the animal facility, and all were dewormed with Vetminth paste on the day they were moved. Stool examination was performed three more times and animals were given Drontal Plus flavor (Bayer) and Baycox 5% (Bayer). A fourth parasitological examination revealed no intestinal parasites in the feces. Three blood biochemical tests were performed. Experimental results clearly indicate the significant impact of intestinal parasites in dogs used in experiments.

"Virtual patch clamp analysis" for predicting the functional significance of pathogenic variants in sodium channels.

OBJECTIVE: Opening of voltage-gated sodium channels is crucial for neuronal depolarization. Proper channel opening and influx of Na+ through the ion pore, is dependent upon binding of Na+ ion to a specific amino-acid motif (DEKA) within the pore. In this study we used molecular dynamic simulations, an advanced bioinformatic tool, to research the dysfunction caused by pathogenic variants in SCN1a, SCN2a and SCN8a genes. METHOD: Molecular dynamic simulations were performed in six patients: three patients with Dravet syndrome (p.Gly177Ala,p.Ser259Arg and p.Met1267Ile, SCN1a), two patients with early onset drug resistant epilepsy(p.Ala263Val, SCN2a and p.Ile251Arg, SCN8a), and a patient with autism (p.Thr155Ala, SCN2a). After predicting the 3D-structure of mutated proteins by homology modeling, time dependent molecular dynamic simulations were performed, using the Schrödinger algorithm. The opening of the sodium channel, including the detachment of the sodium ion to the DEKA motif and pore diameter were assessed. Results were compared to the existent patch clamp analysis in four patients, and consistency with clinical phenotype was noted. RESULTS: The Na+ ion remained attached to DEKA filter longer when compared to wild type in the p.Gly177Ala, p.Ser259Arg,SCN1a, and p.Thr155Ala, SCN2a variants, consistent with loss-of-function. In contrast, it detached quicker from DEKA than wild type in the p.Ala263Val,SCN2a variant, consistent with gain-of-function. In the p.Met1267Ile,SCN1a variant, detachment from DEKA was quicker, but pore diameter decreased, suggesting partial loss-of-function. In the p.Leu251Arg,SCN8a variant, the pore remained opened longer when compared to wild type, consistent with a gain-of-function. The molecular dynamic simulation results were consistent with the existing patch-clamp analysis studies, as well as the clinical phenotype. SIGNIFICANCE: Molecular dynamic simulation can be useful in predicting pathogenicity of variants and the disease phenotype, and selecting targeted treatment based on channel dysfunction. Further development of these bioinformatic tools may lead to "virtual patch-clamp analysis".

No association of vitamin D intake or 25-hydroxyvitamin D levels in childhood with risk of islet autoimmunity and type 1 diabetes: the Diabetes Autoimmunity Study in the Young (DAISY).

AIMS/HYPOTHESIS: The aim of the study was to investigate the association between vitamin D intake and status and the risk of islet autoimmunity (IA) and subsequent type 1 diabetes in children at increased risk of type 1 diabetes. METHODS: The Diabetes Autoimmunity Study in the Young (DAISY) in Denver, CO, USA, has been following children at increased risk of diabetes since 1993. As of February 2011, 198 children developed IA during follow-up of 2,644 DAISY children. Vitamin D intake and plasma 25-hydroxyvitamin D [25(OH)D] were measured longitudinally. Proportional hazards regression analyses of time to IA, or type 1 diabetes in IA-positive children, were conducted, with vitamin D intake and 25(OH)D as time-varying covariates. HRs were calculated for a standard deviation difference in exposure, with adjustment for confounders. RESULTS: Intake of vitamin D was not associated with the risk of IA (adjusted HR 1.13; 95% CI 0.95, 1.35; p = 0.18) nor progression to diabetes in IA-positive children (adjusted HR 1.30; 95% CI 0.91, 1.86; p = 0.15). Moreover, 25(OH)D level was not associated with the risk of IA (adjusted HR 1.12; 95% CI 0.88, 1.43; p = 0.36), nor progression to diabetes in IA-positive children (adjusted HR 0.91; 95% CI 0.68, 1.22; p = 0.54). In the 128 children in whom we measured 25(OH)D at 9 months of age, 25(OH)D was not associated with risk of IA (n = 30 IA-positive children) (adjusted HR 1.02; 95% CI 0.96, 1.07; p = 0.58). CONCLUSIONS/INTERPRETATION: Neither vitamin D intake nor 25(OH)D levels throughout childhood were associated with the risk of IA or progression to type 1 diabetes in our population.

Cardiopulmonary manifestations of portovenous shunts from congenital absence of the portal vein: pulmonary hypertension and pulmonary vascular dilatation.

HPS and PPHTN are unusual and challenging pulmonary manifestations of liver disease. We report two pediatric cases in association with heterotaxy polysplenia syndrome and congenital absence of the portal vein. Both patients were symptomatic and hemodynamically compromised and required aggressive medical therapy. One patient with PPHTN alone achieved a successful liver transplant. The second child presented with combined HPS and PPHTN and exhibited a different evolution of pulmonary vascular disease. These cases illustrate associations that must be entertained in the setting of heterotaxy syndrome, cyanosis, or pulmonary hypertension and how strategic medical combined with surgical management can provide a good outcome.

