Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Spirochetemia caused by Borrelia turicatae infection in 3 dogs in Texas

Spirochetemia was diagnosed in 2 Siberian Huskies and a Rottweiler from the northwestern region of Texas between June 1999 and October 2001. Clinical findings were nonspecific; tick exposure was documented in 2 of the dogs. Hematologic abnormalities included anemia (n=2), neutrophilia (n=2, including 1 with a left shift), lymphopenia (n=3), eosinopenia (n=3), and thrombocytopenia (n=2). One anemic dog had a positive Coombs' test. In 1 dog, Western blot analysis of serum yielded multiple positive bands with B turicatae lysate, indicating the spirochetemia most likely was due to B turicatae infection. In 2 dogs, spirochetes were cultured from the blood and identified using DNA analysis as Borrelia turicatae; 1 of these dogs also was seropositive for Ehrlichia canis and B burgdorferi. In 2 cases, spirochetemia was more prominent in blood smears prepared immediately after sample collection than in smears prepared from EDTA blood. Two dogs recovered with doxycycline treatment; 1 dog declined clinically despite treatment and was euthanized. B turicatae is the agent of tick-borne (endemic) relapsing fever in humans and is distinct from B burgdorferi, the agent of Lyme disease; however, serologic cross-reactivity may occur. B turicatae is transmitted by the soft tick, Ornithodoros turicata, and infection should be considered in dogs with spirochetemia and possible exposure to the tick vector.     

Effects of reconcile (fluoxetine) chewable tablets plus behavior management for canine separation anxiety

Canine separation anxiety is a common behavioral problem presented to veterinarians. Associated behaviors are distressing to both dog and owner, have the potential to disrupt the human-companion animal bond, and may lead to euthanasia. The results of this study demonstrate the clinical efficacy and safety of Reconcile (fluoxetine, 1 to 2 mg/kg/day [0.45 to 0.91 mg/lb/day]), in conjunction with behavior management, for the treatment of canine separation anxiety. The beef flavored chewable formulation was palatable to treated dogs and easy to administer. This study provides to veterinarians and owners valuable information about an effective separation anxiety treatment plan that combines use of Reconcile with behavior modification.     

Dietary zinc concentration and lipopolysaccharide injection affect circulating trace minerals,acute phase protein response,and behavior as evaluated by an ear-tag–based accelerometer in beef steers

To assess plasma trace mineral (TM) concentrations, the acute phase protein response, and behavior in response to a lipopolysaccharide (LPS) challenge, 96 Angus cross steers (average initial body weight [BW]: 285 ± 14.4 kg) were sorted into two groups by BW (heavy and light; n = 48/group), fitted with an ear-tag–based accelerometer (CowManager SensOor; Agis, Harmelen, Netherlands), and stagger started 14 d apart. Consecutive day BW was recorded to start the 24-d trial (days −1 and 0). Dietary treatments began on day 0: common diet with either 30 (Zn30) or 100 (Zn100) mg supplemental Zn/kg DM (ZnSO₄). On day 17, steers received one of the following injection treatments intravenously to complete the 2 × 3 factorial: 1) SALINE (~2–3 mL of physiological saline), 2) LOWLPS: 0.25 µg LPS/kg BW, or 3) HIGHLPS: 0.375 µg LPS/kg BW. Blood, rectal temperature (RT), and BW were recorded on day 16 (−24 h relative to injection), and BW was used to assign injection treatment. Approximately 6, 24 (day 18), and 48 (day 19) h after treatment, BW, RT, and blood were collected, and final BW recorded on day 24. Data were analyzed in Proc Mixed of SAS with fixed effects of diet, injection, diet × injection; for BW, RT, dry matter intake (DMI), plasma TM, and haptoglobin-repeated measures analysis were used to evaluate effects over time. Area under the curve analysis determined by GraphPad Prism was used for analysis of accelerometer data. Body weight was unaffected by diet or injection (P ≥ 0.16), but there was an injection × time effect for DMI and RT (P < 0.05), where DMI decreased in both LPS treatments on day 16, but recovered by day 17, and RT was increased in LPS treatments 6 h post-injection. Steers receiving LPS spent less time highly active and eating than SALINE (P < 0.01). Steers in HIGHLPS spent lesser time ruminating, followed by LOWLPS and then SALINE (P < 0.001). An injection × time effect (P < 0.001) for plasma Zn showed decreased concentrations within 6 h of injection and remained decreased through 24 h before recovering by 48 h. A tendency for a diet × time effect (P = 0.06) on plasma Zn suggests plasma Zn repletion occurred at a greater rate in Zn100 compared to Zn30. These results suggest that increased supplemental Zn may alter the rate of recovery of Zn status from an acute inflammatory event. Additionally, ear-tag–based accelerometers used in this study were effective at detecting sickness behavior in feedlot steers, and rumination may be more sensitive than other variables.     

