Diagnosis of canine Hepatozoon spp. infection by quantitative PCR

Hepatozoon (H.) americanum and H. canis are the etiological agents of canine hepatozoonosis, a disease that is found worldwide and is also prevalent in the southeastern United States. Current laboratory diagnosis of canine hepatozoonosis caused by H. americanum is usually dependent on visual identification of Hepatozoon "onion skin cysts" in muscle biopsies, an approach that requires invasive sampling and can result in false negatives. We have developed a diagnostic method for detection of Hepatozoon spp. DNA that integrates nucleic acid extraction with extensive agitation to maximize DNA extraction efficiency. The DNA extracted from canine EDTA-whole blood is subjected to real-time PCR, and fluorescence resonance energy transfer (FRET) probes detect a signature polymorphism in the amplified DNA. This PCR method amplifies a fragment of the Hepatozoon 18S rDNA gene, detects as few as 7 genomic copies of Hepatozoon spp. per ml of blood with high specificity, and differentiates between H. americanum and H. canis amplicons. A surprising 300-fold increase of H. americanum 18S rDNA targets occurred during 3-0 days of storage of positive blood specimens. Examination of 614 EDTA-blood samples submitted mostly from the southeastern Unites States from dogs with suspected hepatozoonosis identified H. americanum in 167 samples (27.2%). An additional 14 samples (2.3%) were positive for H. canis, and 14 samples (2.3%) were positive for both H. americanum and H. canis. These results suggest that the Hepatozoon spp. 18S rDNA quantitative PCR may be a valuable tool that can improve diagnosis and therapy of canine hepatozoonosis.     

First report of a mecA-positive multidrug-resistant Staphylococcus pseudintermedius isolated from a dog in New Zealand

CASE HISTORY: A 14-year-old neutered male Sealyham terrier was referred for assessment of a persistent pyoderma. It had experienced numerous episodes of dermatitis involving pododermatitis, pyoderma and otitis over the previous 6 years.

CLINICAL FINDINGS: Superficial, focally deep and mucocutaneous pyoderma were present, with yellow mucoid exudate on both nares and the lower lips crusted with haemopurulent exudate. Epidermal collarettes were present on the dorsal and lateral trunk. There were peri-anal crusts and mild erythema was present on the concave aspect of both pinnae.

MICROBIOLOGICAL FINDINGS: Culture and microbiological testing identified Staphylococcus pseudintermedius as the infecting organism. Kirby-Bauer disc susceptibility testing revealed the isolate was resistant to numerous antimicrobials including oxacillin. PCR testing of the isolate identified the presence of the mecA gene which confers resistance to β-lactam antimicrobials. Pulsed field gel electrophoresis typing suggested the isolate was not related to the methicillin-resistant S. pseudintermedius that had been reported to be associated with canine infections in Western Australia.

DIAGNOSIS: Superficial, deep and mucus membrane pyoderma associated with a multi-drug resistant S. pseudintermedius.

CLINICAL RELEVANCE: This is the first recorded case of canine pyoderma involving methicillin-resistant multidrug-resistant S. pseudintermedius in New Zealand. Treatment of such cases is difficult because the number of effective and available antimicrobials is limited. This finding should raise the awareness of the veterinary and medical professions to the presence of such organisms in New Zealand and stimulate a discussion about possible biosecurity barriers, treatment strategies and prevention of zoonotic and nosocomial infections.     

Light transmissivity of tube shelters affects root growth and biomass allocation of Quercus ilex L. and Pinus halepensis Mill

Context

Tube shelters have been shown to enhance field performance of several Mediterranean species, but responses of newly planted seedlings to the microenvironment induced by shelter walls with different light transmissivity are still poorly documented.

Aims

We studied effects of a range of shelters with varying light transmissivity on post-planting seedling responses during the wet season establishment phase for two Mediterranean trees of contrasting functional ecology.

Methods

Root growth, biomass allocation, water potential, and chlorophyll fluorescence of Quercus ilex and Pinus halepensis seedlings were evaluated across shelters varying in light transmissivity (80, 40, 20, and 10 % plus a mesh shelter) with irrigation.

Results

Plants in dark tubes (20 and 10 % light transmissivity) had less above- and belowground growth and more than two times greater leaf to protruding roots mass ratio, with shoot growth response of Q. ilex being less plastic. Ratio of leaf area/protruding roots area decreased when light transmissivity increased, although no differences were found at ≥40 % transmissivity. Xylem water potential indicated lack of water stress, and high maximum photosynthetic efficiency (F _v/F _m) values show no photoinhibition symptoms irrespective of light transmissivity.

Conclusion

Shelter transmissivity ≥40 % promotes rapid and vigorous root growth immediately after planting for these species. This minimum transmissivity should be considered as a target when designing shelters to help root development and improve water balance of Mediterranean seedlings.     

Comparative Oxygen Consumption and Metabolism of Striped Bass Morone saxatilis and its Hybrid M. chrysops♀ xM. saxatilis♂

