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.     

Dam Mycobacterium avium subspecies paratuberculosis (MAP) infection status does not predetermine calves for future shedding when raised in a contaminated environment: a cohort study

Uptake of Mycobacterium avium subsp. paratuberculosis (MAP) by calves in the first days of life from colostrum, milk and faeces is regarded an important moment of transmission. The objective of this study was to quantify the association between the MAP status of dams as determined by the presence of MAP DNA and antibody in colostrum and that of DNA in faeces and the environment with subsequent MAP shedding of their daughters. A cohort of 117 dam-daughter pairs giving birth/being born on eight commercial dairy farms with endemic paratuberculosis was followed where colostrum, faecal and environmental samples (dust) were analysed for the presence of MAP using an IS900 real-time PCR. Antibodies in colostrum were measured by ELISA. Analysis of dust samples showed that on all farms environmental MAP exposure occurred continuously. In significantly more colostrum samples (48%) MAP DNA was detected compared to faecal samples (37%). MAP specific antibodies were present in 34% of the colostrum samples. In total MAP DNA was present in faecal samples of 41% of the daughters at least once during the sampling period. The association between faecal shedding in the offspring and the dam MAP status defined by MAP PCR on colostrum, MAP PCR on faeces or ELISA on colostrum was determined by an exact cox regression analysis for discrete data. The model indicated that the hazard for faecal shedding in daughters born to MAP positive dams was not significantly different compared to daughters born to MAP negative dams. When born to a dam with DNA positive faeces the HR was 1.05 (CI 0.6; 1.8) and with DNA positive colostrum the HR was 1.17 (CI 0.6; 2.3). When dam status was defined by a combination of both PCR outcomes (faeces and colostrum) and the ELISA outcome the HR was 1.26 (CI 0.9; 1.9). Therefore, this study indicates that neither the presence of MAP DNA in colostrum, MAP DNA in faeces nor the presence of MAP antibodies in colostrum of the dam significantly influences the hazard of MAP shedding in their subsequent daughters up to the age of two years when raised in a contaminated environment.     

Biogeochemical Controls and Feedbacks on Ocean Primary Production

Changes in oceanic primary production, linked to changes in the network of global biogeochemical cycles, have profoundly influenced the geochemistry of Earth for over 3 billion years. In the contemporary ocean, photosynthetic carbon fixation by marine phytoplankton leads to formation of approximately 45 gigatons of organic carbon per annum, of which 16 gigatons are exported to the ocean interior. Changes in the magnitude of total and export production can strongly influence atmospheric CO2 levels (and hence climate) on geological time scales, as well as set upper bounds for sustainable fisheries harvest. The two fluxes are critically dependent on geophysical processes that determine mixed-layer depth, nutrient fluxes to and within the ocean, and food-web structure. Because the average turnover time of phytoplankton carbon in the ocean is on the order of a week or less, total and export production are extremely sensitive to external forcing and consequently are seldom in steady state. Elucidating the biogeochemical controls and feedbacks on primary production is essential to understanding how oceanic biota responded to and affected natural climatic variability in the geological past, and will respond to anthropogenically influenced changes in coming decades. One of the most crucial feedbacks results from changes in radiative forcing on the hydrological cycle, which influences the aeolian iron flux and, in turn, affects nitrogen fixation and primary production in the oceans.     

Failure of an in vitro lymphoproliferative assay specific for bovine herpes virus type 1 to detect immunised or latently infected animals

The in vitro lymphoproliferative assay specific for bovine herpes virus type 1 (BHV1) was tested for its ability to predict whether an animal was protected against challenge with virulent BHV1 and for its ability to identify animals latently infected with the virus. Three animals that had been in contact with a field strain of the virus, three that had been vaccinated with a modified live-virus vaccine seven weeks previously, six that had been vaccinated in the same way five months previously, and seven control animals that had had no previous contact with the virus were challenged with virulent BHV1. The 12 animals that had had previous contact with BHV1 all resisted the challenge well or fairly well, but six of them did not react positively in the in vitro lymphoproliferative assay. It was concluded that the assay did not give consistent evidence of the immune status of the animals. Four animals that had had previous contact with a field strain of BHV1 were treated with dexamethasone; they excreted BHV1 irrespective of whether they showed a positive response in the in vitro lymphoproliferative assay.     

Efficacy of ivermectin jetting fluid for the curative treatment of blowfly strikes in sheep

SUMMARY A jetting fluid containing 7.5% w/v ivermectin, which was diluted to 0.03 mg/mL in water and applied using a standard hand jetting technique, was assessed for its efficacy in treating blowfly strikes on Merino sheep. Strikes were induced on sheep held under insectary conditions using laboratory-reared Lucilia cuprina. All struck sheep jetted at 28,48 or 72 hours after exposure to flies were successfully treated. All 1st, 2nd and early 3rd instar larvae were killed within 24 hours of treatment.     

