Volume 56 (2012): Issue 1 (March 2012)

The liver and kidneys of broilers fed during the first two weeks of fattening a fodder containing the addition of 0.025% propolis and/or 0.5% pollen were examined macroscopically and microscopically. Additionally, the effects of these substances on the results of breeding and on pathomorphology of the liver and kidneys in the course of natural, asymptomatic infection with Salmonella Enteritidis were determined. The results demonstrated that propolis has protective effects on the liver of broiler chickens, reducing the intensity of regressive lesions. However, regarding supplementation of the diet with bee pollen, additional research on its effects in animals is needed.

The objective of this study was to determine whether insertion-deletion (indel) 23 bp polymorphism of the bovine prion protein (PRNP) gene differentiates the total number of leukocytes and lymphocytes and the number of virus-infected lymphocytes. The experimental materials comprised 119 Black-and-White Polish Holstein-Friesian cows. Bovine leukosis was diagnosed by an indirect immunofluorescence based on the detection of viral protein p24 in bovine lymphocytes infected with leukaemia virus (BLV). Indel23 polymorphism was determined by PCR. Blood haematological parameters (total leukocyte counts, total lymphocyte counts, and their percentages) were determined at a specialist haematological laboratory. The examined indices were analysed in three replications, at one-month intervals. It was found that indel23 polymorphism significantly differentiated blood leukocyte counts and the total number and percentage of lymphocytes. Cows with the 23 bp del/del genotype showed significantly higher leukocyte and lymphocyte counts than animals with the remaining two genotypes. Higher values of the analysed haematological parameters noted in homozygotes with 23 bp deletion are similar to the values reported in cows affected by persistent lymphocytosis, thus pointing to an adverse effect of this genotype on the haemopoiesis process. The variations between indel23 genotypes and the number and percentage of BLV-infected lymphocytes are less obvious and more difficult to interpret.

The aim of this study was to describe the state of infection of roe deer (Capreolus capreolus) and red deer (Cervus elaphus) by Bartonella sp. in North-Western Poland through PCR detection of Bartonella 16S-23S rRNA ITS region in isolates of animal tissues, and also to describe the genetic diversity of detected Bartonella species based on molecular analysis of ITS. The multiple alignment analysis of ITS sequences was carried out, and homology matrices and phylogenetic trees were constructed. The DNA of Bartonella sp. was detected in tissues of 45.6% (36/79) C. capreolus and of 50% (15/30) C. elaphus. Products of two different sizes were detected: 317 bp, characteristic for B. schoenbuchensis, and 198 bp, characteristic for B. bovis. The obtained results suggest that roe and red deer are potential reservoirs of Bartonella sp. Most of the analysed ITS sequences was not specific for one host species. In constructed phylogenetic trees, sequences obtained from roe and red deer clustered together. These results suggest a lack of host specificity of most detected B. schoenbuchensis and B. bovis intraspecies strains

The presented investigations were conducted on a group of 60 porkers of crossbreed Polish Landrace x Large White Polish. The animals were divided into two equal experimental groups. The control group (K) was fed diets without supplementation with probiotics, group (P) - diets with the addition of probiotic (0.2 kg t-1 feed). The aim of the study was to determine the effect of probiotic preparation on total numberof lactic acid rods from the Lactobacillus genus and those forming hydrogen oxide. The second part of experiment concerned the influence of probiotic preparation on the number, haemolytic ability and changes in drug resistance of Escherichia coli isolated from animal faeces. The significantly highest number of Lactobacillus sp. were determined in the saliva of porkers fed diets with the addition of probiotic, while the lowest in the control group. Lactobacillus sp. rods capable of forming hydrogen peroxide were isolated from 17 animals in group K and from three animals in group P. E. coli was determined in each examined sample of faeces. In groups K and P, counts of these bacteria were similar and did not differ statistically. High numbers of haemolytic isolates (haemolysis β) were found in faeces of animals fed diets with the addition of probiotic. Number and proportions of resistant isolates in groups K and P were different. Gentamicin was characterised by exceptionally high in vitro effectiveness. The used probiotic increased drug resistance of E. coli and increased frequency of incidence of haemolysis β.

