This study investigated changes in the real-time measured levels of milk β-hydroxybutyrate according milk yield, lactation number and status of reproduction in dairy cows. A total of 378 cows were selected. According to their reproductive status the cows were classified as belonging to the following groups: Fresh (1 – 44 days after calving. n=43). Open (45 – 65 days after calving. n=78), Inseminated (1 – 35 days after insemination. n=133). Pregnant (35 – 60 days after insemination and pregnant (relatively pregnant) (n=124). The cows were milked with DeLaval milking robot (DeLaval Inc., Tumba, Sweden) in combination with a Herd Navigator (Lattec I/S. Hillerød. Denmark) analyser. We observed that milk β-hydroxybutyrate (BHB) had a tendency of increasing with an increase of lactation number. The average BHB in multiparous cows was 11.111% higher in comparison with primiparous cows (p<0.001). We found higher BHB concentration in the multiparous cows in all reproduction status groups (p<0.001). A strong positive statistically significant (p<0.001) relationship has been found between BHB and the average milk yield within all groups of primiparous cows although we found a statistically unreliable coefficient of correlation (from -0.202 to 0.057) between highest milk yield and BHB in primiparous and multiparous cows.
Sperm-mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind exoge- nous DNA and transfer it into oocytes by fertilization. However, SMGT is still undergoing opti- mization to improve its efficiency to produce transgenic animals. The acrosome reaction is neces- sary for spermatozoa to carry the exogenous DNA into oocytes. In this study, the effect of the acrosome reaction on the efficiency of spermatozoa carrying exogenous DNA was evalua- ted. The results showed that the efficiency of the acrosome reaction was significantly higher (p<0.05) after incubation with 50 μmol/L progesterone compared to incubation without proges- terone. It was significantly higher (p<0.05) in the 20, 40, and 60 min of progesterone treatment groups than in the 0 min treatment group. The spermatozoa were further incubated with cyanine dye Cy5 labeled DNA (Cy5-DNA) for 30 min at 37°C, and positive fluorescence signals were detected after the acrosome reaction was induced by progesterone at concentrations of 0 and 50 μmol/L for 40 min. The percentage of positive Cy5-DNA signals in spermatozoa was 96.61±2.06% and 97.51±2.03% following exposure to 0 and 50 μmol/L progesterone, respective- ly. The percentage of partial spermatozoa heads observed following combination with Cy5-DNA was 39.73±3.03% and 56.88±3.12% following exposure to 0 and 50 μmol/L progesterone, respec- tively. The ratio of positively stained spermatozoa combined with exogenous DNA showed no reduction after the acrosome reaction. These results suggest that the acrosome reaction might not be the key factor affecting the efficiency of SMGT.
In the present study a severe outbreak of hemorrhagic pneumonia (HP) in neonatal minks concomitant with Leismania infantum (L. infantum) detection is reported. The outbreak took place on a Greek mink farm and affected 1,362 mink kits, with 524 dying. Macroscopic lesions of 14 necropsied affected kits were confined to the respiratory system with dark red, consolidated lung lobes and to the small intestine with severe, acute, hemorrhagic and necrotic enteritis. Microscopic examination of lung sections revealed severe hemorrhagic pyogranulomatous pneumonia. Bacteria were obtained in pure culture from the lungs of all necropsied animals and were confirmed as Pseudomonas aeruginosa (P. aeruginosa). Three out of 14 (21.4%) animals were positive for the presence of L. infantum DNA. The outbreak was attributed to the infection of minks with P. aeruginosa, possibly as a consequence of being immuno-suppressed by L. infantum. Further research is necessary, especially on the pathogenesis of P. aeruginosa/L. infantum co-infection and the implications of this interaction on HP disease outcome.
