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Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Contents The aim of this study was to determine onset of puberty in Cameroon Dwarf goats ( CDGs) kept as pets in northwestern Croatia by determining progesterone ( PGS) and insulin-like growth factor-I ( IGF-I) levels in the blood by RIA methods. The first cyclic ovarian activity was estimated according to hormone profiles as determined in CDG in a moderate climate environment. Sixteen female CDG kids were kept in stables with access to pasture which provided space for exercise. The goat kids born in winter (December-January-February), spring (March-April-May), summer (June-July-August) and autumn (September-October-November) were assigned into four groups according to the season of the year of birth ( n = 4 in each group). At 75 days of age at the initiation of the study, they weighed between 3.2 and 5.1 kg (4.24 ± 0.53 kg). The onset of ovulatory activity was determined by PGS and IGF-I serum concentrations every 10 days starting from 75 days to 155 days of age. The onset of puberty in CDG kids occurred on average at 141.15 ± 2.66 days of age, but varied depending on the season of birth. All CDG kids born during summer were in heat by 155 days of age. In the other groups (kids born in autumn, winter or spring), one goat in each group was not in heat. Changes in blood serum PGS and IGF-I concentrations during prepubertal and pubertal periods could aid in the evaluation of reproductive status and determination of the onset of puberty in CDG during all seasons of the year in a moderate climate region. [ABSTRACT FROM AUTHOR]

The effects of stress induced by adrenocorticotropic hormone (ACTH) on biochemical and immune changes in Swedish Landrace boars aged approximately 6–7 months were observed during ACTH administration and for 16 days after the cessation of treatment. Adrenocorticotropic hormone treated animals ( n = 7) were given intravenously 10 μg/kg body mass of ACTH for 3 days. Control group of animals ( n = 7) received intramuscularly 1 ml of sterile 0.9% saline. Total protein and globulin levels were significantly elevated during the induced stress period and for the next 16 days ( P < 0.01 to P < 0.0001, respectively). Also, serum immunoglobulin IgG concentration was significantly elevated during and after ACTH injection ( P < 0.001 to P < 0.0001). Adrenocorticotropic hormone treatment had no effect on serum albumin, IgA and IgM concentrations. Glucose concentration was significantly decreased on the second day of ACTH administration ( P < 0.001) and on day 9 after treatment ( P < 0.001). Calcium level was significantly decreased only for 24 h after the last ACTH dosage ( P < 0.01). Also, serum phosphorus level was significantly decreased on the first ( P < 0.05) and third ( P < 0.001) days of ACTH challenge but remained unaffected after the cessation of ACTH treatment. It is concluded that the administration of ACTH to boars results in immune humoral and biochemical changes during stress and the post-stress period. [ABSTRACT FROM AUTHOR]

Summary The influence of an attenuated classical swine fever virus C strain vaccine and a subunit E2 vaccine against classical swine fever on the peripheral blood leucocyte proportion and phenotypic expression in 12-week-old pigs was studied. The C strain was amplified in minipig kidney cell culture and final product contained 104±0.15 TCID50 /ml, while the subunit vaccine contained 32 μ g per dose of gp E2. Haematological findings showed that the vaccines did not cause leucopenia or lymphocytopenia and the number of neutrophils and eosinophils during the observation period was within physiological range. The results of the proportion of CD4a+ , CD5a+ , CD8a+ , wCD21+ , CD45RA+ , CD45RC+ , non-T non-B, SWC3a+ and CD11b+ cells were gained by single-colour flow cytometry. At the end of the trial a significantly increase of percentage of CD4+ , CD5a+ , CD8+ , wCD21+ cells has been found in pigs that received the subunit vaccine and the percentage of CD4+ , CD5a+ , CD8+ , CD45RA+ and CD45RC+ cells was higher in pigs that received the attenuated vaccine. Twenty-eight days after vaccination the percentage of CD4+ , CD45RA+ and CD45RC+ was significantly higher in pigs vaccinated with the C strain than in pigs vaccinated with the subunit vaccine. In contrary, the percentage of the wCD21+ cells was higher in pigs that received the subunit vaccine. Statistically higher values of SWC3a+ and lower values of CD11b+ cells was observed in pigs that received the attenuated vaccine than in pigs vaccinated with the subunit vaccine. Taken altogether, our results showed that the subunit vaccine produced a better stimulation of B cells and CD11b+ monocytes/macrophages /granulocytes/NK cells, whereas the attenuated vaccine induced a higher response of Th cells, naive/memory cells and macrophages/neutrophils. Thus, both vaccines were able to influence the porcine immune system, by activating different subsets of the immune effector/accessory cells. [ABSTRACT FROM AUTHOR]

