Polish Journal of Veterinary Sciences

Seminal plasma (SP) proteins are responsible for sperm functional quality. Developing a reliable method to determine the degree of oxidative damage of these proteins is important for establishing semen fertilizing ability. The main aim of the study was to verify the applicability of protein carbonyl derivatives measurement in the SP of canine and stallion, using a method with 2,4-dinitrophenylhydrazine (DNPH). The research material consisted of ejaculates obtained from eight English Springer Spaniels, and from seven half-blood stallions during the breeding and non-breeding season. The content of carbonyl groups in the SP was measured on the basis of the reactions with DNPH. The following reagent variants were used to dissolve protein precipitates: Variant 1 (V1) – 6M Guanidine solution and Variant 2 (V2) – 0.1M NaOH solution. It has been shown that to obtain reliable results for the measurement of protein carbonylated groups in the dog and horse SP, both 6M Guanidine and 0.1M NaOH may be used. A correlation was also found between the number of carbonyl groups and the total protein content in the canine (V1: r = -0.724; V2: r = -0.847) and stallion (V1: r = -0.336; V2: r = -0.334) SP. Additionally, the study showed a higher content (p≤0.05) of protein carbonyl groups in the stallion SP in the non-breeding season compared to the breeding season. The method based on the reaction with DNPH, due to its simplicity and cost effectiveness, appears to be suitable for large-scale application in the determination of the SP proteins oxidative damage in dog and horse semen.

A sound knowledge of horseshoe impact on blood flow parameters is required for making shoeing decisions and selecting the most appropriate types of shoes. The aim of this study was to determine the effect of horse shoeing with egg bar shoes and shoes with wedge pads on blood flow parameters in the lateral palmar digital artery measured by Doppler ultrasound. The study was conducted on 16 horses divided into two groups. Horses from group 1 were shod with egg bar shoes. Horses from group 2 were shod with shoes with wedge pads. Doppler ultrasound parameters of the lateral palmar digital artery at the level of the metacarpophalangeal joint were evaluated. Doppler tests were performed before and after shoeing within a monthly interval. The results of the study indicate that egg bar shoes have a greater impact on blood circulation in the distal part of the equine limb than shoes with wedge pads. However, the only parameters to have changed substantially after shoeing with egg bar shoes were end-diastolic velocity (EDV) and mean velocity (Vmn) in the lateral palmar digital artery. A low-resistance blood flow pattern was noted before shoeing. After shoeing in group 1, it remained unchanged in 5 horses, whereas a high-resistance pattern was observed in 3 animals. A low-resistance blood flow pattern was noted in all group 2 horses after shoeing. The difference between the analyzed shoeing techniques could be attributed to increased pressure in the heel bulb area in horses shod with egg bar shoes. Wedge pads shift the load away from the heel bulbs, which might reduce the pressure on the palmar digital vessels and exert a smaller influence on the parameters measured in the Doppler ultrasound test.

Blastocystis spp. is a parasite that causes intestinal infection in humans and other animals. A few studies have been performed in Turkey on the distribution of Blastocystis in cattle. In this study, fecal samples were collected from 100 calves and subjected to analysis based on an SSU rRNA gene fragment. The overall prevalence of the disease was determined as 15% (15/100). This rate was 14.04% for females and 16.28% for males. In addition, three Blastocystis subtypes were identified: ST10, ST14, and novel subtypes ST25. To our knowledge, the ST25 subtype was reported with this study for the first time in Turkey. The nucleotide sequences (OM920832-OM920839) obtained in this study were deposited in GenBank. The results obtained will be useful for a better understanding of the epidemiology of Blastocystis spp., and its effects on public health.