The aim of this study was to measure the NO level in boar semen held in a liquid state and to determine its putative relation to spermatozoa motility, plasma membrane integrity, mitochondrial membrane potential and ATP content. Generally, the percentage of spermatozoa which generated nitric oxide gradually increased, while NO level in the surrounding medium declined during the liquid preservation. NO generation in semen preserved in BTS was higher as compared to those in Androhep®Plus. We demonstrated the positive correlation between the NO level in fresh spermatozoa and their quality. We also showed negative correlation between nitric oxide level in spermatozoa preserved in BTS and sperm cells motility as well as plasma membrane integrity. Results obtained in this study confirm that NO may affect sperm physiology in a dualistic manner.
Taking bacterial virulence factors as targets is a new therapy for treating host bacterial infection. The aim of this study was to investigate the effect of matrine on α-hemolysin production of Staphylococcus aureus (S. aureus) and reducing the damage to bovine mammary epithelial cells (BMECs) induced by S. aureus α-hemolysin. Subinhibitory concentrations of matrine decreased the production of α-hemolysin in none dose-dependent manner and matrine exhibited a protective effect on S. aureus-induced BMECs injury. The results indicated that the structure of matrine may potentially be used as a basic structure for development of drugs aimed at curing and preventing dairy bovine mastitis.
Previous studies have compared the effectiveness of dinoprost and cloprostenol in cows yielding conflicting results. The aim of our study was to evaluate the efficacy of single treatment with cloprostenol or dinoprost on estrus and reproductive performance in cows with unobserved estrus after service. The study was conducted over four years in two dairy herds of Polish Holstein Frisian cows under a herd health program with an average milk yield per cow over 9000 L. Cows (n=523) diagnosed ultrasonographically as non-pregnant and with a corpus luteum were randomly assigned to be treated with either cloprostenol (n=261) or dinoprost (n=262). The estrus detection rates after administration of cloprostenol or dinoprost were 59.4%, and 57.6%, respectively. The difference between both groups was not statistically significant (p>0.05). Distribution of observed estrus did not differ between cloprostenol and dinoprost. There were no differences (p>0.05) between cloprostenol and dinoprost in conception rate (65.2% vs. 66.2%, respectively) and pregnancy rate (57.5% vs. 54.9%, respectively). Mean days open were similar in cows of both treatments (177.5 ± 74.6 days vs. 175.8 ± 62.6 days, respectively; p>0.05). In conclusion, data from this study showed no significant differences in estrus detection rates and fertility between cows with unobserved estrus after service treated with cloprostenol or dinoprost. Both products are equally useful for the treatment of non-pregnant dairy cows with anestrus after service within a reproductive herd health program.
Calretinin (CR), a calcium-binding protein from EF-hand family, is localised in non-pyramidal GABA-ergic interneurons of the hippocampus. CR takes part in maintaining calcium binding homeostasis, which suggests its neuroprotective role. Hippocampal neurons contain membrane transient receptor potential vanilloid 1 (TRPV1) which binds to capsaicin (CAP) contained in habanero pepper fruits. Few in vivo studies have revealed the effect of CAP on interneurons containing CR. The aim of the present study was to investigate the CR immunoreac- tivity in interneurons of the hippocampal CA1 field and dentate gyrus (DG) in adult rats after intragastric admin- istration of the habanero pepper fruits. Wistar rats received a peanut oil – control group (C), and oil suspension of habanero pepper fruits at doses of 0.025 g dm/kg b.w. – group I and 0.08 g dm/kg b.w. – group II for 28 days. After euthanasia, the brains were collected and embedded in paraffin blocks using a routine histological tech- nique. Frontal hippocampal sections were immunohistochemically stained for CR by using a peroxidase-antiper- oxidase method. CR immunoreactive (CR-IR) interneurons were morphologically and morphometrically ana- lyzed under a light microscope. The results showed similar shapes and distribution of cells in both areas of the brain in group C and I of animals. However, CR-IR interneurons in the hippocampal CA1 field and in DG were occasionally observed in the group II of rats. The results of morphometric studies did not reveal statistically significant differences in the surface area and shape index of cells between examined brain regions from groups I and II compared to group C. Only in group II of rats, an increase in the digital immunostaining intensity of CR-IR interneurons was found in DG. Low number of CR-IR interneurons in the hippocampal CA1 field and in the DG, under the influence of a large dose of habanero pepper fruits containing CAP, may be caused by the activation of TRPV1 receptors and the increase in Ca2+ ions in these cells. This phenomenon may ultimately lead to neuronal death and may disturb neuronal conduction.