The purpose of this study was to present the structure of nematofauna community with particular emphasis on stomach (proventriculus and gizzard) nematodes in wild ducks wintering in large numbers in the North-Western Poland. Hosts (n=152) were represented by 17 species from eight genera belonging to three different ecological tribes: Anatini, Aythyini, and Mergini. Parasitic nematodes (n=14,396) were found in the digestive tract of 813 out of 1,052 birds (77.3%), with the vast majority of nematodes (93.9%) found in the gizzard (n=7,326) and proventriculus (n=6,198). Nematodes isolated from these organs represented 21 species from six families: Amidostomatidae, Acuariidae, Tetrameridae, Dioctophymatidae, Ascarididae, and Anisakidae. The most prevalent were nematodes from the genus Amidostomoides (n=6,686 individuals; 49.4%), and the most dominant species was A. monodon (5,013 nematodes).

The aim of this study was to determine the prevalence of infection with gastrointestinal parasites in water buffaloes raised in the Notecka Forest region in the Wielkopolska Province (Poland). Coproscopic examinations were performed between January and June 2009. The mean prevalence of gastrointestinal parasites in the water buffalo herd was 44%. Gastrointestinal nematodes (10%) and flukes (Fasciola hepatica - 32% and Paramphistomum cervi - 11%) were found in the examined animals. The high prevalence of infection with gastrointestinal parasites was caused by the lack of an appropriate helminth control programme. Despite the fact that they are raised on marshy meadows, where good conditions for parasites’ development exist, these animals were not systematically dewormed. It is necessary to devise a helminth control programme for the studied water buffalo herd.

The aim of the study was cloning and analysis of the entire coding sequence of hamster IL-2 by the method of RACE-PCR, its expression in Escherichia coli cells, and production of IL-2 specific antibodies. These antibodies were used to determine in vitro IL-2 production by cells derived from the spleen and mesenteric lymph nodes of Ancylostoma ceylanicum infected hamsters. The highest concentration of IL-2 was noted in supernatants from cell cultures coming from the oldest, most resistant hamsters.

The research aimed at evaluating the epizootic and epidemiological situation of trichinosis during the last 8 years in Lublin province on a background of progressing increase in wild boar population within the region and in the whole country. Data for the study were taken from the report on the results of the official examination of slaughter animals and meat, poultry, game, lagomorphs and aquaculture animals and six reports on the number of trichinosis cases found at wild boars and domestic swine. In order to evaluate the trichinosis epidemiological situation within the region, reports of the National Institute of Public Health-National Institute of Hygiene on the number of identified trichinosis cases in people, as well as the number of hospitalised patients were presented. In addition, information on the population and hunting achievement of wild boars in hunting circuits of Lublin province during the last 8 years was enclosed. The number of identified trichinosis cases in meat of wild boars from Lublin region increased 9 times, while the percentage of trichinosis occurrence in reference to the number of examined carcasses almost 3-fold. At the same period, the number of porcine carcasses, in which trichinosis was found, decreased by over 4 times. Over double increase in wild boar population on the studied area was observed during the evaluation. Dynamic increase in the population size - in an aspect of the species population interaction with the living habitat, and in the form of the increase in the number of damages of crops and cultivation fields - contributed to intensified hunting pressure towards the species expressed as almost 3-fold increase of wild boar hunting. Analysis of epizootic and epidemiological situation of Poland indicates that wild boar meat was the principal source of trichinosis during the studied period. Considering Lublin province, the number of identified trichinosis cases is still high as compared to eastern and central provinces. Meanwhile, when compared to western and northern Poland, the level of trichinosis invasion can be considered as low. Furthermore, the trichinosis morbidity among people, that does not exceed 0.18/100 thousand inhabitants, can be regarded as low. Nevertheless, the fact of underestimating the necessity of both wild boar’s and swine’s meat examination seems to be alarming

