Ocular abnormalities in Whippets practicing lure coursing in Poland.

Coursing is a canine sport discipline invented for sighthounds. The dogs chase the mechanically operated lure on an open area at the track that consists of straight lines as well as turns. Thus, the dogs must rely mainly on their sight in order to chase the lure and to compete. Whippets are the most popular sighthound breed in Poland and more and more dogs practice lure coursing on professional level. Several ocular disorders are known or presumed to be inherited in Whippets, so we decided to check how often they occur amongst competing dogs and if they have any impact on the results during competitions. Forty-five regularly competing Whippets underwent complete ophthalmic examination, including evaluation of menace response, pupillary light reflex and dazzle reflex, biomicroscopic examination conducted before and after dilating pupils with topical tropicamide and fundus examination with indirect ophthalmoscopy. Refractive state of each eye was assessed via streak retinoscopy. Ocular abnormalities were identified in 24 dogs (53.3%). The most common finding was vitreous degeneration identified in 9 dogs (20%), five dogs presented multiple lesions. There were no statistically significant differences regarding sex, age, and ranking points between the dogs with and without lesions. However, some of the identified lesions are likely to progress, so we postulate that the ophthalmological examination should receive more attention in sport Whippets, to check if it has any impact on competing dogs at more advanced age.

The lack of the influence of various species of Mycoplasma spp. on canine semen quality.

Mycoplasmas colonize fish, reptiles, birds and mammals, being commensals or causing diseases, sometimes severe in ruminants, swine, poultry, or wildlife animals. So far, 15 species of canine Mycoplasma spp. have been described. Conflicting results have been presented regarding the pathogenicity of Mycoplasma spp. Although many virulence factors of these bacteria have been described, they still require attention. The main aim of our study was to evaluate the presence of known canine Mycoplasmas in the male reproductive tract of clinically healthy dogs. The second aim was to check if Mycoplasma spp. cause any abnormalities in semen quality that could have further consequences and to propose the schemes for managing the carriers. 83.3% of examined dogs were Mycoplasma spp. -positive dogs, and most of them were the carriers of more than one species. Six dogs had azoospermic ejaculates. The total spermatozoa numbers were similar in Mycoplasma -positive and negative groups. Motility was slightly higher in Mycoplasma spp.-negative group, but the difference was not statistically significant. There was no significant difference in semen characteristics between the carriers and Mycoplasma spp.-negative dogs. Neither the individual species nor the number of species strains had a significant effect on sperm morphological parameters as well as viability. Semen quality parameters are not correlated with the species found on the prepuce. Over 70% Mycoplasma spp.- positive dogs have more than one species of this bacteria. Despite finding mycoplasmas in azoospermic dogs, we suggest that they were not the cause of infertility. Mycoplasma spp. could be a part of normal microbiota in canine prepuce in individuals without any clinical signs.

Stress response after race and endurance training sessions and competitions in Arabian horses.

Assuring a high level of animal welfare is a critical aspect of contemporary animal husbandry. Equine athletes begin their careers at a very young age when they are still developing and they are both physically and mentally immature. Lack of scientific knowledge of the stress related to horse racing impedes the development of optimal training programs to attain equilibrium between the best sport results and optimal welfare. This study aimed to determine the influence of the intensity and type of physical activity on peripheral blood cortisol concentration. Thirty untrained Arabians, 9 endurance and 21 race horses were enrolled in this longitudinal study. Blood samples were analysed every 3-weeks in 4 training sessions and 2 races in racing horses and monthly after 2 training sessions and 1 competition in endurance horses. Cortisol concentration was measured at rest and 30 min. after physical effort. Racing horses were divided into two groups of the best and the worst performers. Cortisol concentration increased significantly after training and competition, however both in racing and endurance horses the increase was more intensive after competition. In the racing horses, cortisol concentration tended to gradually increase after the subsequent trainings during the racing season, however the starting gate did not appear to exert any impact on cortisol concentration. The best performing race horses appeared to have a lesser increase in cortisol concentration after the race than the worst performing horses, however the cortisol concentration after the race was not significantly different between these two groups of horses. This study suggests that an optimal training program can induce a stress response which is likely not to have any harmful impact on an athlete's welfare. The more intense effort associated with competition events, both in race and endurance horses, results in a greater stress response, indicating that these kinds of events should be limited to assure animal welfare. Finally, better performance horses adapt more readily to physical activity as they experience less increase of serum cortisol concentration after the race.

