Episodic blindness and ataxia in a horse with cholesterinic granulomas

An 11‐year‐old Oldenburg mare presented following three episodes of acute, transient blindness, ataxia, and disorientation within the preceding 7 months. Clinical improvement, including return of vision, occurred within 1 week of initiating corticosteroid therapy for each of the three episodes. However, mild right‐sided miosis was a consistent finding on ophthalmic examinations. Routine clinicopathologic testing revealed no significant abnormalities, and testing of cerebral spinal fluid for selected infectious diseases was unrewarding. Computed tomography of the brain demonstrated a hyperattenuating mass with peripheral mineralization in the rostroventral aspect of each lateral ventricle. The mare was euthanized due to a guarded to poor prognosis. On histopathology, the masses consisted of clusters of cholesterol clefts admixed with leukocytes, mineral deposits, and connective tissue. Cholesterinic granulomas of the lateral ventricles and hydrocephaly were diagnosed. Cholesterinic granulomas should be considered a differential diagnosis in horses presenting for intermittent blindness.     

Effects of Bacillus subtilis B10 spores on viability and biological functions of murine macrophages

This study was conducted to investigate the effects of Bacillus subtilis B10 spores on the viability and biological functions of murine macrophage. RAW 264.7 cells were stimulated both with and without B. subtilis B10 spores for 12 h. Then cell viability was determined to evaluate the cytotoxic effect of B. subtilis B10 spores to the cells, and the activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH), the production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), and the secretion of inflammatory cytokines were measured to analyze the functions of macrophages. The results showed that B. subtilis B10 spores were not harmful to RAW 264.7 cells and they also strongly enhanced the activities of ACP and LDH (P < 0.01), remarkably increased NO and iNOS production (P < 0.01), and significantly stimulated the secretion of pro‐inflammatory cytokines, including tumor necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin‐1 beta (IL‐1β), IL‐6, IL‐8 and IL‐12 (P < 0.01) while they reduced anti‐inflammatory cytokine IL‐10 (P < 0.01). The outcomes suggest that B. subtilis B10 spores are not only safe for murine macrophages, but also can activate these cells and enhance their immune function. The above findings suggest that B. subtilis B10 spores are potentially probiotic.     

Physiologic and biochemical measurements and response to noxious stimulation at various concentrations of MS‐222 in Koi (Cyprinus carpio)

ObjectiveTo evaluate the physiological effect and response to noxious stimulation at five concentrations of MS-222 in koi (Cyprinus carpio).Study designProspective experimental study.AnimalsTwenty-one healthy adult unknown sex koi fish weighing mean 450 ± SD 120 g.MethodsEach fish was exposed to five different concentrations of MS-222 (50, 70, 110, 150 and 190 mg L^?1) in a random sequence during the same anaesthetic event. For each concentration of MS-222, vital functions such as heart rate (HR) (via Doppler) and opercular rate (OpR) were recorded after a standardized induction period. Response to two noxious stimuli in the form of haemostat clamp pressure applied on the tail and the lip was evaluated, and blood was drawn to measure biochemical and blood gas values.ResultsDecrease in response to noxious stimulation with an increase of MS-222 concentration both for the lip (p = 0.0027) and the tail (p < 0.0001) stimulus was observed. Biochemical values were unaffected by the concentration of MS-222 with the exception of lactate concentration which was weakly correlated with the duration of anaesthesia (r = 0.31, p < 0.001) and the number of times the fish was clamped or bled prior to sampling (r = 0.23, p < 0.001). Opercular rate decreased with the increase in anaesthetic concentration, and HR was not affected.Conclusions and clinical relevanceOur results indicated a decrease in response to stimulus and a decrease in OpR that were associated with increased concentrations of MS-222. This may assist in establishing anaesthetic protocols using MS-222 in fish and supports the use of supramaximal pressure stimuli to teleost fish under variable MS-222 concentrations as a model for future studies.     

