The effect of feed supplementation with a copper-glycine chelate and copper sulphate on selected humoral and cell-mediated immune parameters, plasma superoxide dismutase activity, ceruloplasmin and cytokine concentration in broiler chickens.

The varied bioavailability and different effects of organic forms of copper on the immune system of poultry have prompted the search for new feed additives based on copper compounds containing modified chelate complexes. The aim of the study was to determine the effect of inorganic and organic forms of copper on selected parameters of the cellular and humoral immune response in broiler chickens by determining the percentages of CD3+ CD4+ , CD3+ CD8+ and CD25+ lymphocytes, cells with MHC Class II expression, and BU-1+ cells, as well as the concentrations of SOD, IL-2, IL-10 and TNF-α in the peripheral blood. The experiments were conducted using 500 one-day-old Ross 308 roosters divided into five groups. Cu was added in inorganic form (CuSO4 ), in inorganic form with the addition of phytase (CuSO4 + F), in organic form in combination with glycine (Cu-Gly) and in organic form in combination with glycine and a phytase supplement (Cu-Gly+F). The results of the study indicate an increase in the percentage of CD3+ CD4+ and CD3+ CD8+ T lymphocytes, CD25+ T cells, and cells expressing MHC class II molecules, and in the concentration of ceruloplasmin, activity of superoxide dismutase and the concentration of IL-2 in the groups that received copper, particularly copper-glycine chelates. Based on the study, we can conclude that supplementation of poultry feed with copper chelates activates mainly the Th1 cellular immune response and the response of peripheral blood T lymphocytes. Furthermore, it promotes secretion of cytokines, which are involved in potentiation and regulation of the immune response in birds.

Effect of feed supplementation with zinc glycine chelate and zinc sulfate on cytokine and immunoglobulin gene expression profiles in chicken intestinal tissue.

The aim of the study was to evaluate the effect of inorganic and organic forms of Zn on the expression of cytokines (IL-2, TNF-α, IFN-γ, IL-12, IL-17, IL-4, IL-10, and TGF-ß) and immunoglobulins (IgA and IgG) in the tissues of the small intestine (jejunum and ileum) of broiler chickens. In the experiment, 90 broiler chickens were divided into 4 experimental groups and a control group, with 18 birds each. The birds received Zn supplements in inorganic form with and without phytase (ZnSO4 and ZnSO4 + F), and in organic form with glycine, with and without phytase (Zn-Gly and Zn-Gly + F). The total rearing period was 42 days. Quantitative real-time (RT)-PCR was used to measure the expression of the cytokines and immunoglobulins. The differences between the results obtained for the control and experimental groups, between the groups receiving ZnSO4 and Zn-Gly, and between groups ZnSO4-F and Zn-Gly-F were analyzed statistically. High relative expression of IL-2 was observed for the chickens in the groups receiving ZnSO4-F, Zn-Gly, and Zn-Gly-F on d 42 in comparison to the control group. High relative expression of TNF-α, IL-12, and IL-17 was noted in the group that received ZnSO4 + F. High expression of IgG, IgA, IL-4, TGF-ß, and IL-10 was noted in the groups of chickens that received feed supplemented with Zn-Gly and Zn-Gly + F chelates on d 42 of the study in comparison to the control group. In conclusion, supplementation with Zn-Gly chelates can ensure Th1 and Th2 balance during the immune response in the gut-associated lymphoid tissue (GALT), and, by increasing IgA and IgG expression, also can stimulate potentiation of the immune response involved in passive protection of the body from infection. In contrast, the use of inorganic forms of Zn, in the form of sulfates, can induce local inflammatory processes in the intestines, which, in the case of long-term supplementation, lead to the development of infections.

Changes in selected subpopulations of lymphocytes in dogs infected with Babesia canis treated with imidocarb.

OBJECTIVE: The purpose of this study was to track changes in selected subpopulations of lymphocytes in the blood of dogs infected with Babesia (B.) canis and treated with imidocarb. MATERIAL AND METHODS: The study included 16 dogs divided into two groups. The first group (n = 6) consisted of healthy control animals. Dogs of the se- cond group (n = 10) were infected with B. canis and after establishment of the diagnosis each animal received a single dose of imido- carb (5 mg/kg). Flow cytometry was used to enumerate several immune cell phenotypes. RESULTS: It was concluded that the invasion of B. canis contributes to the decreased percentage of CD3+, CD4+, CD8+, CD21+ lymphocytes in the blood of infected animals. The decreased level of tested subpopulations of lymphocytes in group 2 persisted for the entire 12-day period of the test. After the administration of imidocarb, each tested lymphocyte fraction in the blood of the dogs with babesiosis increased, but did not reach physiological values. CONCLUSION: The presented results indicate that the resolution of clinical signs associated with babesiosis may be related to the stimulation and intensity of cellular immunity, dependent on the CD4+ T cells profile. After administration of imidocarb, the parasitemia is cleared which allows the recovery of the lymphocyte populations.

