The production of yeast cell wall using an agroindustrial waste influences the wall thickness and is implicated on the aflatoxin B1 adsorption process.

The objectives of this study were: to evaluate the use of dry distillery grain soluble extract - DDGse to produce yeast biomass and to obtain cell wall (CW), to use the CW as an aflatoxin B1 (AFB1) adsorbent, to study the variation in the composition and thickness of the CW under the influence of DDGse to evaluate their implication on the adsorption process using transmission electron microscopy (TEM) and fourier-transform infrared spectroscopy (FITR). The production of biomass and CW were variable. The CW thickness values showed that S. boulardii strain grown in yeast extract peptone dextrose (YPD) or DDGse medium, with no significant differences observed. The thickness of the CW for S. cerevisiae (RC012 and VM014) were increased when the cells were grown in DDGse medium, the thickness was almost double compared to the values obtained in YPD medium. The spectra IR of each CW in the two culture media shown regions corresponding to polysaccharides, proteins and lipids. Cells grown in DDGse medium adsorbed more AFB1 than those grown in YPD. The CW adsorbed more AFB1 than the same amount of whole cell. Future studies should be done to determine the type of carbohydrates and the relationship between chitin - beta glucans responsible for mycotoxin adsorption.

Genome sequence analysis of a distinctive Italian infectious bursal disease virus.

In a recent study, an emerging infectious bursal disease virus (IBDV) genotype (ITA) was detected in IBDV-live vaccinated broilers without clinical signs of infectious bursal disease (IBD). VP2 sequence analysis showed that strains of the ITA genotype clustered separately from vaccine strains and from other IBDV reference strains, either classic or very virulent. In order to obtain a more exhaustive molecular characterization of the IBDV ITA genotype and speculate on its origin, genome sequencing of the field isolate IBDV/Italy/1829/2011, previously assigned to the ITA genotype, was performed, and the sequences obtained were compared to the currently available corresponding sequences. In addition, phylogenetic and recombination analyses were performed. Interestingly, multiple amino acid (AA) sequence alignments revealed that the IBDV/Italy/1829/2011 strain shared several AA residues with very virulent IBDV strains as well as some virulence markers, especially in the VP1 protein. Nevertheless, sequence analysis demonstrated the presence of several residues typical of IBDV strains at a low degree of virulence in the IBDV/Italy/1829/2011 strain. Although homologous recombination and reassortant phenomena may occur naturally among different IBDV strains, no evidence of those events was found in the genome of the IBDV/Italy/1829/2011 strain, which was confirmed to be a genetically distinctive IBDV genotype.

A sensitive, reproducible, and economic real-time reverse transcription PCR detecting avian metapneumovirus subtypes A and B.

Use of real-time PCR is increasing in the diagnosis of infectious disease due to its sensitivity, specificity, and speed of detection. These characteristics make it particularly suited for the diagnosis of viral infections, like avian metapneumovirus (AMPV), for which effective control benefits from continuously updated knowledge of the epidemiological situation. Other real-time reverse transcription (RT)-PCRs have been published based on highly specific fluorescent dye-labeled probes, but they have high initial cost, complex validation, and a marked susceptibility to the genetic variability of their target sequence. With this in mind, we developed and validated a SYBR Green I-based quantitative RT-PCR for the detection of the two most prevalent AMPV subtypes (i.e., subtypes A and B). The assay demonstrated an analytical sensitivity comparable with that of a previously published real-time RT-PCR and the ability to detect RNA equivalent to approximately 0.5 infectious doses for both A and B subtypes. The high efficiency and linearity between viral titer and crossing point displayed for both subtypes make it suited for viral quantification. Optimization of reaction conditions and the implementation of melting curve analysis guaranteed the high specificity of the assay. The stable melting temperature difference between the two subtypes indicated the possibility of subtyping through melting temperature analysis. These characteristics make our assay a sensitive, specific, and rapid tool, enabling contemporaneous detection, quantification, and discrimination of AMPV subtype A and B.

Detection of Hepatitis E Virus (HEV) in Italian pigs displaying different pathological lesions.

In this study we investigated the HEV prevalence in Italian pigs displaying different pathological lesions, possible risk factors related to the infection, and the possible relations occurring between HEV and other concomitant pig pathogens. Genetic characterization of some of the identified strains was also performed. Detection of HEV RNA was accomplished using a nested reverse-transcription polymerase chain reaction on bile samples from 137 pigs of 2-4months of age submitted for diagnostic purposes. Forty-one of the 137 examined pigs (29.9%) tested positive for HEV RNA. Animals of 80-120days of age showed a higher prevalence of HEV infection (46.9% against 20% of younger animals). No statistically significant correlations between HEV positivity and the presence of other pathological conditions detected at necropsy, or concomitant coinfections with PCV2 and/or PRRSV were detected. All identified strains belonged to genotype 3, and were similar to other HEV subtypes 3e, 3f, 3c circulating in Europe.

Longitudinal study of Salmonella infection in Italian farrow-to-finish swine herds.

A longitudinal study of Salmonella enterica infection was carried out in five Italian farrow-to-finish swine herds previously known to be infected by Salmonella. Five litters were randomly selected from each herd and in each litter six piglets were randomly selected and individually identified. Thus, the study included 30 pigs from each farm. At weaning, individual blood samples were collected for serological examination from all selected piglets and on the same day from all sows in the farrowing unit. Piglets were bled again at approximately 60, 90, 150, 210 and 270 days of life whereas the last blood sample was collected at slaughtering. In one of the herds, in which the duration of productive cycle was about 12 months, the last blood samples were collected at 350 days of life. With the same time scheduling, five pen pooled faecal samples were collected from each herd for bacteriological examination. At slaughtering, mesenteric lymph nodes were collected from each ear-tagged pig. Sero-prevalence (cut off S/P ratio 0.25) in sows varied from 93.8% to 100%. In four herds, sero-prevalence in piglets showed a similar profile with complete decline of maternal antibodies at day 60 and clear sero-conversion between day 90 and day 150. In one herd, sero-conversion was observed earlier and 56% of piglets were positive at day 90. The peak of sero-prevalence was observed between day 210 and day 270. Sero-prevalence at slaughtering varied from 66% to 100%. Salmonella was isolated from faecal samples in four of five herds. No Salmonella was isolated from mesenteric lymph nodes at slaughter in two of the herds. Culture prevalence from mesenteric lymph nodes in the other three herds ranged from 3.3% to 30%. This longitudinal study provides original information about epidemiological dynamics of Salmonella enterica infection in Italian swine herds in consideration of the unique extended fattening period typical of the Italian production.

[Mucoregulatory therapy in secreting disease of the middle ear]. / Terapia mucoregolatrice nella patologia secernente dell'orecchio medio.

A controlled double-blind study was carried out vs placebo to evaluate the preventive and therapeutic efficacy of carboxymethylcysteine lysine salt (SCMC-Lys) in 40 non-adenoidectomised children (aged between 6-7 years old) with secretive otitis media and in 30 adult patients with acute inflammatory pathologies of the middle ear. The drug SCMC-Lys was administered to children at a dose of 750 mg/die in a linctus formula for 10 days and 2.7 g in a granular formula once a day for a maximum of 15 days if recovery had not already occurred. All subjects were assessed before and after treatment by means of an anamnestic and objective otological examination, tympanometry, nephelometric tests for IgA and mucociliary transport time. SCMC-Lys eliminated hypoacoustic symptoms in 85% of children treated with SCMC-Lys and in 35% of those treated with placebo.