Waking experience affects sleep need in Drosophila

Sleep is a vital, evolutionarily conserved phenomenon, whose function is unclear. Although mounting evidence supports a role for sleep in the consolidation of memories, until now, a molecular connection between sleep, plasticity, and memory formation has been difficult to demonstrate. We establish Drosophila as a model to investigate this relation and demonstrate that the intensity and/or complexity of prior social experience stably modifies sleep need and architecture. Furthermore, this experience-dependent plasticity in sleep need is subserved by the dopaminergic and adenosine 3',5'-monophosphate signaling pathways and a particular subset of 17 long-term memory genes.     

Clostridium piliforme infection in two farm-raised white-tailed deer fawns (Odocoileus virginianus) and association with copper toxicosis

Necropsy of 2 white-tailed deer fawns who died acutely revealed diarrhea and melena in case No. 1 and no gross changes in case No. 2. Histologically, the livers of both deer displayed multifocal coagulative necrosis, with infiltrations of neutrophils, macrophages, and lymphocytes. By Warthin-Starry staining, bundles of filamentous bacteria were identified within hepatocytes at the periphery of the necrotic foci in case No. 1. There was multifocal myocardiocyte necrosis in case No. 1 and multifocal lymphoid necrosis of the Peyer's patches in case No. 2. Clostridium piliforme 16S ribosomal ribonucleic acid gene was detected in both livers by polymerase chain reaction (PCR) with C. piliforme-specific primers. The liver copper levels in both cases were normal to slightly elevated. The kidney copper level in case No. 2 was elevated. This represents the first published cases of Tyzzer's disease in deer, a novel use of PCR for the diagnosis of C. piliforme infection, and a possible association between copper toxicosis and Tyzzer's disease.     

Novel diabetes mellitus treatment: mature canine insulin production by canine striated muscle through gene therapy

Muscle-targeted gene therapy using insulin genes has the potential to provide an inexpensive, low maintenance alternative or adjunctive treatment method for canine diabetes mellitus. A canine skeletal muscle cell line was established through primary culture, as well as through transdifferentiation of canine fibroblasts after infection with a myo-differentiation gene containing adenovirus vector. A novel mutant furin-cleavable canine preproinsulin gene insert (cppI4) was designed and created through de novo gene synthesis. Various cell lines, including the generated canine muscle cell line, were transfected with nonviral plasmids containing cppI4. Insulin and desmin immunostaining were used to prove insulin production by muscle cells and specific canine insulin ELISA to prove mature insulin secretion into the medium. The canine myoblast cultures proved positive on desmin immunostaining. All cells tolerated transfection with cppI4-containing plasmid, and double immunostaining for insulin and desmin proved present in the canine cells. Canine insulin ELISA assessment of medium of cppI4-transfected murine myoblasts and canine myoblast and fibroblast mixture proved presence of mature fully processed canine insulin, 24 and 48 h after transfection. The present study provides proof of principle that canine muscle cells can be induced to produce and secrete canine insulin on transfection with nonviral plasmid DNA containing a novel mutant canine preproinsulin gene that produces furin-cleavable canine preproinsulin. This technology could be developed to provide an alternative canine diabetes mellitus treatment option or to provide a constant source for background insulin, as well as C-peptide, alongside current treatment options.     

Anomalous Pregnancies during Late Embryonic/Early Foetal Period in High Producing Dairy Cows

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004–2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35–41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.     

Serum Profiles of Pregnancy-Associated Glycoprotein, Oestrone Sulphate and Progesterone During Gestation and Some Factors Influencing the Profiles in Ethiopian Borana and Crossbred Cattle

This study presents serum concentrations of pregnancy‐associated glycoprotein (PAG), oestrone sulphate (E1‐S) and progesterone (P4), and the effects of some dam and foetus‐related factors on these profiles during gestation in Borana and crossbred cattle. The PAG concentrations at 4th week post‐conception ranged from 1.5–5.5 and 2.1–4.7 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The mean PAG concentrations increased progressively from 4th to 33rd week of gestation (from 3.3–173 ng/ml for Borana and 4.2–240 ng/ml for crossbred cattle) and reached peak around calving. Breed, parity status, dam body weight, foetal sex and foetal birth weight significantly influenced the PAG concentrations. After delivery, the PAG concentrations declined steadily to 5.7 ng/ml in Borana (n = 7) and 3.9 ng/ml in crossbred (n = 6) cattle 10 weeks post‐partum. The serum E1‐S concentrations at 17th week of pregnancy ranged from 0.3–2.6 and 0.9–5.7 ng/ml in Borana (n = 8) and crossbred (n = 9) cattle, respectively. The mean E1‐S concentrations increased progressively from 17th to 33rd week of gestation (from 1.1–4.6 ng/ml for Borana and 2.7–10.8 ng/ml for crossbred). Breed, parity status, dam body weight and foetal sex significantly influenced E1‐S concentrations. The P4 concentrations at 4th week of pregnancy ranged from 3.2–5.1 and 1.7–8.9 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The P4 level remained elevated throughout pregnancy. This study indicated that the serum PAG and P4 concentrations at 4th and E1‐S approximately 17th week of pregnancy were above the cut‐off value for pregnancy test and the hormonal profiles observed were comparable to the previous reports. Furthermore, the PAG and E1‐S profiles were considerably influenced by factors such as breed, weight and parity status of the dam, and foetal sex and foetal birth weight (only PAG).     

