Association between canine leishmaniosis and Ehrlichia canis co-infection: a prospective case-control study

Abstract Background In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. Methods We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. Results From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or “Candidatus Mycoplasma haematoparvum” DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5–106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to be co-infected with M. haemocanis. Conclusions Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR

Characterization of the effects of Atosiban on uterine electromyograms recorded in women with threatened preterm labor

[EN] Although research studies using electrohysterography on women without tocolytic therapy have shown its potential for preterm birth diagnosis, tocolytics are usually administered in emergency rooms at the first sign of threatened preterm labor (TPL). Information on the uterine response during tocolytic treatment could prove useful for the development of tools able to predict true preterm deliveries under normal clinical conditions. The aim of this study was thus to analyze the effects of Atosiban on Electrohysterogram (EHG) parameters and to compare its effects on women who delivered preterm (WDP) and at term (WDT). Electrohysterograms recorded in different Atosiban therapy stages (before, during and after drug administration) on 40 WDT and 27 WDP were analyzed by computing linear, and non-linear EHG parameters. Results reveal that Atosiban does not greatly affect the EHG signal amplitude, but does modify its spectral content and reduces the energy associated with the fast wave high component in both WDP and WDT, with a faster response in the latter. EHG signal complexity remained constant in WDT, while it increased in WDP until it reached similar values to WDT during Atosiban treatment. The spectral and complexity parameters were able to separate (p < 0.05) WDT and WDP prior to and during tocolytic treatment and before and after treatment, respectively. The results pave the way for developing better and more reliable medical decision support systems based on EHG for preterm delivery prediction in TPL women in clinical scenarios.This work received financial support from the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (DPI2015-68397-R), VLC/Campus (UPV-FE-2018-B03) and by Conselleria de Educación, Investigación, Cultura y Deporte, Generalitat Valenciana (GV/2018/104).Mas-Cabo, J.; Prats-Boluda, G.; Ye Lin, Y.; Alberola Rubio, J.; Perales, A.; Garcia-Casado, J. (2019). Characterization of the effects of Atosiban on uterine electromyograms recorded in women with threatened preterm labor. Biomedical Signal Processing and Control. 52:198-205. https://doi.org/10.1016/j.bspc.2019.04.001S1982055

<it>Toxoplasma gondii</it> sexual cross in a single naturally infected feline host: Generation of highly mouse-virulent and avirulent clones, genotypically different from clonal types I, II and III

<p>Abstract</p> <p>Tachyzoite clones obtained from a single <it>Toxoplasma gondii</it> oocyst field sample were genotyped and characterized regarding mouse virulence. PCR-RFLP genotyping of tachyzoites initially isolated from interferon-γ-knockout (GKO) mice, BALB/c mice and VERO cell culture using the nine independent, unlinked genetic markers nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico revealed mixed <it>T. gondii</it> infections showing combinations of type II and type III alleles at different loci. Forty-five individual clones were obtained from all mixed <it>T. gondii</it> tachyzoite cell cultures by limiting dilution. Sixteen <it>T. gondii</it> clones showed type III alleles at all loci and 29 clones displayed a combination of type II and type III alleles at different loci. Five clone groups were identified in total, four of which include <it>T. gondii</it> clones that showed a non-canonical allele pattern and have never been described in natural infections before. All tested clones, except two, were highly virulent in BALB/c mice. The isolation of different non-canonical <it>T. gondii</it> clones originating from an oocyst sample of a single naturally infected cat demonstrate that sexual recombination as well as re-assortment of chromosomes via a sexual cross of <it>T. gondii</it> occur under natural conditions and result in the emergence of clones with increased virulence in mice.</p

Lungworm infections (Angiostrongylus vasorum, Crenosoma vulpis, Aelurostrongylus abstrusus) in dogs and cats in Germany and Denmark in 2003-2007

Faecal samples of 4151 dogs from Denmark, 958 clogs from Germany and 231 cats from Germany with clinical signs were examined for lungworm larvae using the Baermann funnel technique between 2003 and 2007. In total, 3.6% of Danish and German dogs shed lungworm larvae. In Denmark, patent infections of clogs with Angiostrongylus vasorum were more prevalent (2.2%) than those with Crenosoma vulpis (1.4%). In Denmark, the majority of A. vasorum- (98%) and C. vulpis-infected (80%) clogs originated from Northern Zealand. The frequency of A. vasorum and C. vulpis infections in Danish dogs obviously decreased from 2003 to 2006. In Germany, canine faecal samples were found more frequently positive for C. vulpis than for A. vasorum larvae (2.4% and 1.2%, respectively). Lungworm-infected dogs originated mainly from Southern and western Germany. Larvae of Aelurostrongylus abstrusus were detected in 5.6% of cats from Germany. Overall, a distinct seasonal pattern in the detection of infected dogs was apparent for A. vasorum in Denmark and C. vulpis in Germany. The relatively high number of lungworm-infected dogs and cats indicate that these parasitic diseases should be considered in differential diagnosis of cases of treatment-resistant respiratory/cardiopulmonary distress. (C) 2008 Elsevier B.V. All rights reserved

Genotyping of European Toxoplasma gondii strains by a new high-resolution next-generation sequencing-based method.

PURPOSE A new high-resolution next-generation sequencing (NGS)-based method was established to type closely related European type II Toxoplasma gondii strains. METHODS T. gondii field isolates were collected from different parts of Europe and assessed by whole genome sequencing (WGS). In comparison to ME49 (a type II reference strain), highly polymorphic regions (HPRs) were identified, showing a considerable number of single nucleotide polymorphisms (SNPs). After confirmation by Sanger sequencing, 18 HPRs were used to design a primer panel for multiplex PCR to establish a multilocus Ion AmpliSeq typing method. Toxoplasma gondii isolates and T. gondii present in clinical samples were typed with the new method. The sensitivity of the method was tested with serially diluted reference DNA samples. RESULTS Among type II specimens, the method could differentiate the same number of haplotypes as the reference standard, microsatellite (MS) typing. Passages of the same isolates and specimens originating from abortion outbreaks were identified as identical. In addition, seven different genotypes, two atypical and two recombinant specimens were clearly distinguished from each other by the method. Furthermore, almost all SNPs detected by the Ion AmpliSeq method corresponded to those expected based on WGS. By testing serially diluted DNA samples, the method exhibited a similar analytical sensitivity as MS typing. CONCLUSION The new method can distinguish different T. gondii genotypes and detect intra-genotype variability among European type II T. gondii strains. Furthermore, with WGS data additional target regions can be added to the method to potentially increase typing resolution