Mycobacterium caprae tuberculosis in a captive lion in Ukraine - Case Report.

Introduction and Objective. Because of the armed conflict in Ukraine, companion, farm and captive wild animals have been moved in a simplified procedure across the Polish-Ukrainian border. For that reason, in 2022, Poznan Zoo provided support for almost 200 wild animals before movement to facilities in other countries. The aim of the study is to identify the under-recognized risk of moving animals that may be infected with zoonotic pathogens, such as Mycobacterium caprae, between countries. Materials and Method. Sections of the heart, lungs, and mesentery of 4-year-old lioness from Ukraine were taken for histopathological and bacteriological examination. Results. Microbiological examinations confirmed the presence of Mycobacterium caprae, SB0418 spoligotype. Conclusion. TB is a zoonotic disease present globally. Movement of captive wild animals from regions with MTBC cases, or lack of MTBC surveillance, such as UA may pose a potential threat to public health.

Interleukin-6 as a Milk Marker of Clinical and Subclinical Intramammary Infections (IMI) in Cows Caused by Streptococcus spp.

The aim of the study was to evaluate the concentrations of Interleukin-6 (IL-6) in milk and serum of healthy cows (HE) and cows with mastitis caused by Streptococcus spp. The blood and milk samples were obtained from Holstein-Friesian cows (Lublin region, Poland). A total of 43 milk and serum samples from 28 cows with mastitis and 15 healthy cows were selected for study. IL-6 levels in milk from HE cows ranged from 6.09-80.24 pg/mL (median 26.6 pg/mL) and were significantly lower than in milk from both cows with clinical and subclinical mastitis (487.09 pg/mL vs. 26.6 pg/mL in CM, p < 0.001; and 165.31 pg/mL vs. 26.6 pg/mL in SCM, p < 0.001). The IL-6 concentration in the serum of HE was not significantly different from the serum IL-6 of the entire group of mastitis cows, regardless of whether the inflammation proceeded in a clinical or subclinical form (44.37 pg/mL vs. 78.09 pg/mL; 128.29 pg/mL vs. 78.09 pg/mL, respectively). The present study indicates that cows with mastitis caused by Streptococcus spp. develop a local immune response in the mammary gland in response to the pathogen. Monitoring of IL-6 levels in milk can allow early detection of mastitis, which is especially important in cases of subclinical inflammation.

Cytokine inflammatory response in dairy cows with mastitis caused by Streptococcus agalactiae.

Introduction: The aim of the study was evaluation of the concentrations of interleukin (IL)-1ß, IL-8, IL-12ß and tumour necrosis factor alpha (TNF-α) in the serum and milk of cows with mastitis caused by Streptococcus agalactiae. Material and Methods: A total of 60 milk samples from diseased cows and 30 milk samples from healthy cows were included in the study. Blood and milk samples were taken from Holstein-Friesian cows from three herds (two in tie-stall and one in a free-stall housing system) in Lublin Province in Poland. The concentrations of cytokines in blood serum and quarter milk samples were determined by ELISA. Results: The levels of IL-1ß, IL-8, IL-12ß and TNF-α were significantly higher in the milk of cows suffering from mastitis caused by S. agalactiae compared to the milk of healthy cows (263.03 vs 55.36 pg/mL, 298.34 vs 131.82 pg/mL, 604.10 vs 139.17 pg/mL and 460.86 vs 78.82 pg/mL, respectively). In the group of sick cows, cytokine levels were significantly higher in milk than in serum (263.03 vs 55.25 pg/mL for IL-1ß, 298.34 vs 164.22 pg/mL for IL-8, 604.10 vs 70.34 pg/mL for IL-12ß and 460.86 vs 104.78 pg/mL for TNF-α). Conclusion: The results confirm the involvement of the entire bovine immune system to protect against the bacteria first locally in the udder. The response of the mammary gland to infection caused by S. agalactiae is rapid and already very strong at the beginning of the infection.

Comparative analysis of total protein, casein, lactose, and fat content in milk of cows suffering from subclinical and clinical mastitis caused by Streptococcus spp.

