Application of a zona pellucida binding assay (ZBA) in the domestic cat benefits from the use of in vitro matured oocytes

<p>Abstract</p> <p>Background</p> <p>Zona pellucida binding assays (ZBAs) have proven useful in determining the fertilising ability of spermatozoa in several species. Most ZBAs use fresh or salt-stored oocytes collected from fresh ovaries but because ovaries are not easy to obtain on a regular basis, chilled and frozen-thawed ovaries have been tested, with varying results. The present study tested the hypothesis that cat spermatozoa, either fresh or frozen-thawed, can bind to homologous zona pellucida of oocytes retrieved from frozen-thawed queen ovaries to a similar extent as they can bind to the zona pellucida of fresh, <it>in vitro </it>matured oocytes.</p> <p>Methods</p> <p>Ovaries were collected from queens after routine ovario-hysterectomy and either stored in NaCl at -20°C until use (treatment animals), or used fresh (controls). Cumulus-oocyte complexes (COCs) were retrieved by ovarian slicing from either source and used directly (immature oocytes from frozen-thawed ovaries; treatment animals) or after <it>in vitro </it>maturation (IVM) (fresh ovaries; controls) for 24 hours in TCM 199, supplemented with 1 IU hCG/mL and 0.5 IU eCG/mL and 0.5% bovine serum albumin (BSA). The oocytes were incubated for 4 hours in 5% CO₂in air at 38°C and 100% humidity in the presence of 5 × 10⁶fresh or frozen-thawed spermatozoa/mL. Representative samples of oocytes were processed for scanning electron microscopy (SEM).</p> <p>Results</p> <p>Both fresh and frozen-thawed spermatozoa bound to the <it>in vitro </it>matured zona pellucida but significantly fewer, or no, spermatozoa bound to frozen-thawed, immature zona pellucida (P < 0.001). Also, more fresh spermatozoa than frozen-thawed spermatozoa bound to the zona pellucida (P < 0.001). The zona pellucida surface differed in morphology (SEM), with <it>in vitro </it>matured oocytes showing a dense surface with few fenestrations in contrast to their frozen-thawed, immature counterparts, where fenestrations were conspicuously larger.</p> <p>Conclusion</p> <p>In conclusion, under the conditions of the present study, immature oocytes recovered from ovaries frozen immersed in NaCl at -20°C are less suitable for use in feline ZBA.</p

Carriage of methicillin-resistant Staphylococcus pseudintermedius in dogs--a longitudinal study

<p>Abstract</p> <p>Background</p> <p>Methicillin-resistant <it>S. pseudintermedius </it>strains (MRSP) are reported with increasing frequency in bacterial cultures from dogs. The objectives of this study were to determine whether MRSP could be found in dogs several months after a clinically apparent infection and whether the length of carriage varied depending on systemic antimicrobial treatment, diagnosis at time of the first positive MRSP culture and the presence of skin disease or wounds. Thirty-one dogs previously diagnosed with a clinical infection were sampled repeatedly for a minimum of eight months or, with the exception of two dogs, until two consecutive negative results were obtained. Five specified locations were sampled, and the results were evaluated to determine future recommendations concerning sample strategies when screening for MRSP carriage. Information was collected from medical records and questionnaires to evaluate factors that may influence length of carriage.</p> <p>Results</p> <p>The overall median length of MRSP carriage was 11 months (48 weeks). The presence of wounds and signs of dermatitis did not influence length of carriage. Systemic treatment for three weeks or longer with antimicrobial agents to which the bacterium was resistant was associated with prolonged carriage compared to dogs treated for a shorter period of time. Three of five dogs treated with an antimicrobial to which their MRSP-isolates were susceptible (tetracycline) were found to still be MRSP-positive when sampled after the end of treatment. Wound samples had the highest positive MRSP yield (81%) for the positive sample sites, compared to less than 70% for each of the other four sample sites. Cultures from the nostrils were less likely to detect MRSP carriage relative to the pharynx, perineum, wounds and the corner of the mouth.</p> <p>Conclusions</p> <p>Dogs can carry MRSP for more than a year after a clinically apparent infection. Systemic antimicrobial treatment of infections with antimicrobial agents to which the MRSP-bacteria are resistant should be avoided when possible in dogs with possible or confirmed MRSP carriage or infection, since it may prolong time of MRSP carriage. Simultaneous sampling of pharynx, perineum, and the corner of the mouth as well as wounds when present is recommended when screening for MRSP. Cultures from nostrils were shown to be less likely to detect MRSP carriage.</p