Potential for monoterpenes to affect ectomycorrhizal and saprotrophic fungal activity in coniferous forests is revealed by novel experimental system

The fungal community in coniferous forest soils plays a pivotal role in ecosystem processes such as decomposition and carbon and nutrient cycling. Both saprotrophic (SP) and ectomycorrhizal (ECM) fungi occur throughout the upper soil horizons in coniferous forests and could therefore be exposed to high concentrations of monoterpenes occurring in the needle litter and roots of some tree species. Previous work has noted the differential effects of monoterpenes on the mycelial growth of a range of both SP and ECM fungi when grown in artificial nutrient media. This study used a novel experimental system to assess the effect of environmentally relevant concentrations of monoterpenes on the activity of ECM and SP fungi grown on more natural substrata. Exposure of the ECM fungus Paxillus involutus (Batsch) Fr. to vapours of either α-pinene or β-pinene resulted in a significantly greater proportion of root tips being colonised by the fungus when it was grown with seedlings of Picea abies (L.) Karst. Exposure to monoterpenes resulted in a significant decrease in respiration rate of two species of litter degrading SP fungi, Mycena galopus var. candida J. E. Lange and Collybia butyracea (Bull.) P. Kumm. There was no difference in response between the two SP species, despite the fact that previous tests in liquid nutrient media, with monoterpenes at higher concentrations, indicated that one species was sensitive and one was not. The high volatility and low solubility of monoterpenes in water make them challenging to work with. The experimental system developed here, although still artificial, provides a bridge between pure culture studies in defined media and all the complexities of forest soils in the field, by allowing the exposure of fungi to environmentally relevant monoterpene concentrations in more natural substrata.     

Vincristine overdose in a cat: clinical management, use of calcium folinate, and pathological lesions

A 6-year-old female neutered Burmese cat received a 10 times overdose (5mg/m(2)) of vincristine, administered in error. Supportive therapy, including administration of calcium folinate, was instigated within 8h. Despite treatment, the patient exhibited deterioration in renal and respiratory function and died 72 h after overdose. Necropsy was performed within 24h of death. Gross examination revealed pulmonary oedema and a pale brown liver with a prominent lobular pattern. Histological examination revealed marked apoptosis and necrosis of the bone marrow myeloid series, and mild to moderate apoptosis and necrosis of the erythroid and megakaryocyte series. Multifocal necrosis of the renal tubules, hepatocytes, and small intestinal crypt epithelium was also observed. Use of calcium folinate as a rescue therapy following vincristine overdose in humans has been previously documented. If treatment is to be successful in cases of vincristine overdose in cats, then a more complete understanding of the pathogenesis of vincristine toxicity in companion animal species is required.     

Clinical and magnetic resonance imaging findings in 92 cats with clinical signs of spinal cord disease

Medical records of 92 cats presented with clinical signs of spinal cord disease, which had undergone magnetic resonance imaging (MRI), were reviewed. The cats were grouped into seven categories based upon the diagnosis suggested by results of MRI, cerebrospinal fluid analysis and other diagnostic procedures: neoplastic (n=25), inflammatory or infectious (n=13), traumatic (n=8), vascular (n=6), degenerative (n=5), anomalous (n=3) and those with an unremarkable MRI (n=32). There were two independent predictors of abnormal MRI findings: severity of clinical signs and presence of spinal pain. Abnormal MRI findings and speed of onset of disease were significantly associated with survival. For the 32 cats with unremarkable MRI findings, only nine died due to spinal disease and, therefore, the median survival time (MST) was not reached (lower 95% confidence interval (CI)=970 days). For the 60 cats with abnormal MRI findings, 37 died due to their disease and the MST was 138 days (95% CI: 7-807).     