Introduction: The aim of the study was to analyse the total protein (TP), casein (CAS), lactose (LAC), and fat content of milk from cows with subclinical (SCM) and clinical mastitis (CM) caused by Streptococcus spp. Material and Methods: A total of 60 milk samples from diseased cows and 30 milk samples from healthy cows were included in the study. Milk samples were taken from Holstein-Friesian cows from four dairy farms in Lublin Province. The bacteriological examination of the milk was performed and the somatic cells count in 1 mL of milk was determined using a SomaCount FC automatic cell counter. Determination of TP, CAS, LAC, FAT and FA levels in milk was carried out using a DairySpec FT automated Fourier transform infrared spectrometer. Results: Total protein in milk from HE was significantly higher than in milk from cows with mastitis (4.04% vs 3.57% in milk from SCM cows and 3.7% in milk from CM cows, P = 0.001). The CAS level was 2.73% in milk from CM cows and 2.92% in milk from SCM cows vs 3.30% in milk from HE cows, P = 0.001. The changes in CAS and TP in milk resulted in a significant difference in the CAS/TP ratio (81.7% in milk from HE cows vs 73.8% in milk from CM cows). A decrease in levels was also recorded for LAC (4.8% in milk from HE cows vs 4.51% in milk from SCM cows and 4.01% in milk from CM cows, P = 0.001). The fat level was significantly higher in milk from healthy cows than in milk from cows with mastitis (4.0% vs 2.3% in milk from SCM cows and 1.64% in milk from CM cows, P = 0.001). Conclusion: It should be emphasised that the decrease in the levels of TP, LAC and FAT was significant not only in milk from CM cows but also in milk from SCM cows. This is very unfavourable, because the reduction in the main milk components results in poor quality dairy products and impairs line processes.

Divergences between serum C-reactive protein and ferritin concentrations in canine pyometra.

The main aim of this report was to investigate and compare the response of serum C-reactive protein (CRP) and ferritin, two positive acute phase proteins (APPs) which usually show an increase in inflammatory processes, in dogs with pyometra. For this purpose, two different studies were made. In the first one , both proteins were measured together in an APPs profile in 25 dogs with pyometra, 25 dogs with pancreatitis (as an example of a positive inflammatory control group), and in 25 healthy dogs. In the second study, to advance the knowledge of the changes and evolution of serum ferritin and CRP in dogs with pyometra after treatment, the concentrations of both APPs were analyzed in 30 dogs with pyometra at diagnosis and after ovariohysterectomy and in 10 clinically healthy female dogs before and after elective spaying. In both studies, bitches with pyometra showed significant increases in serum CRP, indicating an inflammatory condition, but not in serum ferritin despite being a moderate positive APP. This divergence between the dynamics of these APPs could be a useful tool for the suspicion of cases of canine pyometra.

Assessment of Selected Immunological Parameters in Dairy Cows with Naturally Occurring Mycotoxicosis before and after the Application of a Mycotoxin Deactivator.

Introduction: Mycotoxins in dairy cows can cause many non-specific symptoms often resulting from immune system overreaction. The study assessed the concentration of selected cytokines and acute phase proteins (APP) in cows with natural mycotoxicosis before and after using a mycotoxin neutraliser. The cytokines were tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 10 (IL-10), and the APP were serum amyloid A (SAA) and haptoglobin (Hp). Material and Methods: The research was carried out on an experimental group (Exp) of 10 herdmate Holstein-Friesian cows with mycotoxicosis. The control group (Con) was 10 healthy cows of the same breed from a different herd. Cows in the Exp group were administered the mycotoxin deactivator Mycofix for three months. Blood was drawn from Exp cows once before administering Mycofix and a second time after three months of its use. Blood was also drawn from Con cows at the same times. Serum levels of TNF-α, IL-6, IL-10, SAA and Hp were assessed using ELISA. Results: The concentrations of all cytokines and Hp in Exp cows were higher before treatment (P < 0.001) than those in Con cows. After three months of administering Mycofix, the concentrations of TNF-α and IL-6 were significantly lower than their pre-treatment levels (P < 0.001). The concentrations of IL-6, IL-10, and Hp were still significantly higher than those in the Con group (P < 0.001). In cows with mycotoxicosis, simultaneous stimulation of antagonistic processes was noted: a pro-inflammatory process in the upregulation of TNF-α and IL-6, and an anti-inflammatory one in the upregulation of IL-10. Conclusion: Despite the absorbent's use and the resolution of clinical symptoms in Exp cows, high levels of IL-10 and Hp and IL-6 were maintained. Assessment of the level of cytokines and APP appears to be a useful and precise tool for the evaluation and application of the appropriate dose of the mycotoxin absorbent or the evaluation of its effectiveness.

