TOTAL ANOMALOUS PULMONARY VENOUS CONNECTION IN A FOAL

An 8-day-old Arabian-Morgan cross colt underwent cardiac evaluation. The foal was tachycardic, tachypneic, exercise intolerant and had a loud right-sided heart murmur and cyanotic mucous membranes. Total anomalous pulmonary venous connection was diagnosed with echocardiography and confirmed at postmortem examination. Total anomalous pulmonary venous connection is a very rare congenital cardiac abnormality that has not been reported before in the horse.     

Effects of sub-minimum inhibitory concentration antibiotic levels and temperature on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus

The objective of this study was to determine the effects of sub-minimum inhibitory concentrations (sub-MICs) of 2 veterinary antibiotic preparations, chlortetracycline (CTC) and chlortetracycline-sulfamethazine (CTC + SMZ), on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus at normal and febrile body temperatures. Sub-minimum inhibitory concentrations of both antibiotics reduced the growth rates of M. haemolytica and H. somnus. Growth of both species was not inhibited when grown at 41 degrees C compared to 37 degrees C. There was no detectable consistent effect of antibiotic or temperature on outer membrane protein expression for either species. Our study indicates that sub-MIC levels of CTC and CTC + SMZ markedly impair growth of clinical M. haemolytica and H. somnus isolates, potentially allowing more effective host clearance during infection.     

Mixed models applied to the study of variation of grower-finisher mortality and culling rates of a large swine production system

Large scale production systems for swine are frequently organized in a hierarchical structure. Consequently, important production parameters, such as mortality and culling, can be analyzed at different levels. The major aims of this study were to assess variance components (VC) of mortality and culling rates attributed to sites and to barns within a site, and subsequently to investigate the impact of average entry weight, days on feed (length of the production turn), and season on the magnitude of the VC. Then, data from a large farm with 3 sites were collected during 5 y. In total, 1720040 pigs distributed in 1502 all-in/all-out grower-finisher groups were included. Linear mixed models were fitted for mortality and culling rates. The barn was modeled as the residual component (barn-to-barn variations) with production turn and site nested within production turn as random intercept variance components. Barn-to-barn pig group variation was the largest VC for mortality (63.08%), when no predictors were included in the models. Predictors, such as pigs placed on quarters 2 and 3, low average entry weight, and shorter production turn length, were associated together with higher mortality. The explained proportion of variance due to these predictors was about 12.05% and the VC for barn, site, and production turn were 67.6%, 17.6%, and 14.8%, respectively. Barn-to-barn variation was also the largest VC for culling rate (46.2%), but the same predictor mentioned above explained only about 1.4% of the variation. The VC for barn, site, and production turn were 46.8%, 21.3%, and 31.8%, respectively. Since the variability among barns far exceeded the variability among sites, the barn should be used as experimental unit in studies with grower-finisher mortality, culling rate, or both, as outcome variables.     

Apoptosis in the Antral Follicles of Swamp Buffalo and Cattle Ovary: TUNEL and Caspase-3 Histochemistry

The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Malignant catarrhal fever-like disease in sheep after intranasal inoculation with ovine herpesvirus-2.

A malignant catarrhal fever (MCF)-like disease was induced experimentally in 3 sheep after aerosol inoculation with ovine herpesvirus-2 (OvHV-2). Each of 3 OvHV-2-negative sheep was nebulized with 2 ml of nasal secretions containing approximately 3.07 X 10(9) OvHV-2 DNA copies from a sheep experiencing an intensive viral-shedding episode. Ovine herpesvirus-2 DNA became detectable by polymerase chain reaction in the peripheral blood leukocytes of all 3 sheep within 3 days, and all 3 seroconverted between 6 and 8 days postinfection (PI). The sheep developed clinical signs, with copious mucopurulent nasal discharge and fever around 14 days PI. One of the 3 clinically affected sheep was euthanized at 18 days PI. Major lesions at necropsy were multifocal linear erosions and ulcers in mucosa of the cheeks, tongue, pharynx, and proximal esophagus and mild disseminated pneumonia. Microscopically, there was extensive moderate superficial histiocytic-lymphocytic rhinitis with epithelial dissociation and degeneration. Moderate multifocal histiocytic bronchointerstitial pneumonia was associated with loss of terminal bronchiolar epithelium. Lymphocytic vasculitis was present only in the lung. The remaining 2 sheep recovered clinically, approximately 25 days PI. The study revealed that clinical signs and lesions resembling MCF can develop when uninfected sheep are exposed to a high dose of aerosolized OvHV-2.     

Detection of Echinococcus multilocularis infection in a colony of cynomolgus monkeys (Macaca fascicularis) using serology and ultrasonography.

Five animals in a colony of cynomolgus monkeys (Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.     

Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Predicting the spatial distribution of ground flora on large domains using a hierarchical Bayesian model

Accomodation of important sources of uncertainty in ecological models is essential to realistically predicting ecological processes. The purpose of this project is to develop a robust methodology for modeling natural processes on a landscape while accounting for the variability in a process by utilizing environmental and spatial random effects. A hierarchical Bayesian framework has allowed the simultaneous integration of these effects. This framework naturally assumes variables to be random and the posterior distribution of the model provides probabilistic information about the process. Two species in the genus Desmodium were used as examples to illustrate the utility of the model in Southeast Missouri, USA. In addition, two validation techniques were applied to evaluate the qualitative and quantitative characteristics of the predictions.This revised version was published online in May 2005 with corrections to the Cover Date.     

Assessing the potential impacts of alternative landscape designs on amphibian population dynamics

An individual-based, spatially explicit population model was used to predict the consequences of future land-use alternatives for populations of four amphibian species in two central Iowa (midwest USA) agricultural watersheds. The model included both breeding and upland habitat and incorporated effects of climatic variation and demographic stochasticity. Data requirements of the model include life history characteristics, dispersal behavior, habitat affinities, as well as land use and landcover in geographic information systems databases. Future scenarios were ranked according to change in breeder abundance, saturation, and distribution, compared to baseline conditions. Sensitivity of simulation results to changes in model parameters was also examined. Simulated results suggest that while all four species modeled are likely to persist under present and future scenario conditions, two may be more at risk from future landscape change. Although the study species are all widespread generalists regarded as having a low conservation priority, they depend on wetlands and ponds, increasingly endangered habitats in agricultural landscapes. Broader conservation strategies in the region would ensure that these currently common organisms do not become the endangered species of the future.This revised version was published online in May 2005 with corrections to the Cover Date.