The objective of the presented study was to determine microbial contamination of calf muscle tissues in relation to slaughtering process of calves during a slaughter day. In order to determine the total aerobic bacteria count, and the presence of bacteria from the Enterobacteriaceae family and enterococci, 32 carcasses were examined (eight each slaughter day), while 40 carcasses (10 each slaughter day) were examined for the detection of Salmonella organisms. Microbiological evaluation of each bacterial group was performed according to the Polish Norms. In most cases, no significant differences were reported between the total aerobic counts on calf carcass surfaces as determined at each slaughter cycle. The daily log mean values were lower by 1 up to 1.5 log, respectively, than the maximal bacteria count (M). Bacteria from the Enterobacteriaceae family were isolated from 59.4% of the evaluated samples. However, contamination with these bacteria was insignificant - 1.7 x 10 cfu/cm². Enterococci were isolated from all investigated samples and contamination ranged from 1.1 x 10 cfu/cm² up to 4.1 x 102 cfu/cm². Salmonella strains were not found in any of the evaluated samples. The order of the slaughtering calves during a slaughter day and the day of the week when the examination was performed did not have any influence on total microbial contamination of carcasses. Thus, sanitary conditions in the examined abattoir were satisfactory and slaughter process was conducted at a high quality level

The aim of the study was to evaluate the oxygen metabolism of neutrophils in peripheral blood of mares in relation to intensity of endometrium inflammations. The study involved 36 half-breed mares, aged 4-22 years, showing fertility disturbances. In 26 mares neutrophils were found in uteral smears, which indicated endometritis (15 - moderate inflammation and 11 - severe inflammation). In the rest mares, cytological examination excluded inflammation. Blood samples were evaluated in terms of neutrophils chemiluminescence without stimulation (CL-WS) and with stimulation by opsonised zymosan (CL-OZ). The study demonstrated (only in case of CL-WS) an increase in chemiluminescence of cells in mares with a severe inflammation of the endometrium. The increased chemiluminescence activity was accompanied by a decrease in activation index (OZ/WS) of neutrophils, suggesting some imbalance between production and elimination of reactive oxygen species (ROS). The correlation analysis demonstrated a statistically significant relation between the intensity of the uterus inflammation, which was verified by cytological examination and CL-WS of peripheral blood neutrophils, as well as their activation index. The obtained results suggest that activated neutrophils are an important source of ROS which can play a role in the pathogenesis of endometritis in mares.

In in vitro study on green monkey kidney (GMK) cell culture, the cytotoxicity of Ukrain, methotrexate (MTX), and their combination was investigated. The effect of these drugs and their combination on viability (MTT test) and apoptosis of the cells was assessed. The IC₁₀ and IC₅₀ values for Ukrain and MTX were also indicated. After 24 h of incubation of GMK cells with Ukrain, IC10 amounted to 84.6 μmol/L and IC₅₀ was 256.3 μmol/L, while MTX IC₁₀ amounted to 7.18 μmol/L and IC₅₀ was 154.8 μmol/l. After 24 h of simultaneous incubation of GMK cells with 50 μmol/L of Ukrain and 5.5 μmol/L of MTX, 15.33 % of cytotoxicity of the drugs in LDH test was found. The most significant increase in the cytotoxicity (42.10 %) was noted after 24 h incubation of GMK cells with 150 μmol/L of Ukrain and 5.5 μmol/L of MTX. Likewise, in the MTT assay the greatest decrease in the cells viability was found after incubation with 150 μmol/L of Ukrain and 5.5 μmol/L of MTX. The evaluation of apoptosis also indicated the adverse effect of combined application of both drugs on GMK cells.

The presented study was planned to determine whether vasoactive intestinal peptide (VIP) could prevent cytokine haemostatic, haematological, and biochemical disturbances in LPS-treated rats. Adult male Wistar rats (weight range: 200-250 g) were used. The study included four groups: group 1 served as a control group (C); animals in group 2 were given intravenously 1.6 mg/100 g of LPS (E. coli, serotype 0.111:B4); in group 3, rats were injected intraperitoneally with 25 ng/kg of VIP; in group 4, the same doses of VIP and LPS were injected simultaneously. Blood samples were collected 6 h after treatments. In endotoxaemic rats, platelet count, fibrinogen, and antithrombin levels were decreased, the activated partial thromboplastin time and prothrombin time were prolonged, and leucopoenia, as well as significant changes in differential leukocyte percentage were demonstrated. In addition, LPS caused statistically significant increases in plasma TNF- , IL-6, and IL-10 levels, and AST, ALT, creatinine, cholesterol, triglyceride concentrations. However, it caused a statistically significant decrease in total protein and albumin levels when compared to control group. The results showed that during endotoxaemia, VIP had moderately therapeutic effect as an antiinflammatory agent, suppressing TNF-α and IL-6, and stimulating IL-10; however, it was not effective against the adverse effect of LPS on investigated haematological and biochemical parameters.