During the hunting season in February 1999, a total of 44 blood samples were collected from wild boars shot in the area of MoslavaCka gora. These blood samples were examined by enzyme immunoassay for the presence of antibodies to classical swine fever (CSFV), Aujeszky's disease (ADV), bovine viral diarrhoea (BVDV), and porcine reproductive and respiratory syndrome (PRRSV) viruses. Out of 44 serum samples examined, 17 (38.63%) were positive for CSFV, 24 (54.54%) were positive for ADV and two (4.54%) were positive for BVDV. All sera were negative for PRRSV. The results, recorded for the first time in Croatia, supported the hypothesis that wild boar act as a potential reservoir of CSFV, ADV and BVDV, and thus have a role in the epidemiology of these diseases. [ABSTRACT FROM AUTHOR]

Abstract Background Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Findings Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Conclusions Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

The effects of radiofrequency electromagnetic radiation (RF-EMR) on semen quality have been in the spotlight in recent years, though research results to date have been contradictory. The effects of RF-EMR amongst others depend upon frequency, and there is currently no literature concerning the influence of 5G frequencies on both DNA integrity and spermatozoa vitality in males. The aim of this study was to investigate the effect of 5G RF-EMR on sperm membrane integrity, mitochondrial potential, and DNA integrity of in vitro exposed semen of breeding boars. The study included semen samples of eight breeding boars of the Pietren breed and four breeding boars of the German Landrace breed, from 1.5 to 3.5 years in age. Freshly diluted semen of each boar was divided into a control (n = 12) and experimental group (n = 12). The samples of the experimental group were exposed for 2 hours to continuous RF-EMR at a single frequency (700 MHz, 2500 MHz and 3500 MHz) and an electromagnetic field strength of 10 V/m using a transverse gigahertz electromagnetic cell. Sperm DNA fragmentation was assessed using a Halomax® kit and sperm membrane integrity and mitochondrial potential was assessed using a PI⁄SYBR-14 LIVE⁄DEAD viability kit with JC-1. A significantly higher proportion of spermatozoa with DNA fragmentation was found in exposed semen samples for all frequencies compared to the control group. The highest DNA damage was recorded in semen samples exposed to 5G RF-EMR at 2500 MHz (p < 0.01) and 3500 MHz (p < 0.05) vs. control semen samples. A significantly higher proportion of spermatozoa with damaged cell membrane and good mitochondrial potential was recorded in semen samples exposed with 3500 MHz. In vitro exposure of breading boar semen to 5G RF-EMR significantly increases the proportion of DNA fragmentation. The harmful effect of 5G RF-EMR on the proportion of spermatozoa with damaged DNA was frequency dependent. The 3500 MHz frequency displayed the most harmful effects due to significant impacts on DNA integrity and spermatozoa vitality indicators., Competing Interests: Declaration of competing interest The authors declare no potential conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Anthropogenic radiofrequency electromagnetic radiation (RF-EMR) from wireless technologies has increased dramatically. The boar semen used for artificial insemination is essential in sustaining the pig industry, and additionally it is also exposed to the effects of the RF-EMR of wireless technologies. Furthermore, there are no data on the effects of RF-EMR on semen quality, and this is the first analysis of sperm's morphometric parameters for assessing the effect of RF-EMR on the spermatozoa subpopulations of boars. This study investigated the effect of RF-EMR on in vitro exposed breeding boar semen spermatozoa motility and the proportions of spermatozoa subpopulations according to their morphometric head and tail parameters. The semen samples of 12 boars were divided into control and experimental groups. The samples in the experimental group were exposed in a gigahertz transverse electromagnetic chamber at a frequency of 2500 MHz (the frequency band used in 5G technology) and an electric field strength of 10 Vm -1 for two hours. After exposure, the spermatozoa motility was evaluated for both groups. A morphometric analysis of the semen smears was performed using SFORM software (Version 1.0; VAMS, Zagreb, Croatia). The progressive spermatozoa motility was significantly reduced in the experimental group (74.7% vs. 85.7%). PC analysis and cluster analysis revealed two spermatozoa subpopulations: S1, spermatozoa with a more regular head shape and a smaller midpiece outline, and S2, spermatozoa with a more elongated head shape and a larger midpiece outline. The experimental semen samples had a greater proportion of the S1 spermatozoa subpopulation (68.2% vs. 64.4%). The effect of RF-EMR at 2500 MHz on the in vitro exposed boar semen resulted in decreased progressive spermatozoa motility and a lower proportion of the spermatozoa subpopulation with a higher fertilizing potential.