Serum amyloid A in equine health and disease.

Serum amyloid A (SAA) is the major acute phase protein in horses. It is produced during the acute phase response (APR), a nonspecific systemic reaction to any type of tissue injury. In the blood of healthy horses, SAA concentration is very low, but it increases dramatically with inflammation. Due to the short half-life of SAA, changes in its concentration in blood closely reflect the onset of inflammation and, therefore, measurement of SAA useful in the diagnosis and monitoring of disease and response to treatment. Increases in SAA concentration have been described in equine digestive, reproductive and respiratory diseases and following surgical procedures. Moreover, SAA has proven useful for detection of some subclinical pathologies that can disturb training and competing in equine athletes. Increasing availability of diagnostic tests for both laboratory and field use adds to SAA's applicability as a reliable indicator of horses' health status. This review article presents the current information on changes in SAA concentrations in the blood of healthy and diseased horses, focussing on clinical application of this biomarker.

Whole body and partial body cryotherapies - lessons from human practice and possible application for horses.

Whole body and partial body cryotherapies (WBC and PBC) have been successfully used in human medicine, and currently also are being proposed in veterinary practice.In horses, only the partial body cryotherapy provided in cryosauna is considered, due to the technical conditions. These therapies have been dedicated to human patients with rheumatic and inflammatory diseases as well as an assistance during training in athletes. The anti-inflammatory effects have been demonstrated clinically and indicated by the changes in several hematological and immunological parameters, however, various patterns have been described, depending on the protocols and the treating subjects. The numbers of white blood cells and the differential counts either increased or remained unchanged but the cytokine concentrations generally changed towards various anti-inflammatory profiles and the modifications of immunological molecules having paracrine effects have been reported.In equine practice, local cold therapies have been successfully used, so the therapy in cryosauna, which allows for a much shorter procedure with much lower temperature seems promising.

The cyclooxygenase-2/prostaglandin E2 pathway and its role in the pathogenesis of human and dog hematological malignancies.

The overexpression of cyclooxygenase-2 (COX-2) has been documented in many types of cancer occurring in humans and animals. Increasing evidences have shown that the overexpression of COX-2 and increased production of prostaglandin E2 (PGE2) correlate with poor prognosis in human solid tumours and hematological malignancies. Both, in vitro and in vivo studies have demonstrated that increased proliferation of cancer cells as well as an impairment of anti-tumour immunity are influenced by the overexpression of this enzyme. In leukemia and lymphoma, an increased activity of COX-2 and subsequent increase in prostaglandins (PGs) concentration allow cancer cells to evade immune response and contribute to metastases. Cancer stem cells (CSCs) in tumour microenvironment, suppression of innate and adaptive immunity depends on COX-2/PGE22 axis activity which increases in hematological malignancies. Cyclooxygenases inhibitors block the formation of PGs, consequently inhibiting angiogenesis, and in some malignancies they decrease cancer cells proliferation and tumour invasiveness. They also increase apoptosis of CSCs and cancer cells, decrease their drug resistance as well as enhance the host immune response. Therefore COX-2/PGE2 axis suppressors: selective COX-2 inhibitors or PG receptors antagonists have been considered as promising anticancer drugs. In comparative oncology dogs are increasingly used as a large animal model because they share the same environmental conditions with people and are exposed to the same environmental factors and also due to their relatively short life span. In dogs, spontaneously occurring non-Hodgkin lymphomas and leukemias have a large number of genetic and morphological features that are similar to those of humans' corresponding cancers. This, additionally makes the species a useful model for the study of new therapeutic strategies in human oncology. While the influence of COX-2 activity and PGE2 receptors have been evaluated extensively in human cancer, their role in veterinary oncology still needs to be elucidated.

Reference intervals for selected hematological and biochemical variables in Hucul horses.

