Peanut allergen Ara h 1 interacts with proanthocyanidins into higher molecular weight complexes

Mildly extracted peanut allergen Ara h 1 was previously reported to occur as an oligomeric complex. In this paper we describe how the protein in this oligomeric complex interacts noncovalently with phenolic compounds of the proanthocyanidin type. These interactions are being disrupted during anion exchange chromatography, resulting in the dissociation of the oligomeric Ara h 1 complex into protein trimers. By use of the known three-dimensional structure of beta-conglycinin, a soy protein homologous to Ara h 1, proline-rich regions were observed in silico on both faces of its trimeric structure, which are conserved in Ara h 1. These proline-rich regions could explain the binding of proanthocyanidins to Ara h 1 and the formation of multiple Ara h 1 trimer complexes. This was supported by the observation that the addition of peanut proanthocyanidins to trimeric Ara h 1 and to beta-conglycinin resulted in the formation of soluble oligomeric protein complexes. The structurally related legumin proteins do not contain such proline-rich regions on both sides of the protein, and proanthocyanidins were shown to have a lower affinity for legumin proteins from peanuts and soybeans (peanut allergen Ara h 3 and soy glycinin, respectively). Ara h 1 present as the oligomeric complex is assumed to be the representative form of the allergen in which it is consumed by humans.     

Phylogenetic analysis of the SSU rRNA gene from the piscine diplomonad Spironucleus torosus (Diplomonadida: Hexamitinae)

Previous studies have recorded Spironucleus torosus Poynton et Morrison, 1990 from several species of gadoid fishes, including the only freshwater gadoid, the burbot Lota lota (L.). Two morphologically different isolates of S. torosus have been described (elongate and pyriform). Both have been found in saltwater, while only the elongate has been found in freshwater. To address the conspecificity of the two morphs of S. torosus, and to identify the source of S. torosus in burbot in Norway, we have sequenced the small subunit ribosomal RNA (SSU rRNA) gene from 43 isolates of S. torosus from six species of gadoid fishes sampled at 15 localities in Norway, Sweden and the Baltic Sea. Phylogenetic analyses of the SSU rRNA gene sequence data recovered two major clades, one containing mainly isolates from burbot, while the other contained isolates from marine gadoid fishes only. The genetic distance (based on 25 nucleotide substitutions in 789 base pairs) separating the two assemblages was not large enough to consider the two groups separate species. Spironucleus torosus isolated from burbot displayed limited genetic variation in the small subunit ribosomal RNA (SSU rRNA) gene along the post-Pleistocene migration route of its host. The present study is the first report of S. torosus in tusk Brosme brosme (Ascanius), whiting Merlangius merlangus (L.), and fourbeard rockling Enchelyopus cimbrius (L.).     

Litter production and nutrient deposition from native woody species in the Brazilian semi-arid region

Agroforestry Systems - The use of native trees in agroecosystems is a promising way to increase litter deposition and nutrient cycling and foster the recovery of degraded areas, especially in...     

Radiographic evaluation of positional atelectasis in sedated dogs breathing room air versus 100% oxygen

This study documents the degree of positional atelectasis in sedated dogs receiving 100% oxygen (O₂) versus room air. Initial lateral recumbency was determined by an orthopedic study and initial treatment (O₂ or room air) was randomized. Each dog was maintained in lateral recumbency for 15 min, at which time ventrodorsal (VD) and opposite lateral thoracic radiographs were obtained. Each dog was then maintained in the opposite lateral recumbency and received the other treatment for 15 min, followed by a VD and opposite lateral radiograph. Radiographs were scored for severity of pulmonary pattern and mediastinal shift by 3 radiologists. Dogs breathing O₂ had significantly higher scores than dogs breathing room air. If radiographically detectable dependent atelectasis is present, repeat thoracic images following manual positive ventilation and/or position change to the opposite lateral recumbency should be made to rule out the effect of O₂ positional atelectasis and avoid misdiagnosis.     

Herd diagnosis of low pathogen diarrhoea in growing pigs – a pilot study

Background

The major indication for antibiotic use in Danish pigs is treatment of intestinal diseases post weaning. Clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. In some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). Termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce the consumption of antibiotics in the pig industry. The objective of the present pilot study was to suggest criteria for herd diagnosis of low pathogen diarrhoea in growing pigs.Data previously collected from 20 Danish herds were used to create a case series of clinical diarrhoea outbreaks normally subjected to antibiotic treatment. In the present study, these diarrhoea outbreaks were classified as low pathogen (<15% of the pigs having bacterial intestinal disease) (n =5 outbreaks) or high pathogen (≥15% of the pigs having bacterial intestinal disease) (n =15 outbreaks). Based on the case series, different diagnostic procedures were explored, and criteria for herd diagnosis of low pathogen diarrhoea were suggested. The effect of sampling variation was explored by simulation.