Differences in metabolic profiles between the Burmese, the Maine coon and the Birman cat-Three breeds with varying risk for diabetes mellitus

Feline diabetes mellitus shares many features with type 2 diabetes in people, regarding clinical presentation, physiology, and pathology. A breed predisposition for type 2 diabetes has been identified, with the Burmese breed at a fivefold increased risk of developing the condition compared to other purebred cats. We aimed to characterize the serum metabolome in cats (n = 63) using nuclear magnetic resonance metabolomics, and to compare the metabolite pattern of Burmese cats with that of two cat breeds of medium or low risk of diabetes, the Maine coon (MCO) and Birman cat, respectively. Serum concentrations of adiponectin, insulin and insulin-like growth factor-1 were also measured (n = 94). Burmese cats had higher insulin and lower adiponectin concentrations than MCO cats. Twenty one metabolites were discriminative between breeds using a multivariate statistical approach and 15 remained significant after adjustment for body weight and body condition score. Burmese cats had higher plasma levels of 2-hydroxybutyrate relative to MCO and Birman cats and increased concentrations of 2-oxoisocaproic acid, and tyrosine, and lower concentrations of dimethylglycine relative to MCO cats. The metabolic profile of MCO cats was characterized by high concentrations of arginine, asparagine, methionine, succinic acid and low levels of acetylcarnitine while Birman cats had the highest creatinine and the lowest taurine plasma levels, compared with MCO and Burmese. The pattern of metabolites in Burmese cats is similar to that in people with insulin resistance. In conclusion, the metabolic profile differed between healthy cats of three breeds. Detection of an abnormal metabolome might identify cats at risk of developing diabetes

Effects of pre-meal drinks with protein and amino acids on glycemic and metabolic responses at a subsequent composite meal.

Whey proteins have insulinogenic properties and the effect appears to originate from a specific postprandial plasma amino acid pattern. The insulinogenic effect can be mimicked by a specific mixture of the five amino acids iso, leu, lys, thr and val

Development and psychometric testing of an instrument to measure the patient’s experience of external radiotherapy : The Radiotherapy Experience Questionnaire (RTEQ)

Background The patient’s perception of external radiotherapy (RT) procedures and equipment is important to evaluate as a complement to endpoints such as treatment outcome and reproducibility. There is a lack of a proper, psychometrically robust instrument to evaluate the patient’s comfort and experience of the external RT procedure. Hence, this study aimed to develop and test an instrument to measure the patient’s experience during external RT. Material and Methods A preliminary 34-item questionnaire was generated from research literature, expert consultations and patient interviews, and it was distributed to patients (n = 825) at 8 RT units in Sweden. The answers were subjected to item analysis and reduction by using exploratory factor analysis. The reliability of the final questionnaire was evaluated using Cronbach’s alpha. Mean scale scores were compared across gender, length of RT and treatment area. Results Most items were highly skewed towards positive responses. Scree plot analyses of the 34-item correlation matrix identified six underlying themes explaining 68% of the total variance. After item reduction, the 6 themes explained 73% of the variance in a 23-item questionnaire. Cronbach’s alpha was satisfactory for all themes (between 0.79 and 0.9). Significant differences between treatment areas were found for two scales: situational unease and situational repose. Conclusion The RT Experience Questionnaire is a tentatively valid and reliable instrument to measure how patients experience the external RT session process and the environment in the treatment room