The genetic analysis of new Polish strains of European brown hare syndrome virus.

In this paper we describe recently occurring outbreaks of European brown hare syndrome (EBHS) in a captive hare population. The aim of our study was to evaluate the phylogenetic position of detected Polish strains compared to other European strains of EBHSV. Investigations were undertaken in hares from different provinces of Poland. Liver or spleen samples were tested for viral RNA using the RT-nested PCR method and the products were subsequently sequenced. The genetic analysis was based on the fragment of gene encoding viral capsid protein; it revealed a high homology and close relationship between Polish and European EBHSV strains isolated between 2001 and 2011.

Trueperella pyogenes infections in swine: clinical course and pathology.

Trueperella pyogenes is an opportunistic pathogen causing purulent infections in pigs and other animal species. T. pyogenes infections in pigs are local and/or generalized depending on the immune status of the animals, their individual susceptibility and environmental factors. The occurrence of these infections on pig farms causes substantial economic losses in breeding and rearing. In sows from the breeding herd, the disease leads to infertility, embryonic death, abortion, and disorders of the menstrual cycle and lactation. Mastitis is the major cause of losses in piglets. Disorders of the musculoskeletal system, including inflammatory polyarthritis, fractures and degenerative joint disease results in the culling of animals with high breeding value. In other technological groups, multi-organ inflammations and movement disorders dominate, leading to a reduction of the slaughter value and elimination of pigs from breeding. Understanding of the clinical and pathological aspects of T. pyogenes infections in pigs will enable the development of effective methods of combating this disease on pig farms.

Haemorhagic enterotoxemia by Clostridium perfringens type C and type A in silver foxes.

Type C and type A of C. perfringens were detected in the seat of natural infections in silver foxes characterized by symptoms of haemorrhagic enterotoxemia. In all of the dead foxes characteristic changes were noted in the small intestine and parenchymatous organs. The production of alpha and beta toxins by isolated bacteria was confirmed by the bioassay using white mice and by PCR. The results of the drug sensitivity testing showed that isolated strains were highly susceptible to amoxicillin with clavulanic acid, metronidazole, doxycycline and penicillin with streptomycin.

Molecular analysis of a fragment of gene E1B 19K of canine adenovirus 2 (CAV-2) isolated from dogs with symptoms of cough.

The aim of this study was to perform molecular analysis of canine adenovirus 2 (CAV-2) E1B 19K gene fragment isolated from 20 dogs of various breeds (12 males and 8 females aged 1-9 years), with clinical symptoms of upper respiratory tract infections, from the Lubelszczyzna region. Nasal swabs were taken from dogs. DNA of CAV-2 was detected using the PCR method in 16 swabs. All PCR products were sequenced, and the obtained sequences were compared with each other and with the sequence of the E1B 19K gene of the CAV-2 strain from an online database of NCBI GenBank: AC 000003. Based on analysis of the obtained sequences, three polymorphic variants of CAV-2 (No. 1-3) with homology of 78 - 100% were distinguished. The nucleotide and amino acid sequences of the most frequently represented polymorphic variant, No. 1, differed from the sequences of polymorphic variant No. 2 with one substitution. The nucleotide and amino acid sequence of the E1B 19K gene of CAV-2 AC 000003 differed from the analogous sequences of representatives of variant No. 1 with 44 nucleotide and 19 amino acid substitutions. The small number of nucleotide differences in the E1B 19K CAV-2 gene among the examined own isolates, compared with AC 000003, suggest that the infections in dogs were caused by a relatively genetically stable virus which occurs in eastern

The suitability of selected methods of nucleic acid extraction for detecting Rhodococcus equi DNA in tracheobronchial wash fluid using PCR.

The aim of the present study was to compare the effectiveness of selected DNA extraction methods for the detection of Rhodococcus equi from tracheobronchial wash fluid by PCR. Three methods of nucleic acid extraction were evaluated, based mainly on the activity of proteolytic enzymes. A commercial kit for isolation and purification of bacterial DNA was also used in the study. In one procedure, an additional component, the cationic detergent CTAB, was used. It has been found that the traditional enzyme digestion methods used with the tracheobronchial wash fluid are more suitable to prepare DNA matrix for PCR comparing with commercial DNA isolation kit. Minimum numbers of bacteria detected with the use of traditional enzyme digestion methods and commercial kit were 100 and 500 cells, respectively. Based on the results of the study we can recommend the enzymatic digestion method along with CTAB as an additional component.