Naltrexone does not affect isoflurane minimum alveolar concentration in cats

ObjectiveTo test whether naltrexone, an opioid receptor antagonist, affects the minimum alveolar concentration (MAC) of isoflurane in cats, a species that is relatively resistant to the general anesthetic sparing effects of most opioids.Study designRandomized, crossover, placebo-controlled, blinded experimental design.AnimalsSix healthy adult cats weighing 4.9 ± 0.7 kg.MethodsThe cats were studied twice. In the first study, baseline isoflurane MAC was measured in duplicate. The drug (saline control or 0.6 mg kg^?1 naltrexone) was administered IV every 40–60 minutes, and isoflurane MAC was re-measured. In the second study, cats received the second drug treatment using identical methods 2 weeks later.ResultsIsoflurane MAC was 2.03 ± 0.12% and was unchanged from baseline following saline or naltrexone administration.Conclusion and clinical relevanceMinimum alveolar concentration was unaffected by naltrexone. Because MAC in cats is unaffected by at least some mu-opioid agonists and antagonists, spinal neurons that are directly modulated by mu-opioid receptors in this species cannot be the neuroanatomic sites responsible for immobility from inhaled anesthetics.     

Evaluation of electrical nerve stimulation for epidural catheter positioning in the dog

ObjectiveTo evaluate the accuracy of epidural catheter placement at different levels of the spinal cord guided solely by electrical nerve stimulation and resultant segmental muscle contraction.Study designProspective, experiment.AnimalsSix male and two female Beagles, age (1 ± 0.17 years) and weight (12.9 ± 1.1 kg).MethodsAnimals were anesthetized with propofol and maintained with isoflurane. An insulated epidural needle was used to reach the lumbosacral epidural space. A Tsui epidural catheter was inserted and connected to a nerve stimulator (1.0 mA, 0.1 ms, 2 Hz) to assess positioning of the tip at specific spinal cord segments. The catheter was advanced to three different levels of the spinal cord: lumbar (L2–L5), thoracic (T5–T10) and cervical (C4–C6). Subcutaneous needles were previously placed at these spinal levels and the catheter was advanced to match the needle location, guided only by corresponding muscle contractions. Catheter position was verified by fluoroscopy. If catheter tip and needle were at the same vertebral body a score of zero was assigned. When catheter tip was cranial or caudal to the needle, positive or negative numbers, respectively, corresponding to the number of vertebrae between them, were assigned. The mean and standard deviation of the number of vertebrae between catheter tip and needle were calculated to assess accuracy. Results are given as mean ± SD.ResultsThe catheter position in relation to the needle was within 0.3 ± 2.0 vertebral bodies. Positive predictive values (PPV) were 57%, 83% and 71% for lumbar, thoracic and cervical regions respectively. Overall PPV was 70%. No significant difference in PPV among regions was found.Conclusion and clinical relevancePlacement of an epidural catheter at specific spinal levels using electrical nerve stimulation was feasible without radiographic assistance in dogs. Two vertebral bodies difference from the target site may be clinically acceptable when performing segmental epidural regional anesthesia.     

