Non‐fish meal,non‐fish oil diet development for red sea bream,Pagrus major,with plant protein and graded levels of Schizochytrium sp.: Effect on growth and fatty acid composition

A feeding experiment was conducted to determine the optimal formulation level of algae meal, which is rich in docosahexaenoic acid (DHA), in a non‐fish meal diet. Six iso‐nitrogenous (450 g/kg) and iso‐lipidic (130 g/kg) experimental diets were prepared. The control diet was formulated with fish meal (400 g/kg), fish oil (60 g/kg), plant protein sources (220 g/kg) and rapeseed oil (50 g/kg). Plant protein sources (soy protein concentrate, soybean meal and corn gluten meal), rapeseed and fish oil were formulated in the second diet (NFM + FO). In the third diet, fish oil of the NFM + FO diet was replaced by rapeseed oil (NFM + NFO) and designated as the negative control. In the other three diets, rapeseed oil in the NFM + NFO diet was replaced with algae meal (Schizochytrium sp. powder) at 50 g/kg, 100 g/kg and 150 g/kg (AM5, AM10 and AM15, respectively). Triplicate groups of juvenile red sea bream (8.8 g) were fed the experimental diets for 12 weeks near satiation. The growth was lowest in the fish fed NFM + NFO diet. This was improved by the formulation of algae meal, which reached the growth level of the NFM + FO group in the AM10 group. The lipid content of the whole fish body in the NFM + NFO group was significantly lower than those of other groups. The fatty acid profile showed significant differences among dietary treatments. DHA content in total and polar lipids of the whole body and liver was highest in the AM10 and AM15 groups. These results reconfirm that microalgae are a suitable lipid source for the replacement of dietary fish oil for marine fish, and the optimal level was estimated as 50 g/kg?100 g/kg in diet.     

Bispectral index as an indicator of anaesthetic depth during isoflurane anaesthesia in the pig

The objective of the study was to examine the relationship between the ‘depth’ of anaesthesia - as determined by clinical signs - and the bispectral index (BIS). Electroencephalograms (EEG)s were recorded in 8 female and 8 castrated male, healthy Norwegian landrace pigs undergoing isoflurane anaesthesia, from which the bispectral index (BIS) was calculated. Isoflurane was delivered in pure oxygen at end-tidal concentrations of 1.6, 1.9, 2.2 and 2.5%, in randomised order, for 30 min after which the EEG was recorded over a 5 min period. Anaesthetic depth was evaluated on a visual analogue scale (VAS) by an experienced anaesthetist. The 95% confidence interval for the mean correlation coefficient between BIS and VAS was calculated to be -0.52–0.30. Confidence intervals (95%) for the mean change in the BIS obtained during the conscious state and that obtained during anaesthesia at different isoflurane concentrations was also calculated. There was a significant decrease in the BIS recorded during consciousness and after 1.6% isoflurane anaesthesia, and between readings after inhalation of 2.2% and 2.5% isoflurane. This indicates that BIS does not accurately reflect ‘depth’ at surgical levels of isoflurane anaesthesia in the pig, and is of no use for this purpose.     

Survey of mutations of a histidine kinase gene BcOS1 in dicarboximide-resistant field isolates of Botrytis cinerea

Previously, we cloned a putative osmosensing histidine kinase gene (BcOS1) and revealed that a single amino acid substitution, isoleucine to serine at codon 365, conferred dicarboximide resistance in field isolates of Botrytis cinerea. This point mutation (type I) occurred within the restriction enzyme TaqI site of the wild-type BcOS1 gene. Thus, a procedure was developed for detecting the type I mutation of the BcOS1 gene using a polymerase chain reaction (PCR) in combination with restriction fragment-length polymorphism (RFLP). Diagnosis by PCR-RFLP was conducted on the 105 isolates isolated from 26 fields in Japan. All dicarboximide-sensitive isolates (49 isolates) had the wild-type BcOS1 gene, and the 43 isolates with the type I mutation were resistant to dicarboximides without exception. These data indicate that dicarboximide-resistant isolates with type I mutation are widespread throughout Japan. However, other types of dicarboximide resistance were detected among isolates from Osaka; among the 24 resistant isolates from Osaka, 12 had the BcOS1 gene without the type I mutation. BcOS1 gene sequencing of these resistant isolates classified them into two groups, type II and type III. The type II isolates have three amino acid substitutions within BcOS1p (³⁶⁸Val to Phe, ³⁶⁹Gln to His, and ⁴⁴⁷Thr to Ser). The type III isolates have two amino acid substitutions within BcOS1p (³⁶⁹Gln to Pro and ³⁷³Asn to Ser). These amino acid changes are located on the amino acid repeat domain in BcOS1p. The three types of resistant isolates were all moderately resistant to dicarboximides without significant osmotic sensitivity, and their pathogenicity on cucumber leaves was also very similar to that of the wild-type isolate.     

Interactive effects of salinity and complete fishmeal replacement on growth,food consumption,and gene expression of hepatic IGF‐I,IGF‐II and growth hormone receptors in Nile tilapia,Oreochromis niloticus (L.)