Results

The diagnostic procedure with the highest combined herd-level sensitivity and specificity was qPCR testing of a pooled sample containing 20 randomly selected faecal samples. The criteria for a positive test result (high pathogen diarrhoea outbreak) were an average of 1.5 diarrhoeic faecal pools on the floor of each pen in the room under investigation and a pathogenic bacterial load ≥35,000 per gram in the faecal pool tested by qPCR. The bacterial load was the sum of Lawsonia intracellularis, Brachyspira pilosicoli and Escherichia coli F4 and F18 bacteria per gram faeces. The herd-diagnostic performance was (herd-level) diagnostic sensitivity =0.99, diagnostic specificity =0.80, positive predictive value =0.94 and negative predictive value =0.96.

Conclusions

The pilot study suggests criteria for herd diagnosis of low pathogen diarrhoea in growing pigs. The suggested criteria should now be evaluated, and the effect of terminating antibiotic batch medication in herds identified as suffering from low pathogen diarrhoea should be explored.

Electronic supplementary material

The online version of this article (doi:10.1186/2046-0481-67-24) contains supplementary material, which is available to authorized users.     

Field evaluation of live chilling with CO2 on stunning Atlantic salmon (Salmo salar) and the subsequent effect on quality

Atlantic salmon were slaughtered in three ways on a commercial slaughter line: (1) killed by a percussive stun after crowding; (2) killed by percussive stun after crowding, pumping and live chilling; (3) killed by exsanguination after crowding, pumping and live chilling. The live‐chilled fish were exposed to seawater (2°C) saturated with carbon dioxide (pH 5.5–5.7) for 40 min. The fish were calm after live chilling, but not unconscious, as eye rolling was observed in all individuals. Subsequent exsanguination of the unstunned fish resulted in death. Both rapid live chilling and the subsequent exsanguination appeared stressful to the fish, as a large and rapid pH drop coupled with earlier onset of rigor mortis, indicative of high muscle activity during the process were observed. The muscle core temperature during ice storage showed that live chilling only has an effect on carcass temperature during the first 6 h post mortem. After 6 h, no significant differences in temperature were detected between live‐chilled and traditionally ice‐chilled fish. We conclude that commercial use of live chilling in combination with high levels of CO₂ does not stun Atlantic salmon. Live chilling followed by exsanguination of the unstunned fish appears to be highly stressful and should be avoided.     

The effect of slaughtering procedures on blood spotting in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

Rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) were submitted for various slaughter and bleeding procedures to see what effect this would have on blood drainage of the muscles. Results show that the bleeding method is of less importance, while it is the timing that is important. No significant difference in bloodspotting was observed between fish that were bled live by a gill cut or percussive killed and bled by gutting. Most of the drainage of blood in the fish muscle seems to occur within the first hours postmortem, so rigor mortis is of little importance. The visual appearance of the fillet was influenced by number and size of the bloodstains. Colour measurements with Hunter L*, a*, b* did not reveal this. We conclude that a gill cut is not necessarily to obtain bleeding, so the industry can omit this phase and go directly to gutting.     

Steroidogenesis during induced oocyte maturation and ovulation in the African catfish,Clarias gariepinus

Changes in ovarian steroidogenesis accompanying oocyte maturation and ovulation were studied in the African catfish,Clarias gariepinus. Laboratory-reared females with postvitellogenic ovaries were treated with pimozide and LHRH-analogue. The plasma gonadotropin levels were determined by means of a homologous radioimmunoassay, the condition of the ovaries was studied by histological examination of the follicles, and the steroidogenetic capacity of the ovaries was analyzed byin vitro incubation of tissue fragments for 3 h with [³H]-pregnenolone and [³H]androstenedione as precursors. Data were collected at regular intervals between 0 and 16 h after pimozide-LHRH analogue administration.Until 4 h after the beginning of the experiments the plasma gonadotropin levels did not rise above 25 ng/ml, the ovaries remained in the stage of postvitellogenesis, and testosterone was the main end product of steroidogenesis. Four hours later the gonadotropin concentration in the blood had risen to more than 150 ng/ml, and the ovaries had entered the stage of germinal vesicle migration. At the same time steroidogenesis shifted towards the production of 17,20-dihydroxy-4-pregnen-3-one, 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,6,17,20-tetrol. During the subsequent stage of germinal vesicle breakdown the plasma gonadotropin level remained high, and the synthesis of the C₂₁-steroids showed a further increase. Simultaneously, the production of some C₁₉-steroid glucuronides was enhanced. The preovulation and especially the postovulation stages were accompanied by a gradual decrease in steroidogenic capacity of the ovaries, even though the plasma gonadotropin level remained high. It is concluded that the prematuration surge of gonadotropin influences the activity of enzymes involved in steroidogenesis, leading to a reduced C_17,20-lyase and to an augmented activity of the enzymes 20-hydroxysteroid dehydrogenase (HSD), 5-reductase, 3-HSD, 6-hydroxylase and UDP-glucuronosyltransferase. During ovulation the activity of all steroidogenic enzymes, including such key enzymes as 3-HSD and 17-hydroxylase, gradually decreases.Not only 17,20-dihydroxy-4-pregnen-3-one, but also the 5-reduced pregnanes may be involved in inducing oocyte maturation and/or ovulation. The very polar triol and tetrol products may function, together with the steroid glucuronides as sex pheromones.A preliminary account of these results was presented at the XIII Conference of European Comparative Endocrinologists, Belgrade, September 7–12, 1986