The objective of this study was to evaluate the effects of in-feed clinoptilolite (CPL) on serum metabolic and antioxidative biomarkers, acute phase proteins and reproductive performance in cows during pregnancy and lactation. A total of 78 Holstein-Friesian cows were randomly assigned into two groups: the treatment group, cows fed CPL (n = 38) which received 50 g of powdered CPL twice a day from day 180 before parturition to day 60 postpartum; and the control group (n = 40). Blood samples were taken on days 180, 90, 60, 30 and 10 before parturition, on day of calving and on days 5, 12, 19, 26, 33, 40 and 60 postpartum, and were analysed for metabolic biomarkers: glucose, triglycerides, total cholesterol, high density lipoprotein cholesterol, non-esterified fatty acids, beta-hydroxybutyrate (BHB), antioxidative biomarkers and acute phase proteins: paraoxonase-1 (PON1), apolipoprotein A-I, haptoglobin (Hp) and serum amyloid A (SAA). CPL supplementation increased concentration of glucose and significantly decreased (P < .05) level of BHB during puerperium. The SAA concentration in CPL-fed cows was significantly decreased (P < .05) on days 33, 40 and 60 postpartum as well as Hp concentration on days 0 and 12 postpartum. The results of this study suggest that the CPL-fed cows may have improved metabolic status due to the tendency of greater glucose levels and decreased BHB values during early lactation. In addition, acute phase response was lower (P < .05) in CPL-fed cows. Such an outcome might be attributed to the effect of dietary CPL on intensity and severity of the negative energy balance and inflammatory response in dairy cows., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The insoluble immune complexes (ICs) were prepared under the conditions of double immunodiffusion in gel, using the suspension of the ultrasound treated PK-15 cell-line infected with porcine parvovirus (PPV) containing both viral particles and viral proteins, as well as pig or rabbit anti-PPV polyclonal immune sera. The immunodiffusion performed in an agarose gel allows only viral subunits with a molecular mass equal to or less than 1000 kDa, rather than the viral particles, to diffuse through the gel and reach the point where the immunoprecipitate is to be formed. The immunoprecipitation under the conditions of the diffusion ensures the optimal, i.e. equimolar ratio of both immunoprecipitating components, antibody/antigen in the IC. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the Western blot analyses showed the ICs were composed of two proteins, a protein in which molecular mass corresponded to the VP2 of the PPV and a protein with a molecular mass of the IgG. This suggests that the ICs are mainly composed of the VP2 antigen and IgG class antibodies. The potency of the IC-vaccines prepared in the form of a water-in-oil-in-water emulsion was compared with that of a commercially available, inactivated oil vaccine. The vaccination of gilts, 6 weeks before mating, with the IC containing allogeneic pig antibodies, resulted in the development of high and long-lasting anti-PPV antibody titres, similar to those generated by the licenced vaccine (P > 0.01). The content of the virus material administered by the IC was twice lower than that in the licenced vaccine. Neither systemic nor local reactions were observed in the gilts during the period of the trial with the IC vaccine. The number of viable piglets per litter varied between 9 and 12 and no signs of the PPV infection were detected. Rabbits were used as one of the alternative laboratory animal models accepted for the testing of the vaccine against the PPV. The rabbit humoral immune response generated by the IC containing the allogeneic antibodies were higher than that generated by the ICs containing the xenogeneic pig antibodies. It was similar to that generated by two-times higher content of the virus material administered by a commercially available vaccine. The IC-based vaccines belong to non-replicating, subunit vaccines, which are both ecologically convenient and the safest vaccines of all.