A comparison of the effectiveness of the microscopic method and the multiplex PCR method in identifying and discriminating the species of Nosema spp. spores in worker bees (Apis mellifera) from winter hive debris.

The objective of this study was to compare the effectiveness of the multiplex PCR method and traditional light microscopy in identifying and discriminating the species of Nosema spp. spores in worker bees from winter hive debris in the Province of Warmia and Mazury (NE Poland). A total of 1000 beesdead after from the bottom of the hive from bee colonies were analyzed. Spores were identified with the use of a light microscope (400-600x magnification). Spores were assigned to species by the multiplex PCR method. The microscopic evaluation revealed the presence of Nosema spp. spores in 803 samples (80.3%). Nosema ceranae spores were observed in 353 positive samples (43.96%), Nosema apis spores were found in 300 samples (37.35%), while 150 samples (19.67%) showed signs of a mixed infection. A multiplex PCR analysis revealed that 806 samples were infested with Nosema spp., of which 206 were affected only by Nosema ceranae, 600 showed signs of mixed invasion, while no samples were infected solely by Nosema apis parasites. In two cases, the presence of spores detected under a light microscope was not confirmed by the PCR analysis. The results of the study indicate that Nosema ceranae is the predominant parasitic species found in post-winter worker bees from the bottom of the hive in the region of Warmia and Mazury.

Histopathological changes of the skin in hens infested with Dermanyssus gallinae.

Studies were carried out on dead hens from a commercial farm of layer hens. Sections of skin from the region of the breast and under the wings with changes following mite bites were taken for histopathological examinations. In the histopathological preparations hyperkeratosis of epidermis and pachydermatosis were ascertained. Numerous lymphocytic cells focally infiltrated the connective tissue and were present under the epidermis. Subcutaneous connective tissue was oedematous. Furthermore, excessive desquamation of the corneal layer of the epidermis and small epidermoidal cysts in dermis occurred, containing callous cells and amorphous protein substances.

Changes in the corticosterone level in tooting male black grouse (Tetrao tetrix) infected with Eimeria spp.

The black grouse (Tetrao tetrix) is a forest bird species critically endangered of extinction. Enclosed aviary breeding is among the measures taken to protect the species. Complex factors can affect its reproduction success, including coccidiosis. In this article, corticosterone level (as a reliable biomarker of stress in birds) and prevalence of Eimeria spp. were determined in male black grouse kept in aviary breeding center before, during, and after the reproductive season (called tooting). The correlation between those parameters was also analyzed. The corticosterone level was measured with noninvasive method in fecal samples in an immunoassay. The prevalence of Eimeria spp. was determined by the qualitative (Fülleborn's flotation) and quantitative method (McMaster's method as modified by Raynaud). Results show the occurrence of 3 species of Eimeria spp. in males: Eimeria lyruri, Eimeria nadsoni, and Eimeria nonbrumpti. Infestation with E. lyruri was chronic in nature. A co-invasion of E. lyruri, E. nadsoni, and E. nonbrumpti was observed during (prevalence 60%) and after the tooting (prevalence 40%). The study showed no statistical changes in the oocysts per 1 g of feces (CORTl) and oocysts per 1 g of feces (OPG) in assays I-III and absence of correlation between CORTl and OPG. The results of this experiment do not support the hypothesis that chronic infestation with Eimeria spp. can induce chronic stress in grouse. But may suggest that males of black grouse are susceptible to infestation with other species of Eimeria spp. during mating season. The knowledge of the level of individual stress and parasitic infestation can be used to take protective actions for this bird species, especially to achieve higher survival rate and bird reproduction rate. Tangible effects will include an assessment of the relationship between individual susceptibility and chronic environmental stress caused by coccidia.

Influence of Dermanyssus gallinae (poultry red mite) invasion on the plasma levels of corticosterone, catecholamines and proteins in layer hens.

The results of studies conducted in 2006 revealed that mass red mite (Dermanyssus gallinae) invasions cause somatic stress which may be responsible for the pathophysiological mechanism of decreased egg production, lower humoral immunity and higher mortality in layer hens. The aim of this study was to validate the above research results, to investigate whether in addition to somatic stress, red mite invasions cause psychogenic stress due to the activation of the sympatho-adrenomedullar system, and to determine the level of stress resulting from red mite infestations in comparison with a short, 1.5 h period of acute immobilisation stress. The study investigated 36 HY-Line Brown layer hens divided into three groups: a non-infested control group, an experimental group infested with red mites and a non-infested experimental group subjected to acute immobilisation stress for 1.5 h. Blood samples were taken from all hens for the determination of the levels of corticosterone, adrenaline, noradrenaline, albumin, and alpha-, beta- and gamma-globulins. The results validated the previous reports on the occurrence of somatic stress and on a significant decrease in y-globulin levels (p < or = 0.01) in the group of birds infested with red mites, in comparison with the control group. Adrenaline levels in infested hens were indicative of psychogenic stress. Based on a comparison of hormonal indicators in all hen groups, the level of somatic stress resulting from red mite infestation can be classified as moderate, while the level of psychogenic stress can be interpreted as high. A significant drop in y-globulin levels in the blood of birds infested with red mites also shows that the invasion induces chronic stress which lowers the humoral immunity of hens.