The bacterial species Anaplasma phagocytophilum and Rickettsia helvetica are pathogenic for humans and domestic animals and are transmitted by ticks, e.g., of the Ixodes genus. Most of the vertebrate species constituting reservoirs for anaplasmas are known, but the potential reservoirs of rickettsiae are still under discussion. This study presents an analysis of the DNA of tick-borne pathogens isolated from the whole blood of goats grazing on meadows in West Pomerania, Poland. No DNA of A. phagocytophilum was found in the blood of the goats, while the DNA of R. helvetica was detected in 5.5% of the animals. The potential role of ruminants in the circulation of R. helvetica remains unknown.
There is no information available about the incidence of anestrus and its clinical forms after service in dairy cows in Poland. Thus, the aim of this study was to investigate the incidence of clinical forms of anoestrus after unsuccessful artificial insemination in dairy cows based on ultrasound examination. The study was carried out on 1543 Polish Holstein-Friesian cows in 8 dairy herds in north-east Poland over a three-year period. Cows were examined for pregnancy on day 35 after AI using a Honda 1500 portable ultrasound scanner equipped with a 5 MHz linear-array transducer. Cows diagnosed as pregnant were re-examined on day 45. Of the 1543 inseminated cows, 408 (26.4%) showed no estrus signs and were diagnosed not-pregnant by ultrasonography, 328 (21.3%) returned to estrus within 35 days, and 807 (52.3%) were pregnant via artificial insemination. The incidence of anestrus after service in non-pregnant cows varied among herds from 10.3% to 32.9% of cows (p<0.05). Based on ultrasound examination silent heat was diagnosed in 324 (79.4%), corpus luteum pseudograviditatis in 36 (8.8%), ovarian cysts in 26 (6.4%), and ovarian afunction in 22 (5.4%) of 408 anestrous, non-pregnant cows. The results of this study showed that the incidence of anestrus after service in dairy herds in North-East Poland was high. The most prevalent clinical form of post-service anestrus was silent heat.
Recent outbreaks of adenovirus (FAdV) infections in poultry flocks have been determined in many countries in Europe, Asia and Australia connected with economic consequences, and loses in poultry production. To better understand the evolution and transmission of FAdV viruses, de- tailed codon usage analysis was performed for 137 recently obtained FAdV strains. A high effec- tive number of codons, and an indication the presence of low codon usage were determined. The presence of mutations, and their influence on codon usage was confirmed by a correlation be- tween nucleotide compositions at the 3rd codon positions, HVRs1-4, and ENCs. This presence indicate some influence of natural selection, and antigenic properties of examined FAdV strains.
The objective of this study was to investigate the anticoccidial effect of apple cider vinegar added to drinking water with the anticoccidial effect of amprolium to feed broiler chicken. The study has adopted an observational approach to evaluate the anticoccidial effect of apple cider vinegar on broiler chicken. The antioxidative changes were measured adding natural apple cider vinegar to drinking water. Four hundred and fifty broiler chickens were purchased from the local market and distributed into three groups (T+vc: positive control, T-vc: negative control Tv: apple cider vinegar) with 150 chickens in each group. The three groups were further replicated into 3 blocks each containing 50 chickens. The groups were fed balanced diet, amprolium was added to the feed of positive control group, and apple cider vinegar was added to the water of Tv group. Measurements of the different variables were started from week 3, at the end of each week 3 birds were chosen randomly, blood samples were collected via the wing vein, and fecal oocysts were counted from intestinal contents of each individual bird using the McMaster technique. Broiler in the control groups T+ve and T-ve showed clinical signs of coccidiosis (blood in feces) and the number of coccidial oocytes in feces increased with time. In the vinegar group, no clinical signs of coccidiosis were observed. Concentrations of total antioxidants and catalase enzyme activity significantly increased (p≤0.05); while malondialdehyde concentration significantly decreased (p≤0.05).
