Immune parameters in the intestine of wild and reared unvaccinated and vaccinated Atlantic salmon (Salmo salar L.)

Forming a barrier to the outside world, the gut mucosa faces the challenge of absorbing nutrients and fluids while initiating immune reactions towards potential pathogens. As a continuation to our previous publication focusing on the regional intestinal morphology in wild caught post smolt and spawning Atlantic salmon, we here investigate selected immune parameters and compare wild, reared unvaccinated and vaccinated post smolts.     

Safety evaluation of a novel muramidase for feed application

Safety evaluation of a muramidase produced by a Trichoderma reesei strain (safe lineage), expressing a muramidase gene isolated from Acremonium alcalophilum is presented. Intended use in feed of this enzyme is as digestive aid in broiler chickens.Muramidase 007, was non-mutagenic and non-clastogenic in vitro, and no adverse effects were observed in 90-day subchronic toxicity studies in rats at doses up to 1132 mg TOS/kg body weight/day. The enzyme did not exhibit, in vitro, skin, nor eye irritation potential. Acute aquatic toxicity evaluated on daphnia and algae showed absence of effect of the enzyme at the standard doses tested.Muramidase 007 was fully tolerated by broiler chickens in a 6-weeks tolerance study showing no adverse effects in any of the dietary treatments (0, 1×, 5× and 10× maximum recommended dose).In conclusion, Muramidase 007 is found to be toxicologically inert, and there are no worker's safety concerns if standard precautions are instituted and a non-dusty formulation is employed. Muramidase 007 is well tolerated by the target species (broiler chickens) and cause no harm to the environment. The beneficial safety evaluation of Muramidase 007 is in line with this type of enzyme that is found ubiquitously in nature.     

实验动物饲料硬度和粒度对饲料利用率的影响

本文简述了饲料粒度和硬度对饲料生产和对实验动物的影响.适度的粉碎粒度可以有效控制生产成本,适当的饲料粒度和硬度可以提高实验动物对饲料的利用率.     

Axo-somatic inhibition of projection neurons in the lateral nucleus of amygdala in human temporal lobe epilepsy: an ultrastructural study

Here, we report ultrastructural alterations in the synaptic circuitry of the human amygdala related to neuronal cell densities in surgical specimens of patients suffering from temporal lobe epilepsy (TLE). The neuronal cell densities quantified in the basolateral complex of amygdala were significantly reduced only in the lateral nucleus (LA) of TLE patients as compared to autopsy or non-Ammon’s horn sclerosis (AHS) controls (Nissl staining, immunostaining against the neuronal marker NeuN). For this reason, we focussed on the LA to perform a more detailed quantitative ultrastructural analysis, which revealed an inverse correlation between the number of axo-somatic inhibitory synaptic profiles at the somata of glutamic acid decarboxylase (GAD)-negative projection neurons and the extent of perisomatic fibrillary gliosis. In contrast, the density of GAD-immunoreactive interneurons positively correlated with the number of axo-somatic inhibitory synaptic profiles. The fibrillary material in perisomatic glial cell processes was preferentially labeled by the astroglial marker S100B. In addition, a qualitative study of the dendrites of GAD- and parvalbumin (PARV)-containing interneurons showed that they were often contacted by asymmetrical excitatory synapses. Our results are in line with anatomical data from rodents and cats, which show that amygdalar interneurons form axo-somatic inhibitory synapses on GAD-negative projection neurons, whereas the interneurons themselves receive excitatory input from recurrent collaterals of projection neurons and from cortico- and thalamo-amygdalar afferents. The structural reorganization patterns observed in the GABAergic circuitry are compatible with a reduced feedback or feed forward inhibition of amygdalar projection neurons in human TLE.     