BACKGROUND: Hucul horses are the unique, genetically distinct breed of Carpathian Mountains. Even though they are recognized as primitive breed, many morphological differences between them and other primitive horses have been reported. Neither hematological nor blood biochemical studies in this breed have been conducted so far. OBJECTIVES: The aim of this study was to establish the reference intervals for basic hematological and selected biochemical variables and to compare them with other breeds. MATERIAL AND METHODS: Blood samples were collected from 168 Hucul horses and the analyses were performed using routine methods. Mainly nonparametric method was used to establish reference intervals. RESULTS: The following reference intervals have been established (rounded to two significant digits): RBC: 7.0-13×1012/l; HGB: 106.1-195.8 g/l; HCT: 0.3-0.6 l/l; MCV: 35-50 fl; MCH 11.9-17.1 pg; MCHC: 31.9-34.8 g/dl; WBC: 7.5-22×109/l, bands: 0-0.5×109/l; segmented neutrophils: 3.3-10×109/l; eosinophils: 0-1.1×109/l; basophils: 0-0.3×109/l; lymphocytes: 1.9-12×109/l; monocytes: 0-0.2×109/l; PLT 95-350×109/l; MPV 5.2-7.0; ALP: 98-425 U/l; AST: 220-470 U/l; GGT: 9.1-31 U/l; total bilirubin: 6.5-29 µmol/l; CPK: 120-640 U/l; triglycerides: 0.1-0.9 mmol/l; urea: 3.8-11 mmol/l; creatinine: 44 -140 µmol/l; serum amyloid A: 130-5200 µg/l. CONCLUSIONS: Hematological and biochemical variables in Hucul horses were closer to hot-blooded then to cold-blooded and primitive horses or wild equidae. The reference intervals presented in this study pose clinically useful tool for evaluation of blood check-up in Hucul horses.

Survivin expression in canine lymphomas in relation with proliferative markers.

Survivin is a member of apoptosis inhibiting proteins family. Apart from its antiapoptotic activity it plays a critical role in regulating the cell cycle and mitosis. It is overexpressed in most human malignancies. While the prognostic significance of survivin expression is widely investigated in human non-Hodgkin's lymphomas, little is known about its expression in canine lymphomas. The aim of the study was to evaluate the expression of survivin in canine lymphomas in relation to proliferation markers (mitotic index and percentage of Ki67-positive cells). Survivin was found in all examined lymphomas belonging to 6 different morphological subtypes with nuclear immunoreactivity. In most of lymphomas (18/25) survivin expression ranged 10%-25% of positive cells. Only single cases had lower (0-10% positive cells, 1/25) or higher (25-50% and > 50% positive cells, 5/25 and 1/25, respectively) index of survivin. Neither mitotic index nor proliferative index correlated with survivin expression when the values quantified randomly in whole specimens were compared. However, when survivin expression were quantified in selected tumor areas of low and high proliferation activity the high correlations between survivin expression and proliferation index were found. The results indicated that survivin is commonly expressed in canine lymphomas. Nuclear labelling together with the relation of its expression and proliferative activity in highly proliferative areas of neoplastic tissue suggest a potential role of survivin in cell cycle activation in canine lymphoma cells. However, further studies of the relation between expression of survivin and other proteins involved in cell cycle regulation are needed. Moreover, the results suggest that survivin may pose the therapeutic target in canine lymphomas.

Post-exercise dynamics of serum amyloid A blood concentration in thoroughbred horses classified as injured and non-injured after the race.

The aim of this study was to evaluate serum amyloid A (SAA) concentration in horses with orthopedic injuries acquired during racing and in healthy ones after completing the race. Injuries of bone and tendon did not cause radical increase in SAA concentration observed in other inflammatory conditions. SAA concentration correlated positively with white blood cell count (WBC) on the 3rd-4th days after race being significantly higher in the injured horses than in the control group in that time. It was suggested that racing effort may cause increase in SAA level, more pronounced in horses manifesting clinical signs of orthopedic injury after the race.

Effectiveness of training programmes used in two stables of thoroughbred race horses.