The effects of stress induced by adrenocorticotropic hormone (ACTH) on biochemical and immune changes in Swedish Landrace boars aged approximately 6-7 months were observed during ACTH administration and for 16 days after the cessation of treatment. Adrenocorticotropic hormone treated animals (n = 7) were given intravenously 10 microg/kg body mass of ACTH for 3 days. Control group of animals (n = 7) received intramuscularly 1 ml of sterile 0.9% saline. Total protein and globulin levels were significantly elevated during the induced stress period and for the next 16 days (P < 0.01 to P < 0.0001, respectively). Also, serum immunoglobulin IgG concentration was significantly elevated during and after ACTH injection (P < 0.001 to P < 0.0001). Adrenocorticotropic hormone treatment had no effect on serum albumin, IgA and IgM concentrations. Glucose concentration was significantly decreased on the second day of ACTH administration (P < 0.001) and on day 9 after treatment (P < 0.001). Calcium level was significantly decreased only for 24 h after the last ACTH dosage (P < 0.01). Also, serum phosphorus level was significantly decreased on the first (P < 0.05) and third (P < 0.001) days of ACTH challenge but remained unaffected after the cessation of ACTH treatment. It is concluded that the administration of ACTH to boars results in immune humoral and biochemical changes during stress and the post-stress period.

Serologic evidence of exposure to porcine parvovirus (PPV) in the wild boar (Sus scrofa) in Croatia was investigated. Serum samples from 219 wild boars captured during 2003 from 12 different locations in the Republic of Croatia were tested by using a commercial enzyme-linked immunoassay (ELISA) and a hemagglutination inhibition (HI) test. Antibodies to PPV were detected in 91 (41.6%) of tested samples and positive results were detected in wild boar from all sample locations. Adults had a significantly higher prevalence (70%) than juveniles (31%; P < 0.01). Our results indicate that wild boar populations throughout the Republic of Croatia are exposed to PPV.

In December 2001, bluetongue (BT) was confirmed serologically by the Croatian Veterinary Institute using the competitive enzyme-linked immunosorbent assay (c-ELISA). Results of the serological testing of blood samples from ruminants in the Dubrovacko-Neretvanska County are presented (3,318 sera of ruminants from 53 herds were examined). In total, 357 bovine sera (178 or 49.9% positive), 1,268 ovine sera (174 or 13.7% positive) and 1,693 caprine sera (270 or 15.9% positive) were tested. Antibodies to BT virus serotype 9 were detected in 212 of the positive sera by serum neutralisation. A preliminary light-trap survey for midges of the Culicoides genus was also performed in the Dubrovacko-Neretvanska County. Fourteen light-trap collections from seven locations were examined and yielded a total of 4,872 Culicoides of which 4,492 (92%) belonged to the Obsoletus Complex (including C. obsoletus and C. scoticus).