The influence of administering "effective microorganisms" to pullets on chosen haematological and biochemical blood indexes.

"Effective Microorganisms" (EM)--a mixture of lactic acid bacteria, photosynthetic bacteria, yeasts and fungi are used mainly in agriculture and organic waste treatment. Recently, they have also been added to water and feed for animals, as well as to processing their excrements into compost and to eliminate the stench. The objective of the present study was to assess the influence of a 14-day administration of an EM solution in drinking water to layer hens on chosen haematological and biochemical indexes. The research was carried out on 120 hens divided into two equal groups. The birds in the experimental group were given drinking water with dissolved EM (5% solution), and those in the control group--water without the preparation. On the 64th day of the aviculture, the hens were weighted and their blood was taken from the wing vein for haematological and biochemical examinations. Administering EM with water to hens did not influence significantly their body weight nor chosen haematological and biochemical indexes. A significant increase was found only in the number of platelets, the level of albumins, the content of total cholesterol and the LDH activity, however, a decrease in the ALT activity was observed.

European biliary atresia registries: summary of a symposium.

Biliary atresia (BA) is a rare but potentially devastating disease. The European Biliary Atresia Registry (EBAR) was set up to improve data collection and to develop a pan-national and interdisciplinary strategy to improve clinical outcomes. From 2001 to 2005, 100 centers from 22 countries registered with EBAR via its website (www.biliary-atresia.com). In June 2006, the first meeting was held to evaluate results and launch further initiatives. During a 5-year period, 60 centers from 19 European countries and Israel sent completed registration forms for a total of 514 BA patients. Assuming the estimated incidence of BA in Europe is 1:18,000 live births, 35% of the expected 1488 patients from all EBAR participating countries were captured, suggesting that reporting arrangements need improvement. At the meeting, the cumulative evaluation of 928 BA patients including patients from other registries with variable follow-up revealed an overall survival of 78% (range from 41% to 92%), of whom 342 patients (37%) have had liver transplants. Survival with native liver ranged from 14% to 75%. There was a marked variance in reported management and outcome by country (e.g., referral patterns, timing of surgery, centralization of surgery). In conclusion, EBAR represents the first attempt at an overall evaluation of the outcome of BA from a pan-European perspective. The natural history and outcome of biliary atresia is of considerable relevance to a European population. It is essential that there is further support for a pan-European registry with coordination of clinical standards, further participation of parent support groups, and implementation of online data entry and multidisciplinary clinical and basic research projects.

Influence of light and darkness on the behaviour of Dermanyssus gallinae on layer farms.

The behaviour of Dermanyssus gallinae was investigated on two layer farms where two different light programs were introduced in the 40th week of hen life. In layer house No. 1, light was applied continuously for 16 hours during the day, while layer house No. 2 was subjected to 4 hours of light and 2 hours of darkness applied alternately during the day. To monitor the level of red mite infestation, 30 tube traps were placed in every layer house corridor at a height of 1.5 m above the floor. In the first layer house, 280 Dermanyssus gallinae females, 50 nymph larvae and 198 eggs were found in 100 mg of tube trap material during 16 hours of the light phase, while during the 8-hour darkness phase, 1240 females, 70 nymph larvae and 110 eggs were collected. In the other layer house (with an alternating light phase of 4 hours and a darkness phase of 2 hours per day), 387 Dermanyssus gallinae females, 401 nymph larvae and 1060 eggs were found in trap tubes over the 8-hour dark phase, while 343 females, 202 nymph larvae and 1106 eggs were discovered over the 16-hour light phase.

Measuring beverage consumption in US children and adolescents: a systematic review.

Beverage consumption habits are associated with weight outcomes for children and adolescents. Many studies have examined youth's beverage consumption, but little is known about what methods are commonly used to assess youth beverage consumption and whether these strategies are valid and reliable. This study aimed to systematically review articles assessing beverage consumption among children and adolescents. We searched PubMed and Scopus for English-language articles published between February 2007 and February 2017 that measured and reported on American youth's (ages 2-18 years) beverage consumption. Searches yielded 17,165 articles, of which 589 articles describing 615 measures were extracted. We examined the types of assessment methods used, characteristics of these methods (e.g. validity, reliability, and literacy level), characteristics of study samples, and beverages assessed. The most common assessment methods were questionnaires/screeners (used by 65.4% of articles) and recalls (24.4%). About three-quarters of articles did not address validity (70.5%) or reliability (79.5%) of any measures used. Study populations were diverse: 54.7% of articles included low-income children, and 90.2% included non-White children. The most commonly assessed beverage category was sugar-sweetened beverages. Findings suggest that improved measurement techniques and reporting are both needed to track progress towards a goal of ensuring all youth have healthy beverage consumption.