The purpose of this study was to compare the training methods used in two stables and their effects on selected blood parameters and race results. A total number of 36 thoroughbred race horses was examined in two groups, trained by two trainers. Twenty-four horses (group A) were trained at Sluzewiec and the remaining twelve horses (group B) were kept and trained in a private stable. The experiment lasted for five months. The activities of CPK (creatine phosphokinase) and AST (aspartate aminotransferase) and the concentration of LA (lactic acid) were determined. The speed was controlled and recorded by a Garmin GPS system. The analysis of the General Handicap rating demonstrated that the training methods used in stable A were more effective and resulted in better classification of these horses. Training methods in both stables were evaluated and compared on the basis of maximal speeds during training sessions and related post exercise LA concentrations. The main differences between training methods used in both stables concerned the workload and the time of work with the rider. Analysis of the values measured in individual horses from stable B have shown that AST and CK activities were high not only in all young, 2-year-old horses but also in three older ones. This seems to confirm the lack of balance and proper movement coordination in these horses, resulting in high activities of muscle enzymes.

DNA barcoding distinguishes pest species of the black fly genus Cnephia (Diptera: Simuliidae).

Accurate species identification is essential for cost-effective pest control strategies. We tested the utility of COI barcodes for identifying members of the black fly genus Cnephia Enderlein (Diptera: Simuliidae). Our efforts focus on four Nearctic Cnephia species-Cnephia dacotensis (Dyar & Shannon), Cnephia eremities Shewell, Cnephia ornithophilia (Davies, Peterson & Wood), and Cnephia pecuarum (Riley)--the latter two being current or potential targets of biological control programs. We also analyzed one Palearctic species, Cnephia pallipes (Fries). Although Cnephia adults can be identified anatomically to species, control programs target the larval stage, which is difficult or impossible to distinguish morphologically. By using neighbor-joining, maximum parsimony, and Bayesian methods, we found that COI barcodes successfully identified three Nearctic Cnephia species, but not C. pecuarum. The Palearctic C. pallipes was also successfully identified. Despite nonmonophyly of C. pecuarum, we show that data from COI barcoding, in combination with geographical and ecological information, can be used to distinguish all four Nearctic species. Finally, we discussed 1) possible reasons for paraphyly in C. pecuarum, 2) topological concordance to previously reported chromosomal dendrograms, and 3) evolution of diverse feeding strategies within the genus Cnephia.

Serum amyloid A level as a potential indicator of the status of endurance horses.

REASONS FOR PERFORMING STUDY: Changes in serum levels of acute phase proteins (APPs) reflect the acute phase reaction, a rapid and nonspecific response to any tissue damage. Serum amyloid A (SAA) is the main APP in horses, which increases in various diseases, surgical injuries and after long distance endurance rides; however, this nonspecific parameter has not been investigated as an indicator of subclinical disorders, which may result in elimination during endurance competitions. OBJECTIVES: To evaluate the serum concentration of SAA as a potential indicator for the status of horses prepared for long distance endurance rides (120 and 160 km). MATERIALS AND METHODS: Twenty Arabian horses were tested and 12 were eliminated during the ride and 8 completed the distances. Routine haematological and biochemical tests and measurement of serum concentrations of SAA were carried out before and after the competition. Results were compared using the Mann Whitney U test. RESULTS: Before the competition all haematological and biochemical parameters varied within reference ranges with no differences between the eliminated horses and the ones that successfully finished the competition. After the rides creatine phosphokinase activity and neutrophil: lymphocyte ratio reflecting exercise-induced leukogram changes increased (P < 0.05) in both groups. Before the competition, the concentration of SAA remained within reference ranges; however, it was significantly (P < 0.05) lower in horses that successfully finished the competition than in eliminated ones (411.7 ± 144 ng/ml vs. 5809.5 ± 2242.7 ng/ml). After the ride SAA levels increased (P < 0.05) and were similar in both groups (13,833.8 ± 1354.3 ng/ml and 12,831.2 ± 1317.6 ng/ml). CONCLUSIONS: Serum SAA level was the only laboratory parameter that indicated most (66.6%) of the eliminated horses before entering the competition. None of the horses with SAA level higher than 1000 ng/ml completed the distance. Thus, it may be postulated that serum SAA concentration may indicate a poor status of a horse, resulting in elimination during a competition.