Saw Palmetto (Serenoa repens) Flowering and Fruiting Response to Time Since Fire

Saw palmetto (Serenoa repens [Bartr.] Small) is a shrubby palm common in southeastern US pine flatwoods ecosystems. Demand recently has increased for fruits for the herbal remedies market. Because only wild saw palmettos are harvested, management strategies are needed to promote flowering and fruiting. This study investigated effects of time since growing season (April–July) fires on flowering and fruiting of saw palmetto ramets ≥ 54 cm in height, in 18 pine flatwoods or dry prairie sites (six sites in three locations, burned in 1996, 1995, 1994, 1993, 1992, or before 1991) in central and southwest Florida from 1996 to 1999. We used repeated measures, linear mixed models to test for time since fire effects on proportion of ramets flowering, proportion of ramets fruiting, and fruit yield. Ranges of means among sites over all years of the study for proportion of ramets flowering, proportion of ramets fruiting, and fruit yield were 0 to 0.78, 0 to 0.72, and 0 kg · ha^?1 to 2 869 kg · ha^?1, respectively. Time since fire strongly influenced flowering; highest probability of flowering occurred 1 yr after burning, followed by an abrupt decrease 2 yr after burning, then a gradual increase from 3 to 5 yr after fires (polynomial regression, P < 0.0001 for fixed effects). Probability of fruiting increased with increasing time since fire (quadratic regression, P < 0.001 for fixed effects), but fruit yields showed no pattern in response to time since fire (P=0.916). The decrease in influence of fire from flowering through fruit maturity presumably was caused by mortality from factors such as caterpillar predation and fungal infection. To promote increased flowering and fruit yields, we recommend that growing season burns be conducted approximately every 5 yr. We suggest, however, that management strategy be modified as necessary to maintain ecosystem diversity and function.     

The use of T‐DNA tagging to isolate mutants of Colletotrichum gloeosporioides and Colletotrichum acutatum with reduced virulence against Hevea brasiliensis

Agrobacterium tumefaciens‐mediated transformation (ATMT) is being increasingly recognized as an effective insertional mutagenesis tool in studies of filamentous fungi for gene discovery and functional analysis. We developed and optimized ATMT for 2 Colletotrichum species, Colletotrichum gloeosporioides and Colletotrichum acutatum, the causative agents of Colletotrichum leaf disease in rubber trees in Southern China. A. tumefaciens strain AGL‐1 carrying an ILV1 gene and a green fluorescent protein gene were used to transform the conidia of these 2 Colletotrichum species. The transformation efficiency was correlated with the co‐cultivation duration and bacterial cell concentrations, which reached 300–400 transformants per 1 × 10⁶ conidia after optimization. Southern blot analysis indicated that about 60.0% of the C. gloeosporioides transformants and 46.2% of the C. acutatum transformants had a single copy of T‐DNA in their genomes. Fungal genomic DNA segments flanking the T‐DNA were identified in the transformants through thermal asymmetrical interlaced polymerase chain reaction followed by sequencing. The flanking sequences from 4 C. acutatum and 7 C. gloeosporioides transformants showed moderate or weak homology to the NCBI database entries. Some sequences matching those reported virulence‐related genes. The results suggest that the T‐DNA inserted mutants banks constructed are useful for the discovery of new or important genes and to elucidate their function in the 2 Colletotrichum species.     

Western corn rootworm egg hatch and larval development under constant and varying temperatures

Predicting western corn rootworm (Diabrotica virgifera virgifera LeConte; WCR) development in the field depends on models that use experimentally determined degree-days (°D). For constant temperature regimes, this temperature sum can be reliably used to predict hatch and development of WCR larvae. In the first experiment in climate cabinets, we evaluated the effects of varying day–night (4 and 6 °C difference) temperature regimes compared to equivalent constant temperature regimes on hatch, development, and recovery of WCR larvae from a non-diapausing population. Relative to constant diurnal temperatures, varying day–night temperatures resulted in earlier larval hatch and accelerated larval development (especially when day–night temperatures differed by 6 °C) due to direct temperature effects (i.e., the Kaufmann-effect) and to enhanced plant growth. For WCR eggs, the temperature sums needed for hatch in the field are overestimated when they are determined by models based on constant experimental temperatures. Recovery of larvae from soil was not affected by temperature, but was positively associated with plant height. In a second experiment we evaluated whether the found effect of varying temperature ranges on the acceleration of larval hatch is also influenced by the level of these varying temperatures. Initial hatch started earlier by varying diurnal temperatures only under a low-temperature regime (14 ± 4 °C). For herbivorous insects like WCR, plant growth effects induced by varying temperature regimes may result in increased variation in temperature-based developmental parameters. Consideration of these effects will improve models that predict WCR hatching patterns and will improve the development and timing of control strategies.