Evaluation of estrogenic activity in animal diets using in vitro assay

A yeast estrogen bioassay (RIKILT REA) was in-house validated for feed on the 5 μg 17β-estradiol-equivalents per kg level according to EC Decision 2002/657/EC. All the performance characteristics met the criteria as defined in the Decision and the REA is able to detect 17β-estradiol in animal feed at a low level of 1.15–2 μg kg⁻¹. Subsequently, the developed and validated procedure was applied to determine the estrogenic activity in 24 feed samples intended for food producing animals, pets and laboratory animals. Two batches of rodent diet Murigran and one dog feed have been presented as a suspect, i.e. gave responses above the determined decision limit (CCα) and detection capability (CCβ). In assessing the performance of the estrogenic activity in these diets evaluated by comparison with the 17β-estradiol calibration curve, 17β-estradiol-equivalence levels of 7.07 μg EEQ kg⁻¹ and 9.54 μg EEQ kg⁻¹ in two batches of rodent diet and 5.3 μg EEQ kg⁻¹ in dog feed have been established. The activities observed in the rodent feed could be explained by chemical analysis, revealing high amounts of genistein, daidzein and trace amounts of zearalenone. In addition, the estrogenic activity in one of rodent feed was above the established CCα, but below the CCβ values established and all other samples showed no estrogenic activity with responses below the CCα value, which corresponds to levels below 2 μg EEQ kg⁻¹.     

Estimating meat withdrawal times in pigs exposed to melamine contaminated feed using a physiologically based pharmacokinetic model

Recently melamine was found to have contaminated the feed of multiple food production species leading to concern over the ability to establish an appropriate withdrawal interval and protect the safety of the food supply. To establish an appropriate withdrawal interval, a physiologically based pharmacokinetic (PBPK) model for melamine was developed for rats and extrapolated to pigs. The rat model underpredicted plasma concentrations, but better predicted tissue residues. Correlation values for plasma, kidney, and liver were 0.59, 0.76, and 0.73, respectively. The pig model underpredicted early plasma time points but had greater accuracy at later time points which is relevant to withdrawal times. Correlation (R²) between predicted and observed plasma values was 0.89 with a negative intercept of −0.76. The pig model estimated a withdrawal interval (based on kidney tissue residues) of 19.2 and 20.9 h for single oral exposures of 3.0 and 5.12 mg/kg of melamine, respectively. Chronic oral dosing (3.0 and 5.12 mg/kg twice daily for 7 days) yielded withdrawal intervals of 20 and 21.3 h, respectively. PBPK models, such as this one, provide evidence of the usefulness in species extrapolation over a range of dosing scenarios and can be used to protect the food supply after accidental exposure in the face of little in the target species.     

Peptides with CCK-like activity administered intracranially elicit satiety in sheep

Cholecystokinin (CCK) peptides and receptors have been shown to be present in the brain as well as in gastrointestinal organs. While functions for peripheral CCK are well recognized, those for central CCK peptides are only now being investigated. We have shown previously that CCK-octapeptide (CCK-OP) is a very potent and specific suppressor of feeding when administered in the cerebral ventricles of sheep. In the present study the objective was to determine the relative potency of several CCK analogues in inhibiting feeding when administered as 75 min continuous injections into the lateral cerebral ventricles of 2-hr fasted sheep. In comparing feeding response during CCK-OP injections to that during caerulein injections, it was found that feed intakes were similar only at an equal molar dose (0.638 pmole/min); whereas three times as much CCK-33 (1.91 pmoles/min) as CCK-OP (0.638 pmoles/min) was required to produce similar feed intakes. Both caerulein (0.638 pmoles/min) and CCK-33 (1.91 pmoles/min) caused significant decreases in feeding compared to control (sCSF). Desulfated CCK-OP had no effect on feeding at a dose (0.638 pmoles/min) that causes 80–100% decreases in feeding when the C-7 tyrosine is sulfated. Feed intake was significantly less with 2.55 pmoles/min CCK-OP than with an equal dose of desulfated CCK-OP. These results concur with those of previous studies on specific CCK receptors in the pancreas and in the brain, and therefore support the concept of specific CCK receptors in brain having a role in satiety.     

