Better understanding of the molecular changes associated with disease is essential for identifying new routes to improved therapeutics and diagnostic tests. The aim of this study was to investigate the dynamic changes in the metabolic profile of mouse sera during T. gondii infection. We carried out untargeted metabolomic analysis of sera collected from female BALB/c mice experimentally infected with the T. gondii Pru strain (Genotype II). Serum samples were collected at 7, 14 and 21 day post infection (DPI) from infected and control mice and were subjected to liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS)-based global metabolomics analysis. Multivariate statistical analysis identified 79 differentially expressed metabolites in ESI+ mode and 74 in ESI− mode in sera of T. gondii-infected mice compared to the control mice. Further principal component analysis (PCA) and partial least squares-discrimination analysis (PLS-DA) identified 19 dysregulated metabolites (5 in ESI+ mode and 14 in ESI− mode) related to the metabolism of amino acids and energy metabolism. The potential utility of these metabolites as diagnostic biomarkers was validated through receiver operating characteristic (ROC) curve analysis. These findings provide putative metabolite biomarkers for future study and allow for hypothesis generation about the pathophysiology of toxoplasmosis.

Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical properties of nucleic acids. Furthermore, this protocol showed that DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl) during lysis step, and this environment is improper for RNase to have DNA liberated of RNA and even for DNase to degrade the DNA. Therefore, the complete removal of RNA under RNase influence is achieved when RNase is added after DNA extraction, which gives optimal quality with any protocols. Similarly, DNA contamination in an isolated RNA is degraded by DNase to obtain high-quality RNA. Our protocol is the protocol of choice in terms of simplicity, recovery time, environmental safety, amount, purity, PCR and RT-PCR applicability.

Diarrhea remains one of the leading causes of morbidity and mortality globally, with enterotoxigenic Escherichia coli (ETEC) constituting a major causative pathogen. The development of alternative treatments for diarrhea that do not involve chemotherapeutic drugs or result in antibiotic resistance is critical. Considering that lysozyme is a naturally occurring antimicrobial peptide, in a previous study we developed a transgenic pig line that expresses recombinant human lysozyme (hLZ) in its milk. In the present study, we examined the protective effects of the consumption of this milk against ETEC infection in neonatal piglets. We found that consuming hLZ milk facilitated faster recovery from infection and decreased mortality and morbidity following an ETEC oral inoculation or infection acquired by contact-exposure. The protective effect of hLZ was associated with the enrichment of intestinal bacteria that improve gut health, such as Lactobacillus, and the enhancement of the mucosal IgA response to the ETEC-induced diarrhea. Our study revealed potential protective mechanisms underlying the antimicrobial activity of human lysozyme, validating the use of lysozyme as an effective preventive measure for diarrhea.

Understanding the occurrence and distribution of various Eimeria species in broiler farms is necessary to develop effective coccidiosis vaccines. In the current study, fecal samples were collected from broilers with subclinical signs at fifty small-scale farms in the Shandong province in eastern China. Oocysts purified from fecal samples were examined for morphology. The Eimeria genomic DNA extracted from each sample was subjected to PCR amplification with species-specific primers for the internal transcribed spacer sequence or the small RNA subunit sequence of each of the seven species of Eimeria found in chickens. The results showed that more than one Eimeria species existed in most fecal samples, and the infection rate of identified Eimeria spp. in these farms was 90%, 88%, 72%, 68%, 60%, 26%, and 8% for E. tenella, E. praecox, E. acervulina, E. maxima, E. mitis, E. necatrix, and E. brunetti, respectively. This indicates that E. tenella, E. praecox, E. acervulina, E. maxima, and E. mitis are t... Resumen Nota de Investigación—Prevalencia de especies de Eimeria en pollos de engorde con signos subclínicos en 50 granjas. Es necesario entender la presentación y distribución de varias especies de Eimeria en granjas de pollo de engorde para desarrollar programas efectivos de vacunación. En el presente estudio, se recolectaron muestras fecales de pollos de engorde con signos subclínicos en 50 granjas pequeñas ubicadas en la provincia de Shandong en el este de China. Los oocystos purificados de las muestras fecales fueron examinados en su morfología. El ADN genómico de Eimeria extraído de cada muestra fue sometido a amplificación mediante la reacción en cadena de la polimerasa (de las siglas en Inglés PCR) utilizando iniciadores específicos de especie. Dichos iniciadores estaban dirigidos para amplificar la secuencia espaciadora transcripta interna o la secuencia de la subunidad menor de ARN de cada una de las siete especies de Eimeria que afectan a los pollos. Los resultados demostraron que más de una es...

The “self-cleaving” 2A sequence of picornavirus, which mediates ribosome-skipping events, enables the generation of two or more separate peptide products from one mRNA containing one or more “self-cleaving” 2A sequences. In this study, we introduced a single 2A sequence of porcine teschovirus-1 (P2A) linked to two fluorescent protein genes, the enhanced yellow fluorescent protein (EYFP) gene and the red fluorescent protein (RFP) gene, in a single cassette into transgenic Eimeria tenella (EtER). As expected, we obtained two separated protein molecules rather than a fused protein, although the two molecules were translated from the same mRNA carrying a single “self-cleaving” 2A sequence. Importantly, RFP led by a secretion signal was secreted into parasitophorous vacuoles, while EYFP localized mainly to the nucleus of EtER. Our results demonstrate that the “self-cleaving” 2A sequence actively mediated cleavage of polyproteins in the apicomplexan parasite E. tenella.