Many viruses are involved in concomitant infections, which are prevalent in nature. In mixed infections, one or both infectious agents may be increased, reduced, or both may be increased while the other is suppressed. Canine distemper virus (CDV) and Canine parvovirus- 2 (CPV-2) are important causes of gastroenteritis in dogs. Detection of these viruses is challenging since the symptoms are very similar. CDV is a member of the morbillivirus genus in the Paramyxoviridae family, and CPV-2 is a member of the Protoparvovirus genus in the Parvoviridae family; and both predominantly affect puppies and induce gastrointestinal symptoms in dogs. The purpose of this study was to contribute to the differential diagnosis of dogs with gastrointestinal symptoms. A PCR technique with specific primers was used to identify CDV and CPV-2 infections in gastroenteric dogs, and clinical changes in the infected dogs were monitored. The VP2 structural gene of CPV and the nucleocapsid gene of CDV were partially amplified in the study. PCR amplified the partial fragments of the CDV nucleocapsid (287 bp) and CPV-2 VP2 proteins (583 bp) from feces. In total, 3 out of 36 stool samples were positive for CDV and CPV-2 in the same dogs. Gasterointestinal symptoms also supported the diagnosis of concomitant infection with CDV and CPV-2 in these dogs. Dehydration and diarrhea in dogs can be signs of various diseases, such as viral, bacterial, and parasitic infections. After the elimination of non-viral pathogens, CDV and CPV-2 should also be simultaneously investigated to establish what is causing these symptoms. This study demonstrates the potential utility of correct diagnosis for the control of viral infection in dogs, but more research with a broader use of PCR-based detections is needed to assess its impact on differential diagnosis for concomitant infections.

Cultivation-based assays represent the gold standard for the assessment of virus infectivity; however, they are time-consuming and not suitable for every virus type. Pre-treatment with platinum (Pt) compounds followed by real-time PCR has been shown to discriminate between infectious and non-infectious RNA viruses. This study examined the effect of Pt and palladium (Pd) compounds on enveloped DNA viruses, paying attention to two significant pathogens of livestock – bovine herpesvirus-1 (BoHV-1) and African swine fever virus (ASFV). Native or heat-treated BoHV-1 suspension was incubated with the spectrum of Pt/Pd compounds. Bis(benzonitrile)palladium(II) dichloride (BB-PdCl ₂) and dichloro(1,5-cyclooctadiene) palladium(II) (PdCl ₂-COD) produced the highest differences found between native and heat- -treated viruses. Optimized pre-treatment conditions (1 mM of Pd compound, 15 min, 4°C) were applied on both virus genera and the heat inactivation profiles were assessed. A significant decrease in the detected quantity of BoHV-1 DNA and ASFV DNA after heat-treatment (60°C and 95°C) and consequent incubation with Pd compounds was observed. BB-PdCl ₂ and PdCl ₂-COD could help to distinguish between infectious and non-infectious enveloped DNA viruses such as BoHV-1 or ASFV.

The aim of this study was to evaluate the fertility response of dairy cows with anovulation type I on repeated low doses of GnRH agonist buserelin. The study was conducted on 83 anovulatory and 60 cyclic Polish Holstein Friesian cows. Anovulation type I was defined as small ovaries with follicles of ≤ 5 mm in diameter and without corpus luteum on two examinations in a 7-10 day interval between 50–60 days after parturition. Cows from the experimental group (n=58) received 0.4 μg of buserelin i.m. once a day for 5 consecutive days. Cows from the negative control group (n = 25) received saline. Sixty cyclic cows receiving no treatment served as positive controls. Intervals from calving to estrus and from calving to conception, pregnancy rate 30-35 days and 260 days after AI, and pregnancy loss were calculated. The anovulatory cows had a substantially prolonged calving to conception interval, decreased pregnancy rate and increased pregnancy loss and culling rate compared to cyclic herd mates. The average calving to conception interval was significantly (p<0.05) shorter in treated cows compared to non-treated anovulatory cows (153.7 days vs 209.3 days). In conclusion, repeated low doses of GnRH analogue buserelin led to a significant shortening of calving to conception interval. More clinical trials are needed to determine the practical usefulness of this method for the treatment of anovulation type I in dairy cows.