In this study the potential usefulness of infrared thermography (IRT) as a non-invasive tool to rapidly screen the most common non-infectious foot lesions in dairy cows was evaluated. Thirty-eight healthy cows and 38 cows affected by foot diseases were enrolled. Diseased cows showed the following disorders at lateral and medial claw in the hind foot: white line lesion, sole ulcer, sole haemorrhage, horizontal fissure, axial fissure. Thermography images of hind foot were collected for each animal using a digital infrared camera. Foot temperature was measured in four regions: central area of the hind foot (A1), interdigital area of the hind foot (A2), lateral (A3) and medial (A4) claw in the hind foot. Higher temperature values in the regions A1 and A2 compared to A3 and A4 were found in both healthy and diseased cows (p0.001). Cows affected by foot diseases showed higher foot temperature values compared to healthy cows (p0.05) in all considered regions. This study highlights the potential application of IRT as a reliable, practical tool for detection of hoof lesions in dairy cows. Multiple scanning images and comparisons between affected and healthy anatomical structures could be useful in defining the consistency of abnormality.
Inflammation together with lipolysis and ketogenesis in early lactation can cause low productivity and may be harmful to the cow health. The objective of the study was to determine if ketoprofen treatment in the first days following parturition would positively affect the milk production and whether it was associated with the metabolic and inflammatory response. The study was conducted on 30 cows divided into two groups of 15 cows each. The experimental group was treated with 3 mg × kg. bw. -1 ketoprofen for three consecutive days after parturition. The blood samples were collected on the first day of treatment and in the first and second week postpartum and they were analyzed for non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), tumour necrosis factor-a(TNF-a) and haptoglobin. The results suggested that ketoprofen-treated cows with a higher milk production had a significantly lower concentration of NEFA, BHB, TNF-a and haptoglobin in the first and second week postpartum. No differences were found in the control group in metabolic status regardless of the achieved level of milk production. Ketoprofen administration in postpartum cows can enhance the milk yield. The higher milk yield in the experimental group might be associated with a lower degree of lipolysis, ketogenesis and reduced inflammatory response in the first two weeks postpartum.
One of the most common reasons for horse lameness is subchondral bone cysts (SBCs), which are especially evident in young horse athletes. It is believed that SBC development is strongly associated with an individual’s bone growth and/or bone microstructure impairment. Current methods of SBC treatment include pharmacological treatment or surgical procedures which may allow the bone within the cyst to rebuild and be restored to properly developed bone tissue. Thus, we propose filling the SBCs with a 3D complex of alginate hydrogel and autologous adipose derived mesenchymal stem cells (ASCs). We have observed at the in vitro level, that this hydrogel complex induces osteogenic and chondrogenic differentiation potential through the upregulation of bone morphogenetic protein, osteopontin, collagen type I and aggrecan mRNA levels. Moreover, we detected the creation of a 3D extracellular matrix (EM). To investigate the complex in vivo, we chose 8 horses of varying age suffering from SBC, which resulted in lameness, to undergo experimental surgery. We documented the horses’ clinical appearance, lameness and radiographic appearance, to determine that there was clinical improvement in 87.75% of the patients (n=7, out of 8 horses) 6 months postoperatively and 100% (n=8, out of 8 horses) a year after surgery. These results are promising for the potential of this procedure to become the standard in SBC treatment.