Land-use changes by large-scale plantations and their effects on soil organic carbon,micronutrients and bulk density: empirical evidence from Ethiopia

This article examines land-use changes by large-scale plantations in Ethiopia and evaluates the impacts thereof on soil organic carbon, micronutrients and bulk density. Remote sensing analysis and field research activities were undertaken at four large-scale plantation projects in Benshanguel Gumuz, Gambella, and Oromia regional states. Results show that the projects largely involved the conversion of both closed and open to closed forests and grasslands, which in turn reduced soil carbon stock and micronutrient levels and increased soil compaction. We argue that unless appropriate soil management activities and impact mitigation strategies are adopted by plantation proponents, these land-use changes will pose a serious threat to the long-term economic viability and sustainability of plantation agriculture in Ethiopia. This could undermine long-term ecosystem health and national food security.     

Neutrophil migration in the lung,general and bovine-specific aspects

Lung inflammation is often associated with sustained neutrophil migration into the lung tissue, causing undesired side effects, i.e. substantial damage of lung tissue and fibrin deposition, which hamper complete recovery. The need for additional anti-inflammatory treatment strategies focused attention on the function of cellular adhesion molecules (CAMs) on the leukocyte membrane, which guide migration of leukocytes across the endothelium to the site of inflammation. Recent data indicate that neutrophil migration in the lung is mediated by unique pathways, involving different CAM as compared to other organs. These pulmonary characteristics of neutrophil migration enable specific targeting of CAM for anti-inflammatory treatment of pneumonia. In addition, the recent interest in intracellular signaling revealed that most CAM not only function in adhesion, but also play an important role in initiation of intracellular signal transduction and vice versa may be modulated by intracellular signaling molecules (ISMs). In this review, the mechanisms of neutrophil migration in the lung and the role of ISM with respect to CAM function are described in the context of potential anti-inflammatory intervention strategies.     

High volume continuous venovenous haemofiltration (HV-CVVH) in an equine endotoxaemic shock model

Equine acute abdominal disease is often associated with shock. Important aspects in the onset of this complication include hypovolaemia, the translocation of endotoxins from the gut and the subsequent activation of the cytokine network. The clinical efficacy of high volume continuous venovenous haemofiltration (HV-CVVH) and the clearance of cytokines were therefore investigated in an equine endotoxaemic model. Ten male Shetland ponies received a slow infusion of LPS (2 microg/kg bwt) under general anaesthesia. The treatment group (n = 5) received HV-CVVH (2 ml/kg bwt/min) using a 75 kD polymethylmethacrylate (PMMA filter). Haemodynamic, blood variable and cytokine (TNF, IL-1 and IL-6) measurements were performed every 30 min for a 6 h period. The ponies showed a typical reaction in mean pulmonary arterial pressure, blood chemical and haematological markers after LPS challenge. No significant differences were found between the treatment group and the control group. Only a slight increase in cardiac index and no marked decrease in mean arterial pressure were seen. A clear cytokine response was found in all ponies, though substantially different in magnitude between individuals. The clearance of cytokines from the blood increased in time, but did not lead to significant decrease in serum levels. In this study, HV-CVVH with a PMMA filter did not prove to have a significant beneficial effect on the course of experimental endotoxaemia in horses. However, in a more severe model, better efficacy might be obtained. Testing additional filters might lead to a more suitable therapy for horses.     

Lewis rats are not susceptible to oral infection with Mycobacterium avium subsp. paratuberculosis

Pathogenesis studies of Mycobacterium avium subsp. paratuberculosis infection in ruminants are hampered by the long incubation time of the disease. A laboratory animal model with a shorter incubation time would facilitate research in this field. Although small rodents are usually considered to be resistant to M.a. paratuberculosis infection, several susceptible murine strains have been found. To our knowledge, there are no detailed reports with regard to susceptibility in rats. The Lewis rat is a valuable model for inflammatory bowel disease studies as well as autoimmune diseases involving mycobacteria as inducing agents. In this study Lewis rats were used to investigate their potential as a small laboratory animal model for paratuberculosis.In total 28 female Lewis rats were orally inoculated with M.a. paratuberculosis. The rats were first inoculated at 3 weeks of age, and 12 more inoculations followed in increasing intervals during the 3 months to follow. Eight control rats received a sham inoculation. Over 9 months, two rats from each group were sacrificed at regular intervals and immunological and histopathological examinations were performed on the gastrointestinal tract, the liver and the spleen.None of the rats developed lesions which were indicative of mycobacterial infection as determined by histology with HE and Ziehl-Neelsen staining. The bacteria could not be recultured from samples taken from the gut, the liver or the spleen. The immunological tests however, showed that bacteria had entered via the intestinal tract. From this study it appears that Lewis rats are resistant to oral inoculation with M. a. paratuberculosis, and not suitable as a model to study the immunopathogenesis of paratuberculosis as it occurs in ruminants.