The effect of the 162 km endurance ride on equine peripheral blood neutrophil and lymphocyte functions.

Strenuous exercise is recognized as a stress, which may induce functional immunodeficiency and increase individual susceptibility to infection. It has been shown in equine athletes, that alterations in leukocyte functions occur after moderate and submaximal exertion, however, no data deal with the effect of extreme physical exertion. In this study, we evaluated leukocyte functions (neutrophil oxidative burst and lymphocyte proliferation activity in response to mitogens) in horses following the CEI 3* 162 km endurance ride. Exercise-induced stress was manifested as neutrophilic leukocytosis and lymphopaenia resulting in a significant increase in neutrophil:lymphocyte ratio. The alterations in neutrophil function were expressed as a lower percentage of the cells undergoing oxidative burst. The spontaneous lymphocyte proliferation was very high, however, the cells failed to respond to mitogens. Although a decrease in lymphocyte proliferation in response to mitogens has been reported previously, the pattern determined in our study was unique. It may suggest that during the extreme physical exercise immune cells receive an excessive stimulation from yet undetermined factor(s), which renders them unresponsive to extraneous mitogens. The differences between alterations in leukocyte activities induced by extreme exertion may reflect the exercise type and duration as well as the training status of the horses.

Evaluation of DNA barcoding and identification of new haplomorphs in Canadian deerflies and horseflies.

This paper reports the first tests of the suitability of the standardized mitochondrial cytochrome c oxidase subunit I (COI) barcoding system for the identification of Canadian deerflies and horseflies. Two additional mitochondrial molecular markers were used to determine whether unambiguous species recognition in tabanids can be achieved. Our 332 Canadian tabanid samples yielded 650 sequences from five genera and 42 species. Standard COI barcodes demonstrated a strong A + T bias (mean 68.1%), especially at third codon positions (mean 93.0%). Our preliminary test of this system showed that the standard COI barcode worked well for Canadian Tabanidae: the target DNA can be easily recovered from small amounts of insect tissue and aligned for all tabanid taxa. Each tabanid species possessed distinctive sets of COI haplotypes which discriminated well among species. Average conspecific Kimura two-parameter (K2P) divergence (0.49%) was 12 times lower than the average divergence within species. Both the neighbour-joining and the Bayesian methods produced trees with identical monophyletic species groups. Two species, Chrysops dawsoni Philip and Chrysops montanus Osten Sacken (Diptera: Tabanidae), showed relatively deep intraspecific sequence divergences (∼ 10 times the average) for all three mitochondrial gene regions analysed. We suggest provisional differentiation of Ch. montanus into two haplotypes, namely, Ch. montanus haplomorph 1 and Ch. montanus haplomorph 2, both defined by their molecular sequences and by newly discovered differences in structural features near their ocelli.

Effect of Helicobacter sp. infection on the number of antral gastric endocrine cells in swine.

The aim of the present study was to evaluate the effect of Candidatus Helicobacter suis (CHS) and other Helicobacter sp. different from Candidatus Helicobacter suis (non-Candidatus Helicobacter suis, non-CHS) infection on the number of endocrine G and D cells and G/D cells ratio in antral gastric mucosa in swine. Twenty nine stomachs were obtained from clinical healthy pigs about 6 months old and weighing approximately 100-120 kg after slaughter at abattoir located in central Poland. From each stomach samples of the antral gastric mucosa were taken for histopathology, and PCR examination for presence of Helicobacter genus and Candidatus Helicobacter suis. Samples for histopathology and immunohistochemistry were fixed in 10% buffered formalin. To reveal the expression of gastrin- and somatostatin-producing cells specific antibodies were used. Selected endocrine cells were counted in the midzone of pyloric glands, the results were expressed as a mean of the number of immunoreactive cells in one microscopic field, and as the ratio of gastrin to somatostatin cells (G/D). It can be concluded that some species of swine Helicobacter can alter the number of endocrine cells in gastric antral mucosa. Some of these alterations, for example increase the number of G cells, decrease of the D cells and especially increase of ratio G to D cells can be responsible for development of gastroesophageal ulcers in swine.

