Genotypic and phenotypic diversity of Alicyclobacillus acidocaldarius isolates.

UNLABELLED: The fruit juice industry recognizes Alicyclobacillus as a major quality control target micro-organism. In this study, we analysed 19 bacterial isolates to identify Alicyclobacillus species by polymerase chain reaction (PCR) and sequencing analyses. Phenotypic and genomic diversity among isolates were investigated by API 50CHB system and ERIC-PCR (enterobacterial repetitive intergenic consensus-PCR) respectively. All bacterial isolates were identified as Alicyclobacillus acidocaldarius, and almost all showed identical DNA sequences according to their 16S rRNA (rDNA) gene partial sequences. Only few carbohydrates were fermented by A. acidocaldarius isolates, and there was little variability in the biochemical profile. Genotypic fingerprinting of the A. acidocaldarius isolates showed high diversity, and clusters by ERIC-PCR were distinct to those obtained from the 16S rRNA gene phylogenetic tree. There was no correlation between phenotypic and genotypic variability in the A. acidocaldarius isolates analysed in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: Detection of Alicyclobacillus strains is imperative in fruit concentrates and juices due to the production of guaiacol. Identification of the genera originates rejection of the product by processing industry. However, not all the Alicyclobacillus species are deteriorative and hence the importance to differentiate among them. In this study, partial 16S ribosomal RNA sequence alignment allowed the differentiation of species. In addition, ERIC-PCR was introduced for the genotypic characterization of Alicyclobacillus, as an alternative for differentiation among isolates from the same species.

Dietary conjugated linoleic acid and its effect on immune response in pigs infected with the porcine reproductive and respiratory syndrome virus.

The aim of this study was to evaluate the effects of dietary conjugated linoleic acid (CLA) on immune response in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). A total of 18 pigs 4 weeks of age were allocated to 3 treatments, 6 per treatment: 0% CLA, 1% CLA, and 2% CLA. Serum IL-1ß, IL-6 and TNF-α; lymphocyte proliferation; and IL-2, IFN-γ, IL-10, IL-4 and IL-12 in PBMCs were evaluated. NF-κB, COX2, iNOS and PPAR-γ mRNA were also evaluated. No differences were observed among treatment groups in most of the in vivo cytokine profiles; only TNF-α production was increased in infected pigs in the CLA-supplemented groups. The cytokine profile in vitro was not affected by CLA supplementation. CLA decreased the proliferation of PBMCs stimulated with PRRSVs. Inflammation mediators and PPAR-γ were not affected by CLA in infected pigs. CLA did not improve the immune response of PRRSV infected pigs.

Analysis of ORF5 and full-length genome sequences of porcine reproductive and respiratory syndrome virus isolates of genotypes 1 and 2 retrieved worldwide provides evidence that recombination is a common phenomenon and may produce mosaic isolates.

UNLABELLED: Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) isolates by examining previously published data sets of ORF5 sequences (genotypes 1 and 2) obtained worldwide. We then examined full-length genome sequences in order to determine potential recombination breakpoints along the viral genome. For ORF5, 11 sets of genotype 1 sequences from different geographical areas, including 2 Asian, 1 American, and 7 European regions, and three sets of genotype 2, including sets from China, Mexico, and the United States, were analyzed separately. Potential recombination breakpoints were detected in 10/11 genotype 1 sets, including 9 cases in which the clustering of at least one isolate was different before and after the breakpoints. In genotype 2, potential breakpoints and different tree clustering of at least one strain before and after the breakpoint were observed in 2 out of 3 sets. The results indicated that most of the ORF5 data sets contained at least one recombinant sequence. When the full-length genome sequences were examined, both genotype 1 and 2 sets presented breakpoints (10 and 9, respectively), resulting in significantly different topologies before and after the breakpoints. Mosaic genomes were detected in genotype 1 sequences. These results may have significant implications for the understanding of the molecular epidemiology of PRRSV. IMPORTANCE: PRRSV is one of the most important viruses affecting swine production worldwide, causing big economic losses and sanitary problems. One of the key questions on PRRSV arises from its genetic diversity, which is thought to have a direct impact on immunobiology, epidemiology, diagnosis, and vaccine efficacy. One of the causes of this genetic diversity is recombination among strains. This study provides evidence that recombinant PRRSV isolates are common in most of the countries with significant swine production, especially PRRSV genotype 1. This observation has implications in the proper characterization of PRRSV strains, in the future development of phylogenetic studies, and in the development of new PRRSV control strategies. Moreover, the present paper emphasizes the need for a deeper understanding of the mechanisms and circumstances involved in the generation of genetic diversity of PRRSV.

Naturally co-infected boars with both porcine reproductive and respiratory syndrome virus and porcine circovirus type 2.

In this study, the humoral response against porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2), the presence of the virus in semen and serum and the genetic characteristics of the virus detected in 15 boars from a commercial farm were analysed. The results showed that 53% of the boars presented anti-PRRSV antibodies and 100% presented anti-PCV2 antibodies. Porcine reproductive and respiratory syndrome virus was detected in 43% of the boars and 73% were positive to PCV2. The complete ORF5 gene of PRRSV of 14 samples and a fragment of the ORF2 gene of PCV2 of 22 samples were sequenced. Porcine reproductive and respiratory syndrome virus analysis revealed <92% identity in viruses from semen and serum of two boars, whereas in the rest of the boars the identity was >97.5%. As for PCV2, two boars presented an identity <95% in serum and semen and the rest had an identity >96%. The results showed that PRRSV- and PCV2-naturally infected boars can be found, and at least two different strains of viruses from semen and serum can be detected.