Activity of the glycosidases ß-galactosidase, α-l-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase in uterine tissues from female dogs in diestrus with and without pyometra.

This study aimed to compare the activity of selected glycosidases (ß-galactosidase, α-l-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase) in homogenates of uterine tissues obtained from female dogs with and without pyometra. In addition, it examined the availability of substrates for these glycosidases in the homogenates. The study was carried out on female dogs undergoing ovariohysterectomy for pyometra (n = 10) and clinically healthy dogs (n = 10) undergoing elective spaying. The activity of ß-galactosidase, α-l-fucosidase, and ß-N-acetyl-hexosaminidase was analyzed using a spectrofluorometer and that of sialidase using a colorimetric method. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis with Alcian Blue (AB) and Periodic Acid-Schiff (PAS) staining was performed to determine the presence of substrates for these glycosidases in the homogenates of uterine tissues. The results revealed that the activity of all the examined glycosidases was significantly higher (P < 0.05) in the uterine tissues isolated from dogs with pyometra in comparison to healthy dogs. The electrophoretic patterns of the selected samples showed several proteins, which contained different sugar moieties stained by AB and PAS and the profiles differed significantly between the pyometra group and the healthy group. Densitometric analysis of AB staining showed patterns between 233 and 148, 86 and 55, and 43 and 20 kDa, which differed markedly in sugar content between the examined groups of animals. Similarly, PAS staining analysis revealed patterns of different molecular weights, between 233 and 117 and between 55 and 32 kDa, which also differed in sugar content. These findings suggest that canine pyometra is accompanied by the increase in the activity of selected glycosidases in the uterus. This could potentially modify the glycan structures of uterine glycoproteins and in result their biological functions. Further studies are needed to elucidate the potential role of the increased activity of glycosidases in the pathogenesis of this disease.

Content of tryptophan and kynurenines in serum and milk of dairy cows with mastitis caused by Streptococcus spp.

The aim of the study was to investigate serum and milk concentrations of tryptophan (TRP), kynurenine (KYN) and kynurenic acid (KYNA), and activity of indoleamine 2,3-dioxygenase (IDO) in cows suffering from mastitis caused by Streptococcus spp. The blood and milk samples were collected from Holstein-Friesian cows farmed in the Lublin region of Poland. It was found that TRP was lower in cows with mastitis both in serum and milk. KYN was lower in serum but not in milk. KYNA was not significantly altered in diseased cows both in serum and milk. The activity of IDO calculated as KYN to TRP ratio was unchanged in serum but was markedly elevated in milk of cows with mastitis. Our findings may have important implications for diagnosis of mastitis in cows because an increase of activity of IDO and reduction of TRP in milk might be a valuable early marker predicting the occurrence of the disease.

Tryptophan, Kynurenine and Kynurenic Acid Concentrations in Milk and Serum of Dairy Cows with Prototheca Mastitis.

The aim of this work was to investigate serum and milk levels of tryptophan (TRP), kynurenine (KYN), and kynurenic acid (KYNA), as well as the activity of indoleamine 2,3-dioxygenase (IDO) in cows with mastitis due to Prototheca algae. The study was prompted by previous research showing a link between the KYN pathway of TRP metabolism and bovine mastitis of bacterial etiology. The study was carried out over a 2-year period (2018-2019) and included quarter milk and serum samples collected from six dairy herds in Poland. The samples were obtained from healthy cows and cows with Prototheca mastitis of either clinical and subclinical manifestation, as determined upon direct measurement of the somatic cell count or indirectly by performing a California Mastitis Test on suspected quarters. Both TRP and KYN concentrations were significantly lower in milk of mastitic cows compared to healthy animals (0.8 vs. 8.72 µM, p = 0.001; 0.07 vs. 0.32 µM, p = 0.001, respectively). The difference in TRP and KYN concentrations in the sera of the two animal groups was much less pronounced (25.55 vs. 27.57 µM, 3.03 vs. 3.56 nM, respectively). The concentration of KYNA was almost at the same level in milk (1.73 vs. 1.70 nM) and in serum (80.47 vs. 75.48 nM) of both mastitic and healthy cows. The data showed that the level of TRP and its metabolites in serum was conspicuously higher compared to milk in all cows under the study. The activity of IDO was significantly higher in milk of cows with Prototheca mastitis compared to healthy animals (71.4 vs. 40.86, p < 0.05), while in serum it was pretty much the same (135.94 vs. 124.98, p > 0.05). The IDO activity differed significantly between serum and milk both for mastitic (135.94 vs. 71.4, p < 0.05) and healthy cows (124.98 vs. 40.86, p < 0.001). In conclusion, low values of TRP and KYN concentrations or elevated IDO activity in milk samples might be used as markers of mastitis due to infectious causes, including Prototheca spp.