Survey of porcine rotavirus G and P genotype in Poland and the United States using RT-PCR.

Porcine rotaviruses are a common cause of gastroenteritis. Several serotypes have been detected based on the two surface proteins VP4 (P-types) and VP7 (G-types). However, limited studies have been performed on the relative frequency of rotavirus types in diarrhetic pigs primarily because of the lack of availability of suitable methods. In this study, we describe a reverse transcriptase-polymerase chain reaction (RT-PCR) method for the typing of P and G types of rotavirus. This method allowed to detect G and P types in 96.8 and 87.1% of isolates collected in the United States, respectively and in 54.5 and 38.6% of isolates collected in Poland, respectively. Within the US specimens the G3, G4, G5, G9 and G10 types were detected in combination with P6 and P7 types while among Polish specimens only G3, G4 and G5 types in combination with P6 and P7 types were identified. In both instances the G4 and G5 were the most prevalent types. These studies show that a RT-PCR typing method is suitable for molecular epidemiological studies and that there is more diversity among porcine rotavirus than previously reported.

Bacterial flora in foals with upper respiratory tract infections in Poland.

Bacteria isolated from nasal cavity of 80 foals with upper respiratory tract infection, as well as from 20 healthy foals, were examined. Within the group of sick animals, from 18 (22.5%) bacteria with recognized pathogenicity were isolated. Coagulase-negative staphylococci and Acinetobacter sp. were the dominant species identified (100 and 45%, respectively). No bacteria species with recognized pathogenicity were isolated from the group of healthy animals. Three cases of death within the group of sick foals were investigated. Rhodococcus equi in two cases and Klebsiella pneumoniae pneumoniae together with Escherichia coli were isolated post-mortem from lung abscesses.

Comparison of polymerase chain reaction and dot hybridization with enzyme-linked immunoassay, virological examination and polyacrylamide gel electrophoresis for the detection of porcine rotavirus in faecal specimens.

The purpose of this study was the evaluation of RT-PCR (reverse transcription-polymerase chain reaction) technique and dot blot hybridization with PCR-generated probe for the detection of group A rotavirus in faecal samples derived from diarrhoeal piglets. They were compared with virological examination (isolation of the virus), polyacrylamide gel electrophoresis (PAGE) and ELISA. The specificity and sensitivity of each assay was assessed against a 'gold standard' which was created on the basis of the virological examination and PAGE results. One hundred and seventeen faecal samples taken from piglets with the signs of diarrhoea were used as research material. cDNA probe labelled with digoxigenin, complementary to the region between 1 and 650 nucleotides of the segment 9 of porcine OSU and Gottfried reference strains was used. The probe detected in the dot blot hybridization 1 ng of dsRNA of the reference porcine strains. Using that test it was shown that 23 positive samples out of 117 samples of diarrhoeic piglets were detected. The RT-PCR technique appeared to be the most specific and sensitive diagnostic method. From 117 amplified faecal samples, the products of RT-PCR reactions were obtained from 24 samples. By means of this method it was possible to find 104 virions per 1 g of sample. Overall comparison of the results showed respective sensitivities and specificities of 100% and 98.9% for RT-PCR, 95.7 and 98.3% for hybridization, 100 and 94.7% for ELISA, 78.3 and 100% for virological examination, 91.3 and 100% for PAGE.

Evaluation of different methods (ELISA, IF, EM, PAGE) for the diagnosis of rotavirus infection in piglets.

The study on the prevalence of rotaviral infections among diarrhoetic piglets in 11 commercial farms in Poland was carried out on 531 faecal samples tested using an ELISA to screen for rotavirus. Of these, 169 were found to contain rotavirus antigen (32% of all cases of diarrhoea tested). Those pig herds managed using the Bisprol system had a lower prevalence of rotavirus in pigs with diarrhoea than those faecal samples obtained from Gi-Gi or Agrokomplex Agard farms. To further establish the presence of rotavirus antigen, 28 of those positive in ELISA samples were taken for isolation of the virus using tissue culture; 18 (64%) isolates were successfully adapted into MA-104 cells and the presence of rotavirus confirmed by immunofluorescence (IF) and electron microscopy (EM). In addition, an analysis of the band patterns visualised on PAGE showed 9 distinct electropherotypes for rotaviral dsRNA among the tested specimens. These findings suggest that rotavirus may represent an important contribution to the incidence of diarrhoea in Polish pig herds. The use of ELISA technology provided an efficient and effective means of evaluating the presence of rotavirus antigen in faecal samples and indicates that this procedure is a very useful tool in epidemiological studies, but that other techniques are required to confirm the presence of virus.