Studies on serum interleukin (IL)-31 levels in dogs with atopic dermatitis (AD) and their correlation with disease severity are limited. To the author’s knowledge, there are no studies that measured serum IL-31 in dogs treated with lokivetmab injections, a selective inhibitor of this key cytokine in pruritus. The aim of the study was to evaluate serum IL-31 levels in dogs treated with lokivetmab and correlate it with the severity of canine atopic dermatitis using the pruritus visual analog scale (pVAS) and canine atopic dermatitis extent and severity index (CADESI-04). Ten client-owned dogs diagnosed with AD received two injections of lokivetmab four weeks apart. Disease severity was assessed using the pVAS and CADESI-04 scores before and after both injections. In addition, canine serum IL-31 levels were measured at the same moments. Serum IL-31 was detected in all dogs in the study. There was a significant reduction in pVAS scores and serum IL-31 after administrations. However, there was no difference in CADESI-04 scores, and there was no significant correlation between CADESI-04 scores and serum IL-31 in dogs diagnosed with AD. Nonetheless, a significant positive correlation was observed between the pVAS scores and serum IL-31 levels with lokivetmab therapy, which reinforces the role of IL-31 in the pathogenesis of pruritus in dogs with AD. The data presented here provide further evidence that IL-31 is directly involved in pruritus pathogenesis in dogs with AD. In addition, blocking IL-31 has a significant antipruritic effect, but has no influence on skin lesion severity and extension.

Salmonella contamination in poultry feed is one of the main issues in poultry industry and public health. The aim of the present study was molecular detection and typing of Salmonella serotypes isolated from poultry feeds. Moreover, we determined the antibiotic resistance pattern and the ability of biofilm formation in the serotypes. To this end, eighty feed samples were collected from aviculture depots. Salmonella serotypes were identified by culture and PCR methods. For serological identification, a slide agglutination test was used. BOXAIR and rep-PCR methods were applied to evaluate the diversity of serotypes. The disc diffusion method was performed to evaluate the antibiotic susceptibility of serotypes to sixteen antibiotics. Biofilm formation was also assessed by the microtiter-plate test. From a total of 80 feed samples, 30 samples were contaminated with Salmonella spp., which were divided into 5 different serotypes belonging to B, C, and D serogroups. BOXAIR-PCR (D value [DI] 0.985) and rep-PCR (DI 0.991) fingerprinting of isolates revealed 23 and 19 reproducible fingerprint patterns, respectively. A higher antibiotic resistance was observed to ampicillin and doxycycline (100% each), followed by chloramphenicol (83.33%) and tetracycline (73.33%). Multidrug resistance (MDR) was detected in all Salmonella serotypes. Half of the serotypes possessed the ability of biofilm formation with varied adhesion strengths. These results revealed the high and unexpected prevalence of Salmonella serotypes in poultry feed with MDR and biofilm formation ability. BOXAIR and rep-PCR revealed a high diversity of Salmonella serotypes in feeds and subsequently indicated variation in the source of Salmonella spp. The unknown sources harboring high diversity of Salmonella serotypes indicated poor control, which could cause problems for feed manufacturing.

Testicular torsion is a frequently encountered clinical condition that requires urgent treatment. The aim of this study is to investigate the efficacy of Anise (Pimpinella anisum L.) in treating the pathological condition due to ischemia and reperfusion injury by using biochemical, histopathological and immunohistochemical methods.
A total of 6 groups were formed with 8 male Wistar Albino rats in each group. Group 1 (n=8): control group, Group 2 (n=8): Anise aqueous solution was given orally 5 ml/kg by gavage for 30 days. Group 3 (n=8): Ischemia and Reperfusion (I/R) group, bilateral testicles were rotated 270° and reperfused after 30 minutes of ischemia. Group 4 (n=8): I/R+ Anise group, Group 5 (n=8): Anise+ I/R group and Group 6 (n=8): Anise+ I/R+ Anise group.
The results of the Anise group and the Control group were similar. However, the damage in the I/R group was considerably more severe than in any of the other study groups. While it was observed that spermatogenic cells started to regenerate in the I/R+Anise group, edema and congestion were observed in the Anise+I/R group. In the Anise+I/R+Anise group, all histological findings and biochemical parameters were similar to those of the control group.
It was observed that anise had protective effects in ischemia and reperfusion injury in rat testicles.