Relationship between prostagandin E1, polyphloretin phosphate and alpha and beta adrenoceptor-bound feeding loci in the hypothalamus of sheep

We have previously proposed that prostaglandins (PG) may play a modulating role on hypothalamic areas controlling feeding and energy balance. In the present experiment we have tested in the medial hypothalamus of sheep for interactions between α and β adrenoceptors, PGE₁ and a PG antagonist polyphloretin phosphate (PPP). Sheep were prepared with 6 cannula guides in the hypothalamus. In each sheep following preliminary injection with 1-norepinephrine (1-NE) and dl-isoproterenol (dl-Isop), loci were selected that showed preferentially either α or β adrenoceptor-bound feeding. In the subsequent experiment PGE₁ blocked the 1-NE (α-agonist) elicited feeding in the α-bound feeding loci but PGE₁ elicited feeding when injected into the β-bound feeding loci. The PGE₁-elicited feeding was specifically blocked by a β antagonist (LB-46). The PPP elicited feeding in both α and β-bound feeding loci but the responses were blocked only by the α antagonist (phentolamine) in the α loci and by the β antagonist (LB-46) in the β loci. These responses lend support to our previous conclusions that injection of α agonists into some and β agonists into other hypothalamic sites, but not vice versa will elicit feeding. PGE₁, injected into loci showing differences in sensitivity to adrenoceptor agonists which elicit feeding, results in increased feeding, as in this experiment, or decreased feeding as shown in a previous report. Thus, we conclude that although it is unlikely that systematically produced PG modulate hypothalamic controls on feeding and energy balance because of the dual effect on feeding, there may be an interaction of endogenous hypothalamic PG and adrenoceptors.     

Mechanisms of xenin-induced anorectic response in chicks (Gallus gallus)

We recently reported that the 25 amino acid peptide xenin caused reduced feed intake when centrally injected in chicks. The present study was designed to explore possible mechanisms of the xenin-induced anorexigenic response in chicks. In Experiments 1 and 2, chicks were implanted with cannulas and xenin injections were made directly into the ventromedialis hypothalami (VMH). Chicks responded with reduced feed intake and increased c-Fos immunoreactivity at the VMH. In Experiment 3 chicks that received co-intracerebroventricular (ICV) injection of naloxone and a dose of xenin (100 pmol), that alone does not affect feed intake, had reduced feed intake. In Experiment 4, chicks responded to ICV xenin with reduced feed- but increased exploratory-pecking. Thus, we conclude that xenin may mediate its effect directly at the VMH and that the endogenous opioid system may counter anorexigenic effects of low xenin doses in chicks. Xenin also caused increased exploration of a novel environment, an effect that may be competitive with feeding. Taken together, these results suggest that xenin regulation of chick appetite is the result of several central and behavioral mechanisms acting in synergism.     

Toxicological studies on a novel phytase expressed from synthetic genes in Aspergillus oryzae

Phytases are widely used as feed additives for monogastric animals, which cannot easily utilise the phosphorus bound in phytate (myo-inositol hexakisphosphate). The current study presents a safety evaluation of a 6-phytase produced by an Aspergillus oryzae strain expressing two synthetic genes, both mimicking a phytase gene from a Citrobacter braakii strain. Oral administration of the phytase preparation to rats at a dose level of 0.86 g total organic solids/kg body weight/day for 13 weeks did not cause any adverse effect. The phytase preparation did not exhibit irritative potential when applied locally to the eyes of rabbits or when applied to the skin using the in vitro three-dimensional epidermis model of adult human-derived epidermal keratinocytes. Furthermore, the phytase preparation was found not to represent mutagenic or clastogenic potential in the bacterial reverse mutation assay and in the in vitro micronucleus assays. Based on the toxicological data, the large safety factors calculated under common recommended dose assumptions for broiler chickens and weaned piglets, and the fact that Aspergillus oryzae is considered a safe strain lineage, it is concluded that there are no reasons for safety concerns when using this phytase as a feed additive.