A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B₁₂-responsiveness in lambs and critique existing serum vitamin B₁₂ reference ranges.

METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B₁₂ and MMA, and liveweight gain.

RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 µmol/L when serum vitamin B₁₂ concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 µmol/L) unless serum vitamin B₁₂ concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 µmol/L).

CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B₁₂ are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B₁₂ concentrations. Serum MMA concentrations <13 µmol/L indicate responsiveness to supplementation whilst concentrations <7 µmol/L indicate unresponsiveness. In the range 7–13 µmol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable.

CLINICAL RELEVANCE: The current reference ranges for vitamin B₁₂ responsiveness are conservatively high and lead to over-diagnosis of vitamin B₁₂ deficiency in ill-thriftiness of sheep.     

Seasonal changes in properties of abandoned terraced paddy field soil incubated under different water content conditions

The chemical properties of soil samples collected in August and November from an abandoned terraced paddy field dominated by reeds were examined by in vitro incubation under normal moisture and flooded conditions. Soil pH extracted with water [pH(H₂O)] was higher in soil samples collected from a depth of 0–10 cm in November than in samples collected in August; a high pH(H₂O) was maintained even during nitrification under normal moisture conditions. When soil samples collected in August from a depth of 0–10 cm were incubated under flooded conditions, a significant decrease in reduction potentials (Eh) and an increase in total Fe²⁺ concentrations were observed. Reductive conditions during sampling were strong in soil samples collected in August from a depth of 40–50 cm. Moreover, under normal moisture conditions, soil samples collected in August showed significant decreases in pH(H₂O) and significant production of water-soluble SO₄ ^2? than those collected in November. Glucose addition to soil samples collected from a depth of 0–10 cm caused nitrogen immobilization under normal moisture conditions, increases in exchangeable Fe²⁺ and Mn²⁺, and decreases in exchangeable bases (Ca²⁺, Mg²⁺, K⁺, and Na⁺) under flooded conditions. Seasonal changes in soil properties were probably due to microbial activity and vegetation phenology; thus, the timing of soil sampling influenced incubation experiment results. When abandoned terraced paddy fields are created as biotopes, seasonal changes in reductive soil conditions and slope position must be considered to prevent soil acidification and base cation elution.     

The effect of medetomidine and its antagonism with atipamezole on stress-related hormones, metabolites, physiologic responses, sedation, and analgesia in goats

Six 3‐year‐old goats (three males and three females) weighing 60.0 ± 18 kg (mean ± SD) were used to investigate the effect of medetomidine (MED; 20 µg kg^?1 IV) and its antagonism with atipamezole (ATI; 100 µg kg^?1 IV) on physiologic responses (heart rate (HR; beats minute^?1), respiratory rate (RR; breaths minute^?1), electrocardiogram (ECG), rectal temperature (T; °C), blood pressure (oscillometric; mm Hg), sedation (SED), posture (REC), analgesia (ALG), and stress‐related hormonal and metabolic responses (epinephrine and norepinephrine (high performance liquid chromatography with electrochemical detection), cortisol (COR; µg dL^?1; radioimmunoassay), glucose (GLU; mg mL^?1; enzymatic colorimetric assay), and free fatty acids (modified enzymatic colorimetric assay)); each goat received ATI or SAL in random order separated by 1 week. Jugular catheters were placed for drug administration and blood sampling (10–12 mL sample^?1) using a lidocaine skin block (20 mg) 2 hours prior to beginning of each trial; during this trial, goats breathed room air. Physiologic parameters were measured, SED, REC, and ALG were scored, and blood samples were collected from jugular catheters at baseline (time = ?30 minutes), 5 minutes post‐MED administration (time = ?25 minutes), 25 minute post‐MED administration and immediately prior to antagonism (time = 0 minute), and at 5, 30, 60, and 120 minutes after administering ATI or SAL. ALG was tested by clamping the withers and metacarpus with hoof testers fitted with a force transducer to measure applied isometric force (lb) (a technique used previously in goats to evaluate analgesia). Continuous variables were analyzed by Repeated Measures analysis of variance (anova ); categorical data were analyzed using a Friedman Repeated Measures anova on ranks. A p‐value of <0.05 was considered significant. If a significant difference was found, a Dunnett's pair‐wise comparison of means was conducted. Differences between ATI and SAL were examined at 5, 30, 60, and 120 minutes using a paired t‐test with a Bonferroni correction. Administration of MED resulted in a decrease in T (38.7 ± 0.3 to 34.5 ± 0.4 °C), HR (78 ± 19 to 55 ± 9), and RR (31 ± 12 to 14 ± 5) over time; an increase in mean arterial blood pressure (90 ± 19 to 132 ± 23), COR (0.254 ± 0.125 to 4.327 ± 1.233), and GLU (82.0 ± 13.2 to 255.9 ± 38.9); and changes in SED (alert to marked sedation), REC (standing to recumbent), and ALG (metacarpus = 5 ± 2 to 14 ± 0; withers = 3 ± 2 to 14 ± 0). GLU was 62–70% higher at 60 and 120 minutes and COR was 336% higher after SAL than after ATI at 120 minutes; at 30, 60, and 120 minutes, T was 4–10% higher after ATI than SAL. There were no other significant differences. REC, SED, and ALG were antagonized after ATI. ATI did not antagonize the effect of MED on HR, RR, or MAP, but stabilized T and antagonized the increase in GLU and COR.     

