Comparison of Whole Ovary Cryotreatments for Fertility Preservation

The goal of this study was to compare a traditional slow‐freeze method (TF) with an open unidirectional slow freeze cooling system (UF) for whole ovary cryopreservation. Therefore, whole pig ovaries were randomly assigned to (A) fresh control, (B) traditional slow freeze (TF) or (C) unidirectional slow freeze (UF). Ovaries were perfused with 10% DMSO in Krebs‐Ringer. For TF, whole ovaries were placed in specimen jars containing 10% DMSO and placed into a specialized container for freezing filled with propan‐2‐ol. For UF, whole ovaries were placed within a specially designed container containing 10% DMSO and transferred to a specialized freezing machine (CTE 920). Histological evaluation demonstrated intact morphology of follicles in all groups; however, an overall decrease of follicle numbers in TF (46%) and UF (50%) compared to fresh control. Live/dead assay indicated significantly lower populations of live cells in both TF (60%) and UF (58%) compared to fresh tissue (74%). TUNEL assay confirmed a difference in percentage of apoptotic follicles between fresh and TF, but there was no significant difference between fresh and UF. To improve the structural and functional integrity of whole ovaries, further investigation, especially into directional freezing, is needed. Whole ovary cryopreservation could provide opportunities for women facing fertility loss due to chemo‐ or radiotherapy treatment.     

Elevated temperature affects growth and hemato‐biochemical parameters,inducing morphological abnormalities of erythrocytes in Nile tilapia Oreochromis niloticus

Temperature is considered as an important environmental factor, and the increasing water temperature resulting from global warming is a great concern. The present study was conducted to examine the effects of elevated water temperature on growth, hemato‐biochemical parameters in Nile tilapia, Oreochromis niloticus acclimatized to three temperatures (31°C, 34°C and 37°C) for 60 days. Additionally, erythrocytic cellular abnormalities (ECA) and erythrocytic nuclear abnormalities (ENA) tests were assayed using peripheral erythrocytes after exposure to the three temperatures. Fish were sacrificed on days 7, 15, 30 and 60 of exposure. Growth performances viz., weight gain, % weight gain and specific growth rate (SGR) showed decreasing tendency at 34°C but significantly declined at 37°C compared to 31°C. The abundance of haemoglobin (Hb) and red blood cells (RBCs) significantly decreased in response to temperature increases, while white blood cells (WBCs) displayed the opposite response. At days 7 and 15, blood glucose levels significantly increased in response to the temperature increase, while at days 30 and 60 glucose declined. Frequencies of ECA and ENA were significantly enhanced at the highest temperature throughout the experimental period. Dissolved oxygen decreased and free CO₂ increased significantly with increasing temperature throughout the study period. The present study revealed that temperatures higher than 34°C may be hazardous to O. niloticus.     

Immune response and reactions to various dose regimens for raising hyperimmune antisera in sheep

To determine the optimum procedure for raising hyperimmune sera to tetanus toxin, three adjuvants, four antigen preparations and two routes of administration in various combinations were investigated in sheep. Oil-in-water adjuvants alone or in combination with aluminum gels were superior to aluminium gels on their own. This disadvantage of aluminium gels was partially but not completely abrogated when the frequency of doses was increased to three per week. Intensity of local reaction was strongly correlated with immune response; the more immunogenic a dose, the more reactive. Reactivity of oily adjuvants could be lessened by use of a more suitable route of administration, thus oily adjuvants appeared suitable for use when administered by the intraperitoneal route even though moderate to severe reactions resulted from subcutaneous injections. Of other variables investigated, toxin did not confer any advantage over toxoid as an immunogen, purified toxoid was a significantly better immunogen than unpurified toxoid and two large bleeds (30% of total blood volume each) every six weeks rather than 20 ml test bleeds did not affect the titre of the hyperimmune serum produced.     

Malignant lymphoma in a Sinclair miniature pig

Malignant lymphoma was diagnosed in a 3-year-old, male Sinclair(S-1) miniature pig with acute anorexia, depression, fever, and markedly enlarged inguinal lymph nodes. Results of an initial hemogram indicated a leukocyte count of 121,489 cells/mm3. Most of the leukocytes were mononuclear cells of various sizes, nuclear chromatin pattern, number of nucleoli, amount of cytoplasm, and amount of staining. Cytochemical staining and flow cytometric evaluation of the leukocytes indicated a large number of hypodiploid lymphoblasts in the peripheral blood. Gross necropsy findings included enlargement of all lymph nodes, a pale liver, and multifocal pale areas scattered throughout the kidneys. Microscopic examination indicated massive infiltration of abnormal lymphoid cells into most major organs and complete loss of normal morphologic features of all lymph nodes.     

Growth Hormone-producing Pituitary Adenoma, Elevated Serum Somatomedin C Concentration and Diabetes Mellitus in a Cat

A pituitary acidophil adenoma in a domestic shorthaired cat with diabetes mellitus and elevated serum somatomedin C level is described. Growth hormone production by the neoplasm was confirmed by an appropriate reaction using an immunoperoxidase technic.     

