Haematological and biochemical investigations of diseased koalas (Phascolarctos cinereus)

Haematological and biochemical investigations were performed on 14 koalas with uncomplicated cystitis, 8 with complicated cystitis, 8 with conjunctivitis, 8 with lymphosarcoma, and 14 with miscellaneous diseases. Changes were limited and inconsistent in individual koalas with uncomplicated cystitis and conjunctivitis. In contrast, individual koalas with complicated cystitis were more likely to have anaemia, leukocytosis due to neutrophilia, hypoproteinaemia due to hypoalbuminaemia, and azotaemia due to elevated urea concentration. Although these changes were non-specific they did allow assessment of prognosis for survival and response to treatment. Koalas with lymphosarcoma were invariably anaemic, leukaemic, azotaemic and hypoalbuminaemic. Elevated enzymes (aspartate transaminase [AST]. lactate dehydrogenase [LD] and gamma glutamyl transferase [GGT]) were more common in koalas with lymphosarcoma. Koalas affected by miscellaneous conditions showed variable changes but once again anaemia, leukocytosis, azotaemia, elevated AST and LD, and hypoalbuminaemia were not uncommon. On the basis of these findings a minimal profile is suggested for the investigation of sick koalas and would include haematocrit, total and differential leukocyte counts, urea, total protein and albumin concentrations and AST, GGT and LD activities.     

Randomly primed,strand-switching,MinION-based sequencing for the detection and characterization of cultured RNA viruses

RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.     

Analysis of cerebrospinal fluid from dogs and cats after 24 and 48 hours of storage

OBJECTIVE: To compare differential cell counts and cell characteristics of CSF samples analyzed immediately or after storage for 24 and 48 hours at 4 C with and without the addition of autologous serum. DESIGN: Prospective study. ANIMALS: 36 dogs and 6 cats. PROCEDURE: CSF samples were collected from the cerebellomedullary cistern and divided into 250-microliter aliquots. Slides of CSF samples were prepared by use of cytocentrifugation immediately and after 24 and 48 hours of storage with addition of autologous serum (final concentrations, 11 and 29%). Differential cell counts and number of unrecognizable cells were compared among preparations. RESULTS: Significant differences in the differential cell counts were not detected among samples analyzed before or after storage. Although the number of unrecognizable cells increased with storage time, this did not result in a significant effect on cell distribution or diagnosis. Cells in CSF samples stored with 11% serum more closely resembled cells in fresh samples than did cells in samples stored with 29% serum. CONCLUSIONS AND CLINICAL RELEVANCE: CSF samples collected at veterinary clinics remote from a diagnostic laboratory or during nonoperational hours may be preserved through the addition of autologous serum. Evaluation of such samples is likely to result in an accurate diagnosis for at least 48 hours after collection.     

Simulation study of the competitive ability of erect, semi-erect and prostrate cowpea (Vigna unguiculata) genotypes

Ecophysiological simulation models provide a quantitative method to predict the effects of management practices, plant characteristics and environmental factors on crop and weed growth and competition. The INTERCOM interplant competition model was parameterised, calibrated by monoculture data for three cowpea (Vigna unguiculata) genotypes that differed in growth habit, common sunflower (Helianthus annuus) and common purslane (Portulaca oleracea), and used to simulate competition of cowpea cover crops with sunflower or purslane. The simulation results were compared with observations from field competition experiments in 2003 and 2004. INTERCOM more accurately simulated actual field data for the competition of cowpea genotypes and sunflower than companion field experiments for the competition of cowpea and purslane. The validated simulation model of cowpea and sunflower at two densities was used to study the effects of cowpea growth habit on final biomass production of cowpea and sunflower. The model suggested that erect growth habit was more competitive than semi‐erect and prostrate growth habit, when cowpea genotypes were grown with sunflower. Cowpea leaf area distribution was important to higher cowpea biomass production, while cowpea height growth was important to reduce sunflower biomass. Our simulation approach is suggested as a method for crop breeders to gauge the likely success of selection for competitive crops before undertaking expensive long‐term breeding experiments.     

Gonadotropin‐induced Puberty Does Not Impair Reproductive Performance of Gilts over Three Parities

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.     

Influence of a probiotic adjunct culture of Enterococcus faecium on the quality of cheddar cheese

Cheddar cheese has previously been shown to be an effective vehicle for delivery of viable cells of a probiotic Enterococcus faecium strain to the gastrointestinal tract. The particular strain, E. faecium PR88, has proven efficacy in the treatment of irritable bowel syndrome, and in this study it was evaluated for suitability as a starter adjunct for Cheddar cheese manufacture. When added to cheesemilk at an inoculum of 2 x 10(7) cfu/mL, the enterococcal adjunct maintained viability in Cheddar cheese at levels of up to 3 x 10(8) cfu/g during 9 months of ripening. Increased proteolysis and higher levels of some odor-active volatile compounds were observed in Cheddar cheeses containing the PR88 adjunct compared with the control throughout the ripening period. In addition, the enterococcal adjunct strain did not affect cheese composition. Although sensory evaluation showed no significant difference in flavor/aroma and body/texture scores between control and experimental cheeses, repeated comments by the commercial grader consistently described the cheeses containing PR88 as 'more advanced than the control' and as having 'better flavor'. These findings indicate that the presence of the PR88 adjunct strain in Cheddar cheese at levels of >/=10(8) cfu/g may positively influence Cheddar flavor.