It has been reported that metabolic rates of striped bass Morone saxatilis and hybrid striped bass M. chrysops♀ x M. saxatilis♂ are different. A series of experiments were conducted to further characterize oxygen consumption and metabolism of striped bass and its hybrid, the sunshine bass. Oxygen consumption was measured to determine standard and routine metabolic rates of striped bass and hybrid striped bass in a freshwater, flow‐through tank system. Additionally, blood chemistry stress indicators of the two bass groups were compared in both fresh and brackish water. Hematocrit (% PCV) and hemoglobin were measured in order to compare oxidative efficiencies of the bass. Plasma glucose, chlorides, and cortisol levels were measured to compare the relative stress status of the two bass types reared under experimental conditions. No significant differences were found in average daily oxygen consumption between striped bass and sunshine bass for either standard metabolism (P= 0.92), or routine metabolism (P = 0.86). Standard metabolic rates of oxygen consumption were 69 ± 4.1 and 68 ± 3.5 mg 0₂/kg³/⁴ bw/h for sunshine bass and striped bass respectively. Routine metabolic rates were 132 ± 30 and 125 ± 30 mg O₂/kg^3/4 bw/h for sunshine bass and striped bass respectively. While there were no significant differences in oxygen consumption between species, normal feeding activity generally resulted in increased oxygen consumption by the fish. Striped bass had significantly lower hematocrit values (P= 0.0001), but significantly higher hemoglobin concentrations than sunshine bass maintained in freshwater (P= 0.0001). Striped bass had significantly higher (P= 0.0001) levels of plasma glucose compared to sunshine bass (176 ± 8.6 vs. 103 ± 5.6 mg/dL respectively). Plasma chloride levels for striped bass (123 ± 1.9 mEq/L) were significantly higher (P= 0.041) than plasma chloride levels of sunshine bass (117 ± 1.7 mEq/L). Plasma cortisol levels were significantly higher (P= 0.0081) for striped bass (147 ± 8.4 ng/mL) compared to sunshine bass (119 ± 5.6 ng/ mL) when reared in freshwater. When maintained in brackish water, sunshine bass had significantly higher hematocrit values (P= 0.0001), and hemoglobin concentrations (P= 0.0012) when compared to striped bass. However, sunshine bass had significantly higher hemoglobin concentrations (P= 0.0012) when compared to striped bass. In addition, plasma glucose levels were significantly lower (P = 0.0079) for sunshine bass (79 ± 4.1 g/dL) when compared to striped bass (115 ± 11 g/dL). There were no significant differences between the bass in levels of chlorides or cortisol. No differences were detected in oxygen consumption. However, hybrid striped bass may have more efficient oxidative metabolism due to elevated hemoglobin concentrations. While striped bass hemoglobin values tended to be higher in brackish water than in freshwater, sunshine bass hematocrit or hemoglobin values generally were significantly higher than striped bass in both fresh and brackish water. Based on these results, hybrid striped bass may be capable of directing more energy towards growth than striped bass due to more efficient oxidative metabolism and lower losses of energy related to increased stress.     

Treatment of primary immune‐mediated hemolytic anemia with mycophenolate mofetil in two cats

Objective – To describe the use of oral mycophenolate mofetil (MMF) as an adjunctive therapy in 2 cats with primary immune‐mediated hemolytic anemia. Case Series Summary – Two cats suffering from presumptive primary immune mediated hemolytic were treated with MMF as part of their treatment regimens. Both cats had improved complete blood counts following therapy. New or Unique Information Provided – This is the first reported use of oral MMF as adjunctive treatment for cats with immune‐mediated hemolytic anemia. Outcome was favorable in both cats and no adverse effects were noted from the MMF.     

Photosynthetic assimilation and carbohydrate allocation of <Emphasis Type="Italic">Quercus rubra</Emphasis> seedlings in response to simulated herbivory

Introduction  

The artificial regeneration of oaks (Quercus spp.) can supplement deficient natural regeneration, yet growth and physiology of newly planted seedlings are often negatively affected by animal herbivory. Alternative nursery stock types with improved stress resistance may help improve regeneration success.     

Walnut (Juglans spp.) ecophysiology in response to environmental stresses and potential acclimation to climate change

? Context

Walnuts (Juglans spp.) are ecologically and commercially important trees, yet synthesis of past and current research findings on walnut ecophysiology is lacking, especially in terms of potential acclimation to climate change.

? Aims

This study aims to (1) investigate walnut ecophysiology by comparing its attributes to associated deciduous angiosperms, (2) address potential acclimation of walnut to climate change, and (3) identify areas for prioritization in future research.

? Results

There is considerable uncertainty regarding the magnitude of potential effects of climate change on walnut. Some studies tend to indicate walnut could be negatively impacted by climate change, while others do not. Walnut may be at a disadvantage due to its susceptibility to drought and frost injury in current growing regions given the projected increases in temperature and extreme climatic events. Other regions that are currently considered cold for walnut growth may see increased establishment and growth depending upon the rate of temperature increase and the frequency and severity of extreme climatic events.

? Conclusion

Research investigating a combination of environmental factors, such as temperature, carbon dioxide, ozone, water, and nitrogen is needed to (1) better project climate change effects on walnut and (2) develop management strategies for walnut acclimation and adaptation to climate change.     

Growth,Nutrition, and Photosynthetic Response of Black Walnut to Varying Nitrogen Sources and Rates

ABSTRACT

Black walnut (Juglans nigra L.) half-sib 1+0 seedlings were exponentially fertilized with ammonium (NH₄ ⁺) as ammonium sulfate [(NH₄)₂SO₄], nitrate (NO₃ ^?) as sodium nitrate (NaNO₃), or a mixed nitrogen (N) source as ammonium nitrate (NH₄NO₃) at the rate of 0, 800, or 1600 mg N plant^?1 and grown for three months. One month following the final fertilization, N concentration, growth, and photosynthetic characteristics were assessed. Compared with unfertilized seedlings, N addition increased plant component N content, chlorophyll content, and photosynthetic gas exchange. Net photosynthesis ranged from 2.45 to 4.84 μmol m^?2 s^?1 for lower leaves but varied from 5.95 to 9.06 μmol m^?2 s^?1 for upper leaves. Plants responded more favorably to NH₄NO₃ than sole NH₄ ⁺ or NO₃ ^? fertilizers. These results suggest that N fertilization can be used to promote net photosynthesis as well as increase N storage in black walnut seedlings. The NH₄NO₃ appears to be the preferred N source to promote black walnut growth and physiology.