Resident forum abstractsEVALUATION OF THROMBOELASTOGRAPHY (TEG) IN NORMAL CATS

Objective: To establish normal parameters of thromboelastography (TEG) in healthy adult cats. Background: Thromboelastography (TEG) is an in vitro test of coagulation that has been shown to be useful in humans, dogs and select species to identify and quantify alterations of hemostasis (e.g., hypercoagulable and hypocoagulable states). It has also been demonstrated to be useful in monitoring effects of anticoagulant therapies. This test has not been evaluated in cats. Methods: Blood was collected from 25 clinically normal cats by venipuncture using a 21 gauge×3 1/2 inch butterfly catheter and syringe for medial saphenous or jugular venipuncture. A single 1.8 mL sample in 3.8% Sodium Citrate (9:1) was collected from each cat. Recalcified whole blood was analyzed 30 minutes following collection with the TEG® 5000 analyzer (Haemoscope, Niles, IL). Analysis temperature was 37.6°C. TEG parameters recorded included: R‐value (represents initial fibrin formation), K (time from R to standard fixed measure of clot firmness which represents contributions of platelets and fibrinogen), maximum amplitude (MA; represents absolute clot strength), and alpha angle (α; the slope of TEG tracing which represents rate of clot formation). The coagulation index (CI) was derived from the formula generated for humans to provide an overall assessment of whether the sample was hyper‐ or hypocoagulable. Results: Values for the 25 normal cat samples are reported as mean ±2 standard deviations. R=2.97; 1.23–4.72; K=1.54, 0.38–2.71; α=70.70, 57.76–83.65; MA=58.50, 45.26–71.74 and CI=2.27, 0.07–4.46. Compared to historical information obtained on normal dogs, cats have significantly shorter R and K and larger α, MA and CI. Conclusions: TEG does have reproducible performance when used to evaluate coagulation status in normal cats. Compared to dogs, normal cats favor a hypercoagulable state. Species‐specific normal values are necessary for interpretation of TEG results. This test bears potential value for use in future experimental and clinical work to investigate hemostasis in cats receiving anticoagulant therapies or in cats suffering from diseases such as cardiomyopathy which are thought to be associated with altered coagulation status.     

Isolation and identification of AIDA-I receptors in porcine intestinal mucus

Porcine AIDA-I positive Escherichia coli causes diarrhea in neonatal piglets and AIDA-I adhesin is an important virulence factor involved in intestinal colonization with biofilm formation. This biofilm consists of AIDA-I(+)E. coli bacteria stratified within mucus layers covering the intestinal mucosa. Based on the intimate interaction between AIDA-I(+)E. coli and mucus within the intestinal biofilm, we hypothesized that porcine intestinal mucus contains receptor(s) for AIDA-I adhesin. Since porcine AIDA-I receptors have not been identified, we employed affinity chromatography and in vitro adhesion assays to investigate AIDA-I binding proteins in porcine intestinal mucus that might serve as receptors for attachment of AIDA-I positive E. coli. We demonstrated that porcine mucus contains 65 and 120kDa proteins (p65 and p120) that bind with high affinity to purified AIDA-I adhesin and that AIDA-I positive E. coli binds to these proteins with higher affinity than do AIDA-I negative mutant. The identity of p65 was not determined based on LC-MS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.     

Physiological condition and migratory experience affect fitness‐related outcomes in adult female sockeye salmon

Relating fish physiology, behaviour and experience to fitness‐related outcomes at the individual scale is ecologically significant, but presents difficulties for free‐ranging fishes in natural systems. Physiological state (e.g. level of stress or maturity) and experience (e.g. habitat use or exposure to stressors) may alter the probability of survival or reproduction. This study examined the relative influence of physiology and migratory experience on survival, migration duration, reproductive longevity, and egg retention in adult female sockeye salmon (Oncorhynchus nerka) from a Fraser River population. One hundred and thirty‐five females were plasma sampled and tagged with radio transmitters and archival temperature loggers. Fish were tracked 55 km through two natal lakes to spawning grounds, following passage of a hydroelectric dam. For 39 females, we assessed the proportion of time within an optimal temperature (T_optAS) window (13.4–19.5°C), which provides ≥90% of maximum aerobic scope. Females with lower plasma glucose concentrations were more likely to reach spawning grounds. Early migrants spent longer in natal lakes. More time in the T_optAS window was associated with greater reproductive longevity and lower probability of egg retention. Later arriving females had reduced longevity on spawning grounds, as did females that retained eggs. Exposure to higher dam discharge was associated with reduced reproductive longevity and greater probability of egg retention, but not lower survival, indicating a delayed effect of dam passage. Our results underscore the complexity of factors governing fitness‐related outcomes for salmonids, particularly the importance of female experience in the days and weeks prior to spawning.