Aerobic Isolates from Gestational and Non-Gestational Lactating Bitches (Canis lupus familiaris).

Mastitis is a complex and well-defined mammary gland pathology, and an emergency in bitches. In dogs, its prevalence is about 1% of all reported diseases and about 5.3% of all reproductive pathologies. Lactating bitches are naturally prone to developing mastitis since puppies can easily overstimulate the epidermal layer of nipples during feeding, facilitating bacterial colonization of the glands. This study aimed to describe the aerobic bacterial flora isolated from milk samples derived from a cohort of patients (n = 87) diagnosed with clinical mastitis (n = 29), subclinical mastitis (n = 17) and healthy mammary glands (n = 46). All of the patients underwent a gynecology consultation to diagnose mammary gland afflictions; physical examination results were coupled with traditional hematological findings. The milk samples were plated on specific microbiological media for bacterial isolation. Among the 162 milk samples analyzed, 93.2% (151/162) had a positive microbiological result, while 6.8% (11/162) were sterile. The bacteriological profile of the milk samples showed 47 different species. The most common bacterial families detected in healthy bitches and bitches with subclinical and clinical mastitis were the Staphylococcaceae, Enterobacteriaceae and Enterococcaceae families. The results indicated that half of the isolated bacteria are novel findings in dogs and that some of them are normal components of human milk.

MicroRNA and vascular endothelial growth factor (VEGF) as new useful markers in the diagnosis of benign prostatic hyperplasia in dogs.

Numerous specific biomarkers with a prognostic and diagnostic value comparable to histopathological findings are now used for non-invasive diagnosis of prostate diseases in humans. Meanwhile, as far as dogs are concerned, the diagnosis of prostate disorders is based solely on clinical examination and ultrasound (USG). Therefore, the aim of the study was to assess the usefulness of two biomarkers, i.e. miRNA-129 and VEGF for the diagnosis of BPH in dogs. The study involved 40 dogs divided into three groups. Group I (n = 9) comprised healthy dogs up to the age of 5 years, Group II (n = 17) comprised dogs between the ages of 5-10 suffering from BPH as confirmed by the examination and Group III (n = 14) comprising dogs over 10 years of age, which also had BPH confirmed. The results demonstrated that dogs in group II and III exhibited a significant decrease in miRNA expression (P < 0.0001) and a significant increase in serum VEGF levels (P = 0.025) when compared to the dogs in group I. There was also a positive correlation between the prostate size and VEGF level. The findings led to the conclusion that the determination of miRNA-129 and VEGF can significantly contribute to the diagnosis of prostate disorders in dogs.

Measurement of ITIH4 and Hp levels in bitches with pyometra using newly developed ELISA methods.

Measurement of acute phase proteins (APPs) as biomarkers in canine medicine is in increasing demand. In the present study, the development and validation of two ELISA methods for the quantification of canine inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) are shown. The adequate imprecision and accuracy and wide analytical range make the developed methods appropriate to quantify ITIH4 and Hp in serum samples. The inter- and intra-assay CVs were lower than 10 %, and the assays maintained linearity under dilution and showed analytical equivalence with the method of radial immunodiffusion. The measurement of CRP, Hp and ITIH4 in sera from bitches affected by pyometra allowed us to determine that ITIH4 behaves as a moderate APP in dogs. The group of bitches affected by pyometra showed very high levels of CRP (147 ± 91 mg/L), corresponding to a strong inflammatory process, which resulted in a moderate increase in the concentrations of Hp (7 times) and ITIH4 (3 times) compared to the control group.