Pre-breeding Ovaro–Uterine Ultrasonography and its Relationship with First Service Pregnancy Rate in Seasonal-Calving Dairy Herds

The objectives of the study were to characterize an ultrasound reproductive tract scoring (URTS) system to assess suitability for breeding in dairy cows, to describe the prevalence of these scores in commercial dairy herds and to examine their relationship with subsequent fertility. Ultrasound examinations (7797) were performed on 5751 Holstein–Friesian cows prior to breeding in 62 seasonally calving herds over 2 years. Data recorded from images of both ovaries and the uterus were combined into a six point scoring system and the prevalence of cows with cystic ovarian follicles and uterine abscesses and adhesions was recorded separately. The prevalence of ovulatory cows which had completed uterine involution (score 1), or had mild (2) or moderate endometritis (3) was 62.5%, 21.7% and 2.2%, respectively. The prevalence of anovulatory cows with moderate endometritis (4), ovulatory cows with pyometra (5) and anovulatory cows which had completed uterine involution (6) was 3.3%, 2.2% and 8.1%, respectively. The interval between calving and examination differed between cows with each of the scores 1, 2, 5 and 6 (61, 46, 53 and 50 days, respectively, p < 0.05) but not between cows with scores 3 and 4 (37 and 35 days, respectively). Ovulatory cows which had completed uterine involution (score 1) had a higher likelihood of pregnancy to first service than ovulatory or anovulatory cows which had not completed uterine involution (p ≤ 0.10). The prevalence of cows with cystic ovarian follicles or abscesses or adhesions of the reproductive tract was 3.9% and 1.2%, respectively. In conclusion, 29% and 11% of cows in seasonally calving and breeding dairy herds had not completed uterine involution or were anovulatory prior to the mating start date, respectively. Both conditions, detected using a URTS system, significantly reduced first service pregnancy rate in these pasture-based dairy herds.     

Use of wound soaker catheters for the administration of local anesthetic for post-operative analgesia: 56 cases

Objective  To describe the administration of local anesthetic through wound soaker catheters for post-operative veterinary patients and to characterize complications.
Study design  Retrospective study of hospital records.
Animals  Records of patients in which a wound soaker catheter was placed post-operatively between November 1, 2004 and July 1, 2006 at a veterinary teaching hospital. Records in which a limb amputation was performed between January 1, 2002 and August 1, 2007 and in which a wound soaker catheter was not placed were reviewed for historic control.
Results  A total of 56 cases were identified in which a wound soaker catheter was placed post-operatively including 52 dogs, 2 cats, and 2 goats. Twenty canine cases were identified in which limb amputation was performed and no wound soaker catheter was placed. The majority of surgical procedures for which a wound soaker catheter was placed included thoracic limb amputation (46.4%) and pelvic limb amputation (35.7%). Wound soaker catheters remained in place for an average of 1.6 ± 0.5 days. Feline and caprine patients received intermittent bupivacaine boluses every 6 hours. Canine patients received continuous lidocaine infusions. Complications included disconnection of the catheter from the infusion (7.7%), one seroma, and one suspected lidocaine neurotoxicity. Incisional infections were noted in 3/56 (5.3%) limb amputations with wound soaker catheters placed which was not higher than the incisional infection rate found in the historic control cases 3/20 (15%).
Conclusion and clinical relevance  Use of the wound soaker catheter was a viable means of providing local analgesia in post-operative veterinary patients. Studies are needed to evaluate efficacy of pain management, and to further investigate techniques for catheter placement and maintenance which may help to optimize the analgesia achieved using this technique.     

Inoculation of pearl millet with the downy mildew pathogen, Sclerospora graminicola: chilling inoculum to delay zoospore release and avoid spray damage to zoospores

Inoculation of plants by spraying with a suspension of zoospores (released from sporangia of Sclerospora graminicola ) induced immediate encystment of the zoospores and drastically reduced their ability to cause disease. The incidence of infection in spray-inoculated pearl millet seedlings was reduced from 90% when sporangia were sprayed, to less than 30% when zoospores were sprayed. Rapid encystment was observed when zoospores were sprayed from both a hand-pumped sprayer and a compressed-gas sprayer and was probably caused by shearing forces. Chilling suspensions of sporangia prior to spraying delayed zoospore release and was an effective method for maintaining infection potential. Disease incidence was higher when sporangia were chilled to 0·2°C rather than to 4°C. Chilling resulted in some abnormal zoospore structures being released from sporangia when suspensions were returned to 20°C. The frequency of these structures increased with increasing chilling time. For seedlings sprayed with sporangia before zoospore release, a small reduction in disease incidence was observed when chilled inoculum was used, probably due to cold disruption of zoosporogenesis. For large-scale disease resistance screening, this reduction is outweighed by the benefit of a uniform and adequately high disease pressure that can be obtained over many hours using chilled spore suspensions.     

Molecular evidence of tick-transmitted infections in dogs and cats in the United Kingdom

PCR analysis was used to determine the prevalence of tick-transmitted infections in 120 systemically ill dogs and 60 cats recruited over a period of three months from 52 veterinary practices in the UK. The animals had not travelled outside the UK and had one or more of the following clinical criteria: acute or recurrent pyrexia, anaemia and/or thrombocytopenia, polyarthritis/muscle pain, splenomegaly/lymphadenopathy, and intraocular inflammation with systemic signs. Blood samples from the animals were tested for the presence of DNA from Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum by using simple PCR targeting. B. burgdorferi sensu lato was detected in five dogs and two cats, and A. phagocytophilum was detected in one dog and one cat. These results provide the first molecular evidence of naturally occurring B. burgdorferi sensu lato infection in cats in the UK and confirm that A. phagocytophilum infection is present in cats. There were no statistically significant associations between the infections and the clinical signs shown by the dogs and cats.