A 12‐week feeding trial was conducted using Nile tilapia, Oreochromis niloticus (L.) to evaluate the interactive effects of fishmeal replacement and salinity on growth, feed utilization efficiencies and relative expression of growth related genes. Two iso‐nitrogenous and iso‐energetic diets were prepared (32% protein). The control diet included 15% fishmeal (FM diet) and fishmeal component in non‐fishmeal diet (NFM) was eliminated by a mixture of poultry by‐product meal, high protein distillers dried grains and distillers dried grains with soluble. The NFM diet was supplemented with DL‐methionine and L‐lysine. Duplicated group of fish with initial mean weight of 6 g, reared in four salinity levels (0, 4, 8 and 12 g/L) were fed one of the two diets twice a day to near satiety. At the end of the experiment, growth, feed utilization efficiency and expression of growth related genes were compared. The specific growth rate (SGR), mean feed intake (MFI) and feed conversion ratio (FCR) were not affected by the diets while salinity effects were significant. The fish in the 4 g/L salinity showed the highest SGR and MFI while fish in the 0 g/L treatment showed the lowest FCR. Relative expression of hepatic IGF‐I and IGF‐II was regulated by salinity but not by the diet. Expression of growth hormone receptor gene was not affected by either diet or salinity. The present findings provide evidence for the possibility of total fishmeal replacement in saline waters (0–12 g/L) without compromising growth, feed utilization and body composition of Nile tilapia.     

Characterisation of QoI‐resistant field isolates of Botrytis cinerea from citrus and strawberry

BACKGROUND: In 2004, field isolates of Botrytis cinerea Pers. ex Fr., resistant to strobilurin fungicides (QoIs), were first found in commercial citrus orchards in Wakayama Prefecture, Japan. Subsequently, QoI‐resistant isolates of this fungus were also detected in plastic strawberry greenhouses in Saga, Ibaraki and Chiba prefectures, Japan. Biological and molecular characterisation of resistant isolates was conducted in this study. RESULTS: QoI‐resistant isolates of B. cinerea grew well on PDA plates containing kresoxim‐methyl or azoxystrobin at 1 mg L^?1, supplemented with 1 mM of n‐propyl gallate, an inhibitor of alternative oxidase, whereas the growth of sensitive isolates was strongly suppressed. Results from this in vitro test were in good agreement with those of fungus inoculation tests in vivo. In resistant isolates, the mutation at amino acid position 143 of the cytochrome b gene, known to be the cause of high QoI resistance in various fungal pathogens, was found, but only occasionally. The heteroplasmy of cytochrome b gene was confirmed, and the wild‐type sequence often present in the majority of resistant isolates, indicating that the proportion of mutated cytochrome b gene was very low. CONCLUSION: The conventional RFLP and sequence analyses of PCR‐amplified cytochrome b gene are insufficient for molecular identification of QoI resistance in B. cinerea. Copyright © 2009 Society of Chemical Industry     

Abnormal motor activity during anaesthesia in a dog: a case report

Seizures or convulsions that occur during anaesthesia in veterinary patients are infrequently reported in the literature. Consequently, the incidence of such events is unknown. Several drugs commonly used in clinical veterinary anaesthesia have been shown to induce epileptiform activity in both human clinical patients and experimental candidates. The present case report describes convulsions in a four-year old male Bernese mountain dog during maintenance of anaesthesia with isoflurane after premedication with acepromazine and methadone followed by co-induction with propofol and ketamine. The dog had no history of previous convulsions. The use of several sedative and anaesthetic drugs makes it difficult to find one single causative pharmaceutical.     

Hypercalcemia in a dog with resolution of iatrogenic Cushing's syndrome

A six-year-old spayed Pug was presented with crust formation and ulcer on the skin. The patient had received long-term glucocorticoid therapy for treatment of tentatively diagnosed panniculitis. Severe calcification and pyoderma was observed and the patient was diagnosed with iatrogenic Cushing's syndrome and predonisolone was gradually withdrawn. After the withdrawal, the patient developed marked hypercalcemia (15.3 mg/dl) and finally died from renal failure. It is postulated that the eluted calcium from the calcified lesions may have contributed to the high serum calcium level as the underlying disease was not identified on necropsy.     

Phylogenetic analysis of Fusobacterium necrophorum, Fusobacterium varium and Fusobacterium nucleatum based on gyrB gene sequences

The nucleotide sequences of the DNA gyrase B subunit gene (gyrB) of Fusobacterium necrophorum subsp. necrophorum, F. necrophorum subsp. funduliforme and F. varium were determined and analyzed together with those of F. nucleatum subsp. nucleatum and F. nucleatum subsp. vincentii. On the phylogenetic tree constructed, the strains of each fusobacterial species formed distinct clusters with deep sublines. The degree of sequence similarity within each cluster was 93.2% or more, whereas similarities between clusters ranged from 70.1 to 72.7%. These clusters were recovered with 100% bootstrap probabilities and are in very good agreement with the species of Fusobacterium. These data suggest that gyrB is an accurate genealogical marker for the classification of the fusobacterial taxa considered in this study.