Changes in interferon-gamma and neopterin in female dogs undergoing ovariohysterectomy as elective spay or as treatment of pyometra.

The objective of this study was to determine serum concentrations of interferon-gamma (INF-γ) and neopterin (Np) in dogs with pyometra admitted for surgical treatment and to compare these concentrations to healthy dogs admitted for elective spay. The effects of the surgical procedure were also evaluated by measuring these markers in both groups of dogs before and after ovariohysterectomy. Our study indicates that pre-surgery concentrations of INF-γ (57.4 ± 26.0 pg/mL) and Np (5.6 ± 0.8 nmol/L) in healthy dogs were significantly lower compared to dogs with pyometra (124.3 ± 87.6 pg/mL for INF-γ; 7.0 ± 1.5 nmol/L for Np) (P < 0.05 in both cases). Furthermore, Np was lower in dogs with pyometra 3 days after surgery compared to healthy controls (P < 0.001). During the post-operative period, INF-γ showed no statistically significant changes in any of the groups, while Np showed lower serum concentration on day 3 than on day 0 in the pyometra group (P < 0.001). No statistically significant correlation was detected between serum concentrations of INF-γ and Np. These results indicate that pyometra causes alterations in serum concentrations of INF-γ and Np in female dogs compared to physiological levels before surgery and during the postoperative period.

L'objectif de la présente étude était de déterminer les concentrations sériques d'interféron gamma (INF-γ) et de néoptérine (Np) chez des chiens avec pyomètre admis pour traitement chirurgical et de comparer ces concentrations à celles de chiens en santé admis pour stérilisation élective. Les effets de la procédure chirurgicale furent également évalués en mesurant ces marqueurs dans les deux groupes de chiens avant et après ovariohystérectomie. Notre étude indique que les concentrations pré-chirurgie d'IFN-γ (57,4 ± 26,0 pg/mL) et de NP (5,6 ± 0,8 nmol/L) chez les chiens en santé étaient significativement inférieures comparativement aux chiens avec pyomètre (124,3 ± 87,6 pg/mL pour INF-γ; 7,0 ± 1,5 nmol/L pour Np) (P < 0,05 dans les deux cas). De plus, Np était plus bas chez les chiens avec pyomètre trois jours après la chirurgie comparativement aux témoins en santé (P < 0,001). Durant la période post-chirurgicale, INF-γ ne montra aucun changement statistiquement significatif dans aucun des trois groupes, alors que Np a présenté des concentrations sériques plus faibles au jour 3 qu'au jour 0 dans le groupe avec pyomètre (P < 0,001). Aucune corrélation statistiquement significative ne fut détectée entre les concentrations sériques d'INF-γ et de Np. Ces résultats indiquent que le pyomètre cause une altération des concentrations sériques d'INF-γ et de Np chez les chiennes comparativement aux niveaux physiologiques avant la chirurgie et durant la période post-opératoire.(Traduit par Docteur Serge Messier).

Serum amyloid A as an marker of cowÖ¨ s mastitis caused by Streptococcus sp.

The aim of the study was to evaluate the concentrations of amyloid A in serum (SAA) and in milk (MAA) of cows with mastitis caused by Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis and healthy cows. The blood and milk samples were obtained from Holstein-Friesian cows with clinical signs of mastitis from two tie-stall housing systems herds in the Lublin region in Poland. A total of 80 milk and serum samples from 30 cows with mastitis and 10 healthy cows were selected for study. In the quarter milk samples from cows with mastitis Streptococcus strains were isolated: Strep. agalactiae (7 cows), Strep. dysgalactiae (9 cows) and Strep. uberis (14 cows). The present study indicates that amyloid A concentration was significantly higher in milk of cows with mastitis compared to control cows (1134.25 ng/mL and 324.50 ng/mL, P < 0.001). The highest concentration of amyloid A was found in milk of cows with mastitis caused by Strep. agalactiae and Strep. uberis whereas lowest in the milk of cows with mastitis caused by Strep. dysgalactiae (3882.50 ng/mL, 2587.75 ng/mL and 812.00 ng/mL, respectively). No statistically significant difference in amyloid A concentration in serum was revealed between all unhealthy cows and control group (2140.00 ng/mL and 2510.00 ng/mL, P > 0.05). There was also no statistically significant difference between the level of amyloid A in serum and in milk of cows with mastitis caused by Strep. agalactiae and Strep. uberis. Whereas, in the case of Strep. dysgalactiae, like in the group of healthy cows, the level of amyloid A was significantly higher in serum compared to this in milk (2100 ng/mL and 812.00 ng/mL, P < 0.001; 2510.00 ng/mL and 324.50 ng/mL, P < 0.001; respectively).