The diagnosis of gastritis and helicobacter-like organisms infection in endoscopic biopsies of the canine gastric mucosa.

The aim of this study was to evaluate the prevalence of gastric Helicobacter-like organisms (GHLO) and gastritis in the gastric mucosa of dogs with gastric disorders. Tissue samples of the gastric mucosa were obtained from 30 dogs with gastrointestinal symptoms (vomiting, abdominal pain or discomfort, loss of appetite) during endoscopy. Histopathological examinations were performed and occurrence of GHLO infection, gastritis and other mucosal changes were estimated. The GHLO infection and gastritis were identified in 63.3 and 36.6% of dogs respectively; other mucosal changes included fibrosis in the lamina propria, degenerative changes of the gastric glands and hyperplasia of the parietal cells. The present study has revealed that microscopically found gastritis is not frequent in dogs examined by endoscopy. GHLO infection can be responsible for some cases of gastritis and hyperplasia of parietal cells in dogs.

Immunobiology of bovine mammary gland: apoptosis of somatic cells in milk during naturally occurring mastitis.

Mastitis remains a major cause of economic losses in dairy herds. It is believed, that the enhancement of natural defense mechanisms in mammary gland may be helpful in the treatment of bovine mastitis. Our study was designed to assess the apoptosis of leukocytes isolated from bovine milk during subclinical staphylococcal mastitis. Milk samples were collected from cows naturally infected with one pathogen--Staphylococcus aureus and from animals with mastitis caused by several pathogens, including S. aureus. It has been determined that the rate of apoptosis was lower in mastitic milk, as compared with control samples, although varied significantly between groups. High percentage of apoptotic cells was detected in milk with high counts of pathogenic bacteria. In all groups the rate of apoptosis was dependent on the bacterial load, although various correlations were identified. Thus, it is postulated, that the rate of apoptosis of somatic cells in mastitic milk is related to the etiology of infection and is determined by the bacterial load.

Identifying Canadian mosquito species through DNA barcodes.

A short fragment of mt DNA from the cytochrome c oxidase 1 (CO1) region was used to provide the first CO1 barcodes for 37 species of Canadian mosquitoes (Diptera: Culicidae) from the provinces Ontario and New Brunswick. Sequence variation was analysed in a 617-bp fragment from the 5' end of the CO1 region. Sequences of each mosquito species formed barcode clusters with tight cohesion that were usually clearly distinct from those of allied species. CO1 sequence divergences were, on average, nearly 20 times higher for congeneric species than for members of a species; divergences between congeneric species averaged 10.4% (range 0.2-17.2%), whereas those for conspecific individuals averaged 0.5% (range 0.0-3.9%).

p53 expression in canine lymphoma.

The tumour suppressor p53 plays a key role in DNA damage and repair. It is the most frequently altered gene in human cancers and these mutations may implicate the genesis and/or progression of tumours. Mutations of the p53 gene were also found in a number of canine cancers, although it is poorly estimated in canine lymphomas. Thus, the aim of this study was to investigate the p53 status in these types of tumours. We have shown that the expression of p53 in canine lymphomas is rare, however significantly differs between lymphomas of T- and B-cell origin.

Equine herpes virus 2 infection in horse populations in Poland.

The prevalence of Equine herpesvirus 2 (EHV-2) infections in the horse populations in Poland was investigated. Peripheral blood leukocytes (PBLs) of 139 horses were tested. The animals were divided into four groups: clinically healthy horses, horses suffering from respiratory disorders, mares with a recent abortion and horses with diagnosed ataxia. Thirty-four virus isolates were obtained from leukocytes of the tested animals by cocultivation with equine dermal cells and were identified as EHV-2 by PCR using primers for the gB gene of EHV-2 and/or primers for the sequence located upstream of the gene homologous to the equine interleukin 10 (IL-10) gene. These results indicate that EHV-2 is prevalent in horse populations in Poland. As the virus was most frequently isolated from horses with respiratory disorders its etiological importance may be considered.