Morphological and molecular characterization for a Japanese isolate of tomato powdery mildew Oidium neolycopersici and its host range

 A single conidium of tomato powdery mildew was isolated from heavily infected leaves of tomato (cv. Moneymaker) grown in the greenhouse of Kinki University, Nara Prefecture, Japan. It was successively multiplied so the morphological and taxonomic characteristics of the pathogen and its host range under high humidity conditions could be analyzed. The isolate KTP-01 of the tomato powdery mildew optimally developed infection structures at 25°C under continuous illumination of 3500 lx. More than 90% of the conidia germinated and developed moderately lobed appressoria. After forming haustoria, the pathogen elongated secondary hyphae from both appressoria and conidia. The hyphae attached to leaf surfaces by several pairs of appressoria and produced conidiophores with noncatenated conidia. In addition to its morphological similarity to Oidium neolycopersici, the phylogenetic analysis (based on the sequence of internal transcribed spacer regions of rDNA) revealed that KTP-01 could be classified into the same cluster group as O. neolycopersici. In host range studies, KTP-01 produced abundant conidia on the foliage of all tomato cultivars tested and tobacco (Nicotiana tabacum), and it developed faint colonies accompanied by necrosis on leaves of potato (Solanum tuberosum), red pepper (Capsicum annuum), petunia (Petunia × hybrida), and eggplant (S. melongena). The pathogen did not infect other plant species including Cucurubitaceae plants, which have been reported to be susceptible to some foreign isolates. Thus, the present isolate of the tomato powdery mildew was assigned as O. neolycopersici, a pathotype different from foreign isolates of the pathogen. Received: December 5, 2002 / Accepted: December 26, 2002 Acknowledgments This work was supported in part by a Grant-in-Aid (12660050) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. We express our deepest thanks to professor Dr. Y. Sato, Toyama Prefectural University, for his kind and valuable suggestion on taxonomic analysis of the powdery mildew pathogen described in the present study.     

Localised radiotherapy for a ferret with possible anal sac apocrine adenocarcinoma

A seven-year-old, neutered male ferret was referred to our hospital with two perianal masses (2.4x3.0 and 2.4x3.5 cm, respectively) that had recurred after initial surgical excision. Complete resection of the masses was impossible as there was deep invasion along the rectum. On histopathology, the masses were diagnosed as apocrine adenocarcinoma possibly of anal gland origin based on tumour location. There was marked response to localised radiotherapy using an orthovoltage unit at 4 Gy, twice weekly. No visible mass was detectable after six doses of radiation. However, at that time, pleural effusion was diagnosed and radiotherapy was discontinued. Cytology of a sample of the pleural effusion suggested mesothelioma, and no obvious pulmonary metastasis of anal sac adenocarcinoma were identified on thoracic radiography. The ferret died at home on day 71 after the first admission.