Selected leukocyte subpopulations in peripheral blood and uterine washings in cows before and after intrauterine administration of cefapirin and methisoprinol.

The aim of the study was to evaluate the selected lymphocyte subpopulations TCD4, TCD8, BCD21, BCD25, CD18, CD11b, and MHC II in blood and uterine flush of cows with endometritis, before and after intrauterine (i.u.) administration of cefapirin and methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. preparation used: Group A, cefapirin; Group B, methisoprinol; Group C, cefapirin and methisoprinol simultaneously; and a control group-without medication. The study was performed using flow cytometry method. Summarizing the results of the research, i.u. infusion of cefapirin caused a weakening of the effector phase of the local uterine immune response; however, it enhanced leukocyte chemotaxis and antigen presentation. After i.u. administration of methisoprinol, the stimulation of specific uterine immunity mechanisms was mainly observed. The use of both mentioned preparations showed the strengthening of specific uterine immunological mechanisms presumably caused by methisoprinol, despite the inhibitory effect of the antibiotic. Intrauterine use of immunostimulatory substances can improve the effectiveness of the endometritis treatment in cows by improving specific local mechanisms of uterine immunity. As a consequence, it may enhance the effector function of immune competent cells and finally eliminate inflammation.

Decorin concentrations in canine normal and neoplastic mammary gland tissues.

In the present study, the concentration of decorin in canine normal and neoplastic mammary gland tissues was examined to understand the potential role of decorin in development and progression of canine mammary tumours. The homogenates of 48 mammary gland tumours (10 benign and 38 malignant) and 10 samples of normal canine mammary gland tissue were used in the study. The presence and quantification of decorin was examined in the homogenates using Western blot and specific canine ELISA. Western blotting confirmed the presence of decorin both in the normal mammary gland tissues and in the mammary gland tumours. The concentration of decorin was significantly higher (p < .05) in the benign tumours and non-metastatic malignant tumours than in the normal mammary gland. The concentration of decorin was significantly lower (p < .05) in the malignant tumours with metastasis to regional lymph nodes compared with benign tumours and non-metastatic malignant tumours. No significant differences were found in the level of decorin between the benign and the non-metastatic malignant tumours. Both the histological type of malignant tumours and the histological grade did not significantly affect the concentration of decorin. These findings suggest that neoplastic transformation in the canine mammary gland leads to increase in the decorin protein synthesis. The reducing decorin concentration in canine malignant mammary tumours appears to facilitate the metastatic spread of these tumours.

The effect of pyometra on glycosylation of proteins in the uterine tissues from female dogs.

The main aim of this study was to investigate the effect of pyometra on glycosylation of proteins in the uterine tissues from female dogs, using western blotting with selected lectins (Sambucus nigra agglutinin - SNA and Maackia amurensis agglutinin - MAL II). In addition protein pattern of examined tissues was also evaluated. The study was performed on 10 female dogs undergoing ovariohysterectomy because of pyometra and 10 clinically healthy female dogs, undergoing elective spaying (ovariohysterectomy). Uterine tissue samples of 1 cm2 were taken from the middle region of each uterine horn in both group of animals immediately after ovariohysterectomy. Tissue samples were homogenized and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting with SNA and MAL II. SDS-PAGE analysis showed differences between pyometra samples and controls in the amount of obtained protein fractions and the protein content in the individual fractions. Five protein (with a molecular weight of 193.78 kDa, 103.18 kDa, 77.67 kDa, 70.39 kDa, and 53.00 kDa) were found only in the pyometra samples. The remaining fractions differed in intensity of staining, which indicated differ abundance of a given protein. The results of western blotting with SNA and MAL II demonstrated that the pattern obtained from densitometric analysis differs between adequate healthy and pyometra samples with regard to the amount of protein fraction obtained as well as the intensity of staining of particular fraction. The pyometra tissues contained seven SNA-binding proteins (with a molecular weight 189.94 kDa, 165.51 kDa, 100.94 kDa, 59.42 KDa, 41.32 kDa, 35.16 kDa, and 32.6 kDa) that were not in the healthy tissues. Of the nine remaining fractions, six showed significantly higher (P < 0.05) intensity of staining in the healthy uterine tissues. In turn, the MAL II-binding protein with a molecular weight 75.85 kDa, 51.12 kDa, and 49.98 kDa were found only in the pyometra samples. Of the 28 remaining fractions, ten demonstrated significantly higher (P < 0.05), and five fractions had significantly lower (P < 0.05) intensity of staining in the pyometra tissues. The results obtained indicate that proteins in uterine tissues from female dogs with pyometra are differently glycosylated compared to normal uterine tissues. These findings provide the basis for further studies of the possible role of glycosylation in the pathogenesis of canine pyometra.