Yeast infections such as otitis externa and seborrheic dermatitis in dogs and cats are frequently associated with Malassezia pachydermatis secondary infection. It is part of the normal cutaneous microflora of most warm-blooded vertebrates, however, under certain conditions, it can become a causative agent of infection that needs to be treated pharmacologically. Azole derivatives are the drugs of the first choice. An interesting trend in developing resistance is the use of natural substances, which include manuka honey with confirmed antimicrobial properties. The main intention of this research was to evaluate the mutual effect of manuka honey in combination with four conventional azole antifungals – clotrimazole, fluconazole, itraconazole, and miconazole – on 14 Malassezia pachydermatis isolates obtained from dogs and 1 reference strain. A slightly modified M27–A3 method (CLSI 2008) and the checkerboard test (Nikolić et al. 2017) were used for this purpose. Our results show an additive effect of all 4 antifungals with manuka honey concurrent use. Based on the determined values of fractional inhibitory concentration index (FICI – 0.74±0.03 when manuka honey combined with clotrimazole, 0.96±0.08 with fluconazole, 1.0±0 with miconazole and 1.16±0.26 with itraconazole), it was found in all cases that the effect of substances used is more pronounced in mutual combination than when used separately.

Lipopolysaccharide (LPS), a core part of gram-negative bacteria, is crucial for inducing an inflammatory response in living things. In the current study, we used LPS from Salmonella to stimulate chicken macrophages (HD11). Proteomics was used to investigate immune-related proteins and their roles further. Proteomics investigation revealed 31 differential expression proteins (DEPs) after 4 hours of LPS infection. 24 DEPs expressions were up-regulated, while seven were down-regulated. In this investigation, ten DEPs were mainly enriched in S. aureus infection, complement, and coagulation cascades, which were all implicated in the inflammatory response and clearance of foreign pathogens. Notably, complement C3 was shown to be up-regulated in all immune-related pathways, indicating that it is a potential protein in this study. This work contributes to a better understanding and clarification of the processes of Salmonella infection in chickens. It might bring up new possibilities for treating and breeding Salmonella-infected chickens.

Highly immunogenic nucleotide fragments from 3 genes of Mycoplasma hyopneumoniae strain 232 were selected using information software technology. After repeating each fragment three times, a total of 9 nucleotide fragments were joined together to form a new nucleotide sequence called Mhp232_1092bp. Mhp232_1092bp was directly synthesized and cloned into a pET100 vector and expressed in Escherichia coli. After purification, the proteins were successfully validated by SDS-PAGE and Western blotting using mouse His-tag antibody and pig anti-Mhp serum. BALB/c mice were intraperitoneally injected with purified proteins in the high-dose (100 μg), medium-dose group (50 μg) and low-dose (10 μg) groups. Mice in each group were injected on day 1, day 8 and day 15 of feeding, respectively. Serum samples were collected from all mice on the day before immunization and on day 22 after immunization. The antibody level in the mouse serum was detected using western blotting using purified expressed proteins as antigens. IL-2, TNF-α and IFN-γ were simultaneously detected in the mouse serum by ELISA. The results showed that the 60 kDa protein was successfully expressed and reacted specifically with the specific serum Mhp His-Tag mouse monoclonal antibody and pig anti-Mhp serum. From day 0 to day 22 of immunization, IFN-γ increased from 269.52 to 467.74 pg/mL, IL-2 increased from 14.03 to 145.16 pg/mL, and TNF-α increased from 6.86 to 12.37 pg/mL. The IgG antibody in mice increased significantly from 0 day to day 22 after immunization. This study suggests that the expressed recombinant protein may serve as one of the novel vaccine candidates for Mhp.

Antibiotics are used for postsurgical wound healing purposes but unfortunately, resistance against them demands some alternatives for quick recovery. Sepsis of wounds is a challenge for medical as well as veterinary professionals. Nanoparticles have significant advantages in wound treatment and drug resistance reversal. This study was conducted to appreciate emerging alternates of antibiotics like zinc oxide nanoparticles and plant extracts in topical application. Zinc oxide is considered a good wound healer and its nanoparticles are easy to access. So, the efficacies of zinc oxide nanoparticles and sweet flag plant extract ointments were tested to compare modern and traditional therapeutics as sweet flag is considered a pure medicinal plant. Rabbits were selected for this study due to the healing properties of their skin. Wounds were inflicted on the thoracolumbar region and treated for 29 days post-surgically daily with normal saline and the ointment of zinc oxide nanoparticles and sweet flag extract ointment, prepared in a hydrophilic solvent. Wound shrinkage was observed daily and histopathological analysis was made and results were compared. Zinc oxide nanoparticles ointment showed the most satisfactory results for every parameter included in the study. No side effects of its topical application were observed. Healing was normal without any complications. The preparations of zinc oxide nanoparticles may help in the era of antibiotic resistance as topical drugs in the future.