Results of supravital and post-mortem observation of embolic-thrombotic complications in dogs, in the course of endotoxaemia induced by pyometra and phlegmonous pneumonia due to infection with Pseudomons aeruginosa, or endomyocarditis caused by Staphylococcus aureus infection were described. Multiorgan embolic-thrombotic complications were diagnosed in all presented cases. Most probably, the complications developed due to thrombocytopoenia secondary to platelet activation and exacerbation of endotoxin-induced DIC syndrome. In all the dogs, a disturbed cardiac rhythm was diagnosed supravitally. The rhythm disturbances might reflect myocardial injury due to presence of bacterial endotoxins in blood. The myocardial injury was confirmed by histopathological examination.

The clinical form of natural zearalenone intoxication was observed in sheep that were kept indoors and fed a constant diet of feed concentrates containing high concentrations of zearalenone and its metabolite α-zearalenol. The clinical form of the disease was not noted in the control group, consisting of sheep that were kept on a pasture from spring to late autumn; only in the winter they were fed wheat pellets, in which the zearalenone concentration was determined to be the lowest among all used feed concentrates. During the course of natural zearalenone intoxication, metabolism of arachidonic acid increased, mainly due to enzymes of the cyclooxygenase group, which are responsible for the generation of prostaglandin F2α and thromboxane B2, and an increase in their concentration. Increased production of F2α and B2 was closely correlated with the serum level of SAA, an indicator of the intensity of the inflammatory reaction. This indicates that both compounds participated in the development of inflammatory reactions in the terminal end of the digestive tract that accompanied zearalenone intoxication in sheep. The imbalances noted between the eicosanoid classes investigated in this study were fundamentally responsible for the development of clinical symptoms in sick sheep that showed symptoms of partial or total prolapse of the anus and rectum and prolapse of the large intestine, which were the direct cause of the animals death.

Expression of α (ER-α) and β (ER-ß) oestrogen receptors in neurons of basolateral complex of the amygdala was studied in rabbit males. The examinations were carried out on brains of 12 sexually mature rabbit males weighing 3-4 kg. An immunohistochemical method (IHC) with primary Er-α antibodies, NCL-L6F11 and Er-ß: NCL-ERbeta clone EMR02 was applied. The percentage of immunoreactive neurons was determined. Morphometric and statistical analysis of immunoreactive (ER+) neurons was carried out including the localisation of IHC reaction within the examined neurons. The main place of IHC-ER-α reaction was the cytoplasm of nervous cells, whereas in the examination of IHC-ER-β, the reaction was observed mainly within cell nucleus. The nuclei of basolateral complex are a region of relatively high percentage of ER-β+ neurons and lower percentage of ER-α+ neurons in comparison with other nuclei of the amygdala. Basolateral nucleus was characterised by a higher percentage of ER- β+ neurons in which the reaction was observed in the cytoplasm.

The aim of this study was immunohistochemical evaluation of heat shock protein (Hsp70) and p-53 proteins in the Larginine- induced cellular stress in hepatocytes of rats. Sixteen white Wistar female rats were divided into two equal groups. The rats from the experimental group received per os 40 mg/kg b.w. of L-arginine every day for 2 weeks and were decapitated after 3 weeks of the experiment. The rats from the control group received in the same manner 2 ml of distilled water and were decapitated after 3 weeks of the experiment. After decapitation specimens from the liver were collected, fixed in 10% formalin, and then embedded in paraffin blocks. Proteins Hsp70 and p-53 were detected on slides using the standard three step immunohistochemical method. The quantitative evaluation of Hsp70 and p-53 expression showed that the area of positive staining in the liver of the experimental rats (Hsp70 305,763.00 μm² +/-58,289.66, p-53 9,551.42 μm² +/-1,078.86) was comparable to that in the control groups (Hsp70 291,636.80 μm² +/-34,492.31, p-53 14,104.67 μm² +/-3,571.35). Our experiment showed, that L-arginine as a precursor of exogenous nitric oxide given to rats in dose similar to that used in pregnant women treated for hypertension did not exhibit an influence on hepatocytes.