Prevalence of Prototheca spp. on dairy farms in Poland - a cross-country study.

The Prototheca algae have recently emerged as an important cause of bovine mastitis globally. Here, we present results of a first large-scale, cross-country survey on the prevalence of Prototheca spp. in dairy cows, and their environment in Poland. A total of 1211 samples were collected and microbiologically analysed. Included within this number were milk (n = 638), body swabs (n = 374) and environmental samples (n = 199), originating from 400 dairy cows and their surroundings, on 16 dairy farms, based in all major provinces of the country. Prototheca spp. were the third, after Streptococcus and Staphylococcus spp., most common mastitis pathogens. The overall prevalence of protothecal mastitis was 8.3% (33/400), with the majority (75.8%) of cases having a subclinical course, and all but one attributable to P. zopfii genotype 2. Prototheca spp. were cultured from body swabs of both healthy and mastitic cows, yet the isolation rate among the latter was conspicuously lower (12.3% vs. 17.8%). Forty-two (21.2%) environmental samples yielded growth of Prototheca spp. However, no clear association between Prototheca mastitis in dairy cows and the algal isolation from the herd environment was found. Nor was there any association between the environmental recovery of the algae and farm management practices.

A survey on the incidence of Prototheca mastitis in dairy herds in Lublin province, Poland.

Prototheca mastitis has recently become an emerging disease; although its incidence is increasing steadily, its epidemiology remains largely understudied. The aim of this work was to investigate the prevalence of Prototheca spp. in dairy cows and their environment in Lublin province, covering most of southeastern Poland. Between December 2015 and July 2016, a total of 172 milking cows from 10 dairy farms were inspected for mastitis using clinical examination and the California Mastitis Test (CMT). Quarter milk samples (QMS, n = 179) and body site swabs (n = 151) from CMT-positive cows were collected for microbiological culture. In addition, we evaluated QMS and body site swabs from 23 healthy cows, along with 91 environmental samples. Of 100 CMT-positive cows, 71 had at least one QMS positive for microbial growth. In 8 (11.3%) of these cows, originating from 7 dairy farms, Prototheca spp. were cultured. The average somatic cell count of the Prototheca-containing milk was 4.02 × 106 cells/mL compared with 0.13 × 106 cells/mL of the Prototheca-free milk (collected from control animals). No significant differences were observed between mastitis and control cows with respect to counts of total white blood cells, lymphocytes, neutrophils, and eosinophils. Half of the cows with Prototheca spp. in their milk did not yield the algae from other anatomical sites. Eight cows were negative for the presence of Prototheca spp. in their milk but positive for the algae in swabs from anatomical sites. Among the environmental sources that were positive for Prototheca growth were watering troughs, manure, feed, and mud. All (45) Prototheca isolates recovered in this study were subjected to species- and genotype-level molecular identification. All QMS and most of the animal swabs (90%) yielded Prototheca zopfii genotype (gen.) 2. Of the animal samples, P. zopfii gen. 1 and Prototheca blaschkeae were isolated only from feces and rectum. Environmental samples grew either P. zopfii gen. 2 (67%) or P. zopfii gen. 1 (33%). This study demonstrates that P. zopfii gen. 2 is the third most common pathogen of mastitis in cattle in southeast Poland, with an overall incidence of 4.6%. Finding Prototheca spp., including P. zopfii gen. 1 and 2 and P. blaschkeae, in stool and rectal swabs from healthy animals may suggest their role as nonpathogenic microflora of bovine gut.