M-phase promoting factor (MPF) and mitogen activated protein kinases (MAPK) activities of domestic cat oocytes matured <i>in vitro</i> and <i>in vivo</i>

This work was undertaken in order to examine M-phase promoting factor (MPF) and mitogen-activated protein kinases (MAPK) activities during meiotic progression of cat oocytes cultured in two different media for two different incubation times and preovulatory cat oocytes that reached MII in vivo. Oocytes recovered from ovaries of ovariectomized cats were cultured either in TCM 199 or SOF for 24 h and 40 h. In vivo matured oocytes were recovered by follicular aspiration from ovaries of domestic cats ovariectomized 24 h to 26 h after hormonal treatment. Results showed that the kinetic of MPF and MAPK activity was similar during meiotic progression of cat oocytes matured in TCM 199 and SOF. After 24 h of incubation, MII oocytes had significantly (p &lt; 0.001) higher MPF and MAPK levels than MII oocytes cultured for 40 h in both culture media. MPF and MAPK activity was significantly (p &lt; 0.01) lower in the oocytes matured in vitro than in those matured in vivo. This study provides evidence that the two different maturation media did not determine differences in MPF and MAPK fluctuations and levels during meiotic progression of cat oocytes and that the time of maturation influenced the level of the two kinases. Moreover, it shows that MPF and MPK activity is higher in in vivo matured oocytes than in in vitro matured oocytes, suggesting a possible incomplete cytoplasmic maturation after culture

1h nmr brain metabonomics of scrapie exposed sheep

While neurochemical metabolite modifications, determined by different techniques, have been diffusely reported in human and mice brains affected by transmissible spongiform encephalopathies (TSEs), this aspect has been little studied in the natural animal hosts with the same pathological conditions so far

LTalpha and LTbeta gene expression in organs of sheep showing different lymphoproliferative changes induced by maedi-visna virus

In lung and mammary gland of sheep, Maedi-Visna virus (MVV) causes lymphoproliferative inflammation often with follicular structures (lymphofollicular inflammation). The aim of this work was to define whether Limphotoxin α and β (LTα, LTβ) play a role in the formation of these peculiar lesions in sheep experimentally infected with MVV

Additional polymorphisms of the <i>PRNP</i> gene significantly decrease the susceptibility to scrapie of ARQ/ARQ sheep

The aim of this work was to investigate the risk of scrapie of the ARQ/ARQ genotype carrying at least one point mutation at codons 112, 137, 141, 142, 154 and 176 in comparison with the ARQ/ARQ without any point mutations

PrP^Sc deposition in mammary gland of sheep experimentally coinfected with scrapie and Maedi-Visna virus

Scrapie, a fatal neurodegenerative disorder of sheep, is characterized by deposition of an abnormal isoform of prion protein (PrPSc) in the central nervous system (CNS) and within the lymphoreticular system (LRS). Recent studies in mice transgenically engineered to develop organ specific inflammation demonstrated the cooccurrence of PrPSc in the inflamed organs (kidney, pancreas and liver)

Intraepithelial and Interstitial Deposition of Pathological Prion Protein in Kidneys of Scrapie-Affected Sheep

Prions have been documented in extra-neuronal and extra-lymphatic tissues of humans and various ruminants affected by Transmissible Spongiform Encephalopathy (TSE). The presence of prion infectivity detected in cervid and ovine blood tempted us to reason that kidney, the organ filtrating blood derived proteins, may accumulate disease associated PrPSc. We collected and screened kidneys of experimentally, naturally scrapie-affected and control sheep for renal deposition of PrPSc from distinct, geographically separated flocks. By performing Western blot, PET blot analysis and immunohistochemistry we found intraepithelial (cortex, medulla and papilla) and occasional interstitial (papilla) deposition of PrPSc in kidneys of scrapie-affected sheep. Interestingly, glomerula lacked detectable signals indicative of PrPSc. PrPSc was also detected in kidneys of subclinical sheep, but to significantly lower degree. Depending on the stage of the disease the incidence of PrPSc in kidney varied from approximately 27% (subclinical) to 73.6% (clinical) in naturally scrapie-affected sheep. Kidneys from flocks without scrapie outbreak were devoid of PrPSc. Here we demonstrate unexpectedly frequent deposition of high levels of PrPSc in ovine kidneys of various flocks. Renal deposition of PrPSc is likely to be a pre-requisite enabling prionuria, a possible co-factor of horizontal prion-transmission in sheep

Comparison and evaluation of analytic and diagnostic performances of four commercial kits for the detection of antibodies against Echinococcus granulosus and multilocularis in human sera.