Current study was designed to investigate the protective effects of royal jelly on Flunixin me- glumine (FM)-induced spermiotoxicity related to sperm concentration, abnormal spermatozoa count and histopathological changes in mice testis. The subjects were divided into five groups according to FM and/or royal jelly intake: Control group; group 1, FM alone (25 mg/kg, im); group 2, combination of FM (25 mg/kg, im) and royal jelly (200 mg/kg, oral); group 3, FM alone (50 mg/kg, im); and group 4, combination of FM (50 mg/kg, im) and royal jelly (200 mg/kg, oral). The animals were fed once daily for 15 days and they were sacrificed last day. Epididymal sperm concentration and abnormal spermatozoa count were noted. Testicular histological findings were evaluated. On purpose, organization of each animal was graded according to Johnsen’s scoring to assess the spermatogenesis relying on seminiferous tubule cross-section scores. Comparing to controls, FM administration caused a decrease in sperm concentration (p<0.05), an increase in total abnormal spermatozoa rates (p<0.05) and more degenerative changes in testes in mice. Royal jelly supplementation ameliorated both sperm concentration and abnormal spermato- zoa (p<0.05) comparing to the control group. In conclusion, we suggested that royal jelly might have protective effects in the FM-induced reductions in epididymal sperm concentration and in- crease in abnormal spermatozoa rate.
Despite the consensus on the role of lung and pleura ultrasound in human medicine, veteri- nary medicine questions credibility of the pulmonary evaluation in ultrasound examination, based on the analysis of artifacts in animals with clinical signs of respiratory failure and possibility of pulmonary edema diagnosis with recognition of the degree of its severity. The study was conduct- ed on 47 animals (29 dogs and 18 cats) of different breeds, age and sex. In all of animals prior to the transthoracic lung and pleura ultrasound examination (TLPUS), all animals were subjected to a clinical examination and hematological blood test as well as chest radiography examination in three projections. Ultrasound imaging of the chest in each animal was performed at designated four defined segments. TLPUS in dogs and cats based on an analysis of artifacts allows recogni- tion of pulmonary edema, to the degree comparable to chest X-ray examination. The number of depicted B-lines artifacts is proportional to the degree of pulmonary edema. These results allow to reduce the number of radiographs and allow the shortening of the diagnostic process for pa- tients in life-threatening condition.
In humans, iron deficiency represents a relevant occurrence in heart failure (HF), with or without anaemia, and is associated with the worst outcome. Moreover, chronic kidney disease (CKD) is a well-known comorbidity of HF and is strongly associated with the risk of developing anaemia. The most common cause of HF in dogs is myxomatous mitral valve disease (MMVD). To the best of our knowledge, no studies have examined the iron status in dogs with HF, with and without CKD. The aim of this retrospective study was to evaluate the iron status in dogs affected by MMVD and how strong is the relation with HF. The retrospective study included 54 dogs with complete case records, echocardiography and laboratory analyses. Iron status was evaluated by measuring serum iron concentration (SIC), un- saturated iron binding capacity (UIBC), total iron binding capacity (TIBC), and percentage of saturation (%SAT). The prevalence of dogs showing low serum iron concentration (SIC) was 18% in the whole population, 33% in symptomatic patients, 100% in dogs with acute decompensated HF. No signif- icant differences in SIC, UIBC, TIBC and %SAT median values were found among dogs classi- fied in different ACVIM (American College of Veterinary Internal Medicine) classes, between symptomatic and non-symptomatic patients, and among IRIS (International Renal Interest Soci- ety) classes. Azotemic and non-azotemic patients presented a significant difference in SIC mean values (p=0.02). Generalised linear model (GLM) revealed that dogs with low SIC were at high- er risk of being included in a higher ACVIM class (OR=6.383, p-value=0.014). Log-rank analysis showed shorter survival in dogs with low SIC (p=0.020), multivariate Cox analysis revealed that only HF symptoms can affect survival.