The aim of the study was to develop a method of laparoscopic embryo transfer in pigs and to compare different variants of this method. Two catheter diameters (1.6 mm and 1.0 mm), the method and site of embryo deposition (oviduct or uterus), the embryo development stage (2 – 4 cell or blastocyst), the method for oviduct or uterus stabilization, the potential for cryopreserved embryo transfer, the developmental potential of the embryos after transfer to the oviduct, patomorphology of the oviduct after transfer and possible clinical complications were taken into consideration. Two studies compared two variants of transfer to the uterus, and five variants of transfer to the fallopian tube. The transfer of embryos by the infundibulum may be of limited use due to handling problems and very low efficiency (pregnancy was not achieved). Very low efficiency was shown after transfer of vitrified embryos. Transfer to the fallopian tube by puncture of the fallopian tube, regardless of the developmental stage of the embryo, is the recommended method of embryo transfer. The histopathological examination of the fallopian tube revealed possible changes within the puncture site. The numerous clinical complications observed did not affect the effectiveness of the method.

A separate, cat-specific hospitalization room away from dogs is recommended to reduce stress in cats; however, this can be difficult for some hospitals to provide. In such cases, measures are undertaken to reduce the cat’s stress by providing a place to hide. However, inability to observe the cat’s condition may be an obstacle to providing veterinary care. The use of a one-way mirror to create a sheltered environment while allowing observation of the cats was assessed. Five healthy cats were assessed using the Cat Stress Score (CSS) while in a cage with either a transparent panel or a one-way mirror. No significant differences in the CSS between the transparent panel and one-way mirror were observed. Variations in the CSS scores depended on the cat’s personality, with friendlier and more sociable cats showing a lower CSS with the one-way mirror. A one-way mirror may be useful to reduce stress in hospitalized cats.

This is the first study to identify 23 protein spots corresponding to 13 proteins in mitochondria isolated from rabbit epididymal spermatozoa. In the group of protein spots identified in stress-induced samples, the abundance of 20 protein spots increased, whereas the abundance of three protein spots (GSTM3, CUNH9orf172, ODF1) decreased relative to the control. The results of this study provide valuable inputs for future research into the molecular mechanisms implicated in pathological processes during oxidative stress (OS).

CVM ( Complex Vertebral Malformations) and Brachyspina (BY) are the most common autosomal recessive genetic defects occurring in the last two decades in Holstein dairy cattle around the world. Beginning from 2004 and 2014, 3035 and 338 Polish Holstein-Friesian bulls were tested to find carriers of CVM and BY, respectively. Among analyzed bulls 191 CVM carriers (6.29%) and 20 BY carriers (5.92%) were identified. No CVM carriers were observed beginning from 2016, whereas only single BY carriers was identified annually for the last 5 years. One bull turned to be double CVM/ BY carrier as a son of also double CVM/BY top Dutch sire (JABOT 90676-4-9). It is shown that CVM and BY defects are practically eradicated from Polish dairy cattle although incidental testing should be continued if new bulls with CVM or BY carriers in sire or dam pedigree will unexpectedly appear.

Zinc oxide (ZnO) has been applied for many years in the production of pigs to reduce the number of diarrhoea in weaned piglets. In June 2022, the European Union banned the use of zinc oxide (ZnO) in pig feed. According to scientific reports, the may reason was the accumulation of this microelement in the environment of pig production. It has been shown that frequent application of ZnO can lead to increased antibiotic resistance in pathogenic swine microflora. The main alternatives to ZnO are probiotics, prebiotics, organic acids, essential oils, and liquid feeding systems.
Alternatives to ZnO can be successfully used in pig production to reduce the number of diarrhoea among piglets during the postweaning period. Additional reports indicated that bacteriophage supplementation has a positive effect on the health of pigs. The article provides an overview of current ZnO substitutes that can be used in pig farming.