Cystic echinococcosis (CE) is a disease caused by Echinococcus granulosus sensu lato (s.l.), an ubiquitous worldwide zoonotic agent affecting humans and animals. Diagnosis of CE in humans is usually performed by imagine techniques along with immunoassays. The aim of our study was to evaluate and compare four commercial diagnostic kits, based on the detection of IgG antibodies against E. granulosus and E. multilocularis. The study was performed on a total of 259 sera: the positive (n = 74) and the negative (n = 185) group. The following analytic and diagnostic performances of the four kits were evaluated: operator skills, specificity, sensitivity, repeatability, reproducibility, accuracy, positive and negative predictive values. Based on the parameters evaluated, all four tests demonstrated excellent quality and proved to be reliable diagnostic tools to support the clinical evaluation of human patients suspected of having CE. The four commercial assays, in our hands, presented altogether, a range of performances from good to excellent, being immunoblotting (IB) the most reliable, used as gold standard, followed by the immunochromatographic test (ICT) and finally the two enzyme linked immunosorbent assay (ELISAs)

Identification of <i>Echinococcus granulosus</i> Genotypes G1 and G3 by SNPs Genotyping Assays

Echinococcus granulosus sensu lato (s.l.) is the causative agent of cystic echinococcosis in animals and humans. Different E. granulosuss.l. genotypes exhibit great diversity in their life cycle, host selectivity and pathogenicity. For this reason, the study of genetic variation within Echinococcus species is of importance for their epidemiological implication. We employed two SNP genotyping technologies to distinguish G1 and G3 E. granulosus sensu stricto (s.s.). genotypes. The genotypes of DNA samples (n = 28) extracted from hydatid cysts of different animal species were identified by amplification and sequencing of a fragment of the mitochondrial nad5 gene. Two SYBR green and three TaqMan real time PCR assays were developed for targeting of three nad5 informative positions (SNP758, 1123, and 1380) known to be able to discriminate G1 from G3. Genotyping by SYBR Green PCR based on cycle threshold (Ct) with melting temperature (Tm) analysis and performed on SNP1123 and SNP1380 failed to identify one DNA sample. TaqMan assays for SNP758, 1123 and 1380 effectively confirmed genotype identification obtained by Sanger sequencing. Our results demonstrated that the combination of the three Taqman assays developed in this study represents a valuable and cost effective tool alternative to DNA sequencing for E. granulosus s.s. genotyping

Environmental Influence on the Occurrence of Multi-Organ Cystic Echinococcosis Infection in a Patient from Sardinia, Italy

An uncommon clinical case of an adult woman who was referred to the hospital with severe symptoms attributable to cystic echinococcosis (CE) is described in this report. According to a questionnaire, the subject was exposed to a high risk of infection since she was employed on a farm about 20 years before diagnosis. She lived close to several animal species and handled vegetables in inadequate hygienic conditions. Medical and laboratory investigations confirmed the presence of massive echinococcal cystic lesions in each lung and in the liver. Given the peculiarity of the case, pharmacological and surgical treatments were the only conceivable option. The association of pharmacological treatment, surgery, and interventional radiology procedure represented a reliable and effective way to handle a complex case of human hydatidosis. A multi-disciplinary approach was mandatory, resulting in a clear and conclusive diagnosis of CE caused by the zoonotic parasite E. granulosus sensu stricto of the G1 genotype

Clinical and cytogenetic studies in intersex ewes

Nine Sarda x Lacaune ewes with intersexual characteristics and an infertility condition at the reproductive anamnesis were analysed. In order to make a diagnosis, we have evaluated their behaviour and performed clinical and laparoscopic examination of the reproductive tract, as well as cytogenetic analysis. The ewes showed basically a female phenotype but a clinical examination revealed a different degree of masculinization in the morphology of external genital organs. A shorter vagina was observed in female-like ewes and a hypertrophic clitoris in male-like ewes. Laparoscopic analysis evidenced the presence of testis in seven individuals and, for two of them, the gonadal position was subcutaneous. Different male characteristics in the nine subjects, were also observed in their behaviour with a different degree of masculinization. Their blood samples were used for determining the percentage of male cells on lymphocytes chromosome spreads by using the C-banding technique. The haematopoietic chimeras (XX/ XY) found in the lymphocytes confirmed the diagnosis of freemartinism for seven out of the nine subjects