The influence of an attenuated classical swine fever virus C strain vaccine and a subunit E2 vaccine against classical swine fever on the peripheral blood leucocyte proportion and phenotypic expression in 12-week-old pigs was studied. The C strain was amplified in minipig kidney cell culture and final product contained 10(4 +/- 0.15) TCID50/ml, while the subunit vaccine contained 32 microg per dose of gp E2. Haematological findings showed that the vaccines did not cause leucopenia or lymphocytopenia and the number of neutrophils and eosinophils during the observation period was within physiological range. The results of the proportion of CD4a+, CD5a+, CD8a+, wCD21+, CD45RA+, CD45RC+ , non-T non-B, SWC3a+ and CD11b+ cells were gained by single-colour flow cytometry. At the end of the trial a significantly increase of percentage of CD4+, CD5a+, CD8+, wCD21+ cells has been found in pigs that received the subunit vaccine and the percentage of CD4+, CD5a+, CD8+, CD45RA+ and CD45RC+ cells was higher in pigs that received the attenuated vaccine. Twenty-eight days after vaccination the percentage of CD4+, CD45RA+ and CD45RC+ was significantly higher in pigs vaccinated with the C strain than in pigs vaccinated with the subunit vaccine. In contrary, the percentage of the wCD21- cells was higher in pigs that received the subunit vaccine. Statistically higher values of SWC3a+ and lower values of CD11b+ cells was observed in pigs that received the attenuated vaccine than in pigs vaccinated with the subunit vaccine. Taken altogether, our results showed that the subunit vaccine produced a better stimulation of B cells and CD11b+ monocytes/macrophages /granulocytes/NK cells, whereas the attenuated vaccine induced a higher response of Th cells, naive/memory cells and macrophages/neutrophils. Thus, both vaccines were able to influence the porcine immune system, by activating different subsets of the immune effector/accessory cells.

During the hunting season in February 1999, a total of 44 blood samples were collected from wild boars shot in the area of Moslavacka gora. These blood samples were examined by enzyme immunoassay for the presence of antibodies to classical swine fever (CSFV), Aujeszky's disease (ADV), bovine viral diarrhoea (BVDV), and porcine reproductive and respiratory syndrome (PRRSV) viruses. Out of 44 serum samples examined, 17 (38.63%) were positive for CSFV, 24 (54.54%) were positive for ADV and two (4.54%) were positive for BVDV. All sera were negative for PRRSV. The results, recorded for the first time in Croatia, supported the hypothesis that wild boar act as a potential reservoir of CSFV, ADV and BVDV, and thus have a role in the epidemiology of these diseases.

Enzyme linked immunosorbent assays (ELISAs) and a nested polymerase chain reaction after reverse transcription (RT-PCR) were used for the detection of the Chinese strain (C strain) of classical swine fever virus (CSFV) in blood and tissue samples of experimentally inoculated piglets. One group of 10 piglets was inoculated with C strain material from rabbits and a second one with material from infected minipig kidney (MPK) cell culture. Tested blood samples were taken on the day of inoculation as well as on days 2, 4, 6, 8, 10, 13 and 16. Samples of spleen, tonsil and brain tissue were collected from piglets on days 6, 8, 10, 13 and 16 and tested for glycoprotein E(RNS) and protein NS2-3 using commercially available ELISA kits. E(RNS) and NS2-3 were detected earlier in blood samples of piglets inoculated with the C strain propagated in a cell culture. Regardless of propagation the presence of the viral E(RNS) and NS2-3 was detected in spleen and tonsil samples simultaneously. The C strain propagated in a cell culture was found in only one brain sample, whereas, the virus propagated in rabbits was detected in 70% of the brain samples. For the detection of the CSFV RNA in blood samples, a part within the 5' non-coding region was amplified. The differences in the results gained by antigen detection in blood samples decreased when nested RT-PCR was used.

Research was carried out into the effect that different quantities and compositions of concentrated portions of meal had on certain haematological properties and on the immune response of mares in the course of hyper-immune antitetanus sera production. The experiment involved 24 Nonius and Lipizzaner cross-bred mares divided into two groups of 12 animals each, a control group and a trial group. The experiment lasted 12 months, with haematological and immunological tests being carried out every 30 days. During the course of the experiment each mare was subjected to 11 immunisation cycles, and in that period no mare fell ill. Leukocyte and haematocrit counts revealed no differences between the control and the test groups throughout the entire duration of the experiment. Following the second immunisation cycle the leukocyte count increased in both groups of mares, remaining significantly higher than the normal physiological value in horses throughout the experiment. The concentrated portion of meal, differing both in quality and quantity, had no influence on leucocyte count and haematocrit level in the mares throughout the experiment. However, the immunodiffusion titre in the test group did show a slightly higher value of ID-titre than in the control group, indicating that the quantitatively and qualitatively differing portions of meal had a more positive effect on immune response and anti-tetanus anti-body titre in the test group than it did in the control group.