Effect of curing at different temperatures on biochemical composition of onion (Allium cepa L.) skin from three freshly cured and cold stored UK-grown onion cultivars

Onions are cured in order to form a complete, dry, outer skin which reduces water loss and suppresses incidence of disease, and can promote a darker skin finish. Currently in the UK, standard curing practises for onions involves heating at 28 °C for six weeks (65–75% RH), however, reducing curing temperatures may help to reduce energy usage. There is little empirical data on the effects of curing temperature on flavonol concentration in the skin of brown onions and on flavonol and anthocyanin concentration in the skin of red onions. Therefore, the aim of this study was to elucidate the compounds responsible for the change in onion skin colour when cured at different temperatures.Brown cvs. Sherpa and Wellington, and red onions cv. Red Baron, were cured at 20, 24 or 28 °C for six weeks. Replicated skin samples were analysed immediately after curing and after seven months cold storage at 1 ± 0.5 °C. Measurement of objective colour showed that skin of cvs. Sherpa and Wellington was darker and had a lower hue angle (H°) immediately after being cured at 28 °C compared to 20 °C. In contrast, skin of cv. Red Baron had a higher H° but no change in lightness (L*) when cured at 28 °C compared to 20 °C. Fructose, sucrose and glucose concentrations were analysed as they are thought to play a role in regulating the synthesis of flavonols and anthocyanins, both coloured compounds found in onion skin; however no significant correlations were found between colour data and sugar concentrations. Flavonols were measured in the skin of all cvs. and anthocyanins in the skin of cv. Red Baron. Quercetin glucoside and anthocyanin concentrations in the skin of onions cv. Red Baron immediately after curing were higher in those cured at 20 °C. Total flavonols and total anthocyanins were negatively correlated with H° in the skin of onions cv. Red Baron, but there was no similar correlation between total flavonols and H° for onion cvs. Sherpa and Wellington. This suggests that anthocyanins and flavonols may play a major role in varying skin colour of red onions cv. Red Baron cured at different temperatures; however, the difference between curing temperatures may not have been sufficient to represent a correlation between darkening of cvs. Sherpa and Wellington and flavonol concentration. Further investigation is therefore required to fully elucidate the compounds responsible for colour changes observed in brown onions.     

The physical action of three diatomaceous earths against the cuticle of the flour mite Acarus siro L. (Acari: Acaridae)

Experiments examined the accepted modes of action of the desiccant dust, diatomaceous earth (DE), against the flour mite, Acarus siro (L.) (Astigmata). Adult mites were exposed to three DE products for periods of 3, 18 and 72 h under conditions designed to allow partial desiccation of the mites without causing mortality. After exposure, the DE dust particles were washed off the mite bodies, and both the DE and the mites were examined for presence of cuticular hydrocarbons by gas chromatography-mass spectroscopy (GC-MS) analysis. GC-MS identified seven major cuticular lipids of chain length C(13)--C(26) that may have a role in the waterproofing of A. siro. After 18 h exposure, n-tridecane and several different long-chain fatty acid ethyl esters were detected on one of the DE products. After 72 h, n-tridecane was detected on all three DE products. Mite samples retained after removal of the DEs were examined by microscopy. Tentative evidence was observed by conventional low-power microscopy that might indicate uniform removal of the epicuticle. However, a detailed examination by scanning electron microscopy showed no signs of abrasion.     

Evaluation of corn plant as potential banker plant for supporting predatory gall midge, Feltiella acarisuga (Diptera: Cecidomyiidae) against Tetranychus urticae (Acari: Tetranychidae) in greenhouse vegetable production

The twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important and highly polyphagous pests of vegetables and other crops worldwide. Experiments were conducted in the laboratory and greenhouse to evaluate corn (Zea mays L.) as a banker plant for the predatory gall midge, Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae) to potentially control T. urticae. Choice and no-choice experiments were carried out to determine the host plant preference of an alternative prey, Oligonychus pratensis (Banks)(Acari: Tetranychidae) to corn and green bean (Phaseolus vulgaris L.). Results showed that O. pratensis adults strongly preferred corn as a host plant and posed no risk to green bean. F. acarisuga was found to fly at least 7.0 m to search for new preys on green bean plants, and over 176 F. acarisuga larvae per leaf were recorded at 14 d after dispersal. F. acarisuga proved to be an excellent predator of both T. urticae and O. pratensis. The predation by F. acarisuga to T. urticae and O. pratensis ranged from 43.7 to 67.9% and 59.2 to 90.3%, respectively, under laboratory conditions. In a non-cage study, 81.2% of T. urticae population was suppressed by F. acarisuga in reference to the control (cage treatment). The results showed that this banker plant system has potential for controlling T. urticae in greenhouse vegetable production.