The objective of this study was to evaluate the effect of dietary conjugated linoleic acid (CLA) and false flax (Camelinasativa) seed oil (CS) on bone quality in broiler chickens. Experiment was carried out on 96 chickens randomly divided into four groups: control group fed diet with sunflower oil (SO) and three experimental groups fed diet with addition of CLA (2.86% starter, 4.32% grower) and diet with addition of CLA (1.43% starter, 2.16% grower) and SO or CS (1.43% starter, 2.16% grower). On the 35th d of life eight birds per treatment were slaughtered and the femur, humerus, and tibia were isolated for further analysis. Using DXA method, bone mineral content (BMC) and bone mineral density (BMD) were measured. Furthermore, weight and length of bones were assessed. The weight, BMD, and BMC in chickens from CS+CLA group were significantly higher when compared with CLA group. The significant differences were noted for BMC between control and CS+CLA birds. No marked differences of bone parameters were observed between control group and CLA, and SO+CLA groups. In conclusion, the results indicate that false flax oil and CLA can replace sunflower oil in chickens feeding, and demonstrate the effectiveness of false flax oil on the enhancement of bone properties in broiler chickens.

The aim of this study was to demonstrate the diagnostic accuracy of P-wave dispersion in predicting recurrence of atrial fibrillation in dogs with sinus rhythm restoration after external cardioversion. The study was performed on 15 dogs, which underwent electrical external cardioversion after atrial fibrillation. Nine-lead surface electrocardiogram of each dog was recorded 1 min after the cardioversion process to measure the P-wave duration. Dogs with recurrent atrial fibrillation had significantly higher P-wave dispersion compared to dogs that had a sinus rhythm lasting more than 3 months (9.26 ±2.01 ms vs 20.9 ±2.18 ms, P=0.001). This study suggests that P-wave dispersion analysis in dogs after a successful external electrical cardioversion has diagnostic accuracy to predict the recurrence of atrial fibrillation.

The aim of the study was to define morphology and distribution of calretinin (CR) positive neurons in the frontal cortex of adult chinchilla males and intracellular localisation of the protein in this area. The brains of 5 adult chinchilla males were used in the study. CR immunoreactive neurons were shown with peroxidase-antiperoxidase immunohistochemical reaction using a specific monoclonal antibody. Intensive CR immunoreactivity was demonstrated mainly in few polymorphic neurons of II, III, and V layers. Cytoplasmic and nuclear reaction product in most CR positive neurons was diffuse and in some neurons of layer V in the form of granules localised peripherally. In a few cells more intensive staining was observed in the nuclei than in the cytoplasm. The results indicate the presence of heteromorphic CR positive neurons in specific layers of the frontal cortex. Nuclear localisation of CR in neurons suggests passive transport of this protein, which may affect the nuclear genes. This protein is a neuroprotector maintaining appropriate level of calcium, modulating neuronal activity, and synaptic conduction in the frontal cortex of chinchilla.

The aim of the study was to analyse the effects of maternal obesity and the litter size on the growth rate and plasma concentrations of GH, IGF-1, insulin, and glucose in ewe lambs during the first 6 weeks of their postnatal life. Seventy-six SCP sheep: 35 ewes and 41 female offspring were used. Before gestation, the ewes were divided into two groups: N - normally weighing and O - obese sheep. After the parturition, the lambing rate and the birth type were estimated. The born female lambs were separated into five groups: IN - singleton and IIN - twin offspring of normally weighing mothers; IO - singleton, IIO - twin and IIIO - triplet offspring of obese mothers. They were weighed at birth and weekly thereafter, until the completion of the 6th week of their postnatal life. Afterward, daily weight gains and concentrations of biochemical parameters were analysed. No significant differences in GH, IGF-1, insulin, and glucose levels between the groups of lambs born to non-obese and obese sheep, both carrying singletons and twins, were found. In contrast, significantly increased concentrations of GH (P≤0.001), insulin (P≤0.001), and glucose (P≤0.05) and markedly dropped level of IGF-1 (P≤0.001), as well as reduced daily body mass gains in triplets in comparison to other groups of lambs were observed. Maternal obesity caused significant lambing rate’s accretion with the rise in triplets’ frequency. However, in ewe lambs of this birth type, the disrupted relationships between plasma levels of GH, IGF-1 and growth rate, and between plasma levels of insulin and glucose were found.