evaluation of three commercial rapid kits to detect cryptosporidium parvum in diarrhoeic calf stool

The aim of this study was to evaluate three commercially available rapid immunochromatographic tests for detection of Cryptosporidium parvum antigens in faeces of naturally infected neonatal diarrhoeic calves. FASTest® CRYPTO strip, FASTest® CRYPTO-GIARDIA Strip and TETRASTRIPS® were compared for their sensitivity, specificity, positive predictive value and negative predictive value using a cumulative positivity as gold standard. In addition, the agreement between each test and the gold standard was evaluated by Cohen's Kappa (k) value. The highest infection rate was observed by FASTest® CRYPTO-GIARDIA Strip (65.15%), followed by FASTest® CRYPTO strip (63.64%) and TETRASTRIPS® (56.06%,). A very good diagnostic performance of all the three tests was observed. FASTest® CRYPTO strip (k = 0.935) and FASTest® CRYPTO-GIARDIA Strip (k = 0.968) had the highest sensitivity (100%) while TETRASTRIPS® (k = 0.875) had the highest specificity (100%). Eimeria spp oocysts were present in six samples but cross-reaction with this protozoan was not observed. These assays were not time-consuming and very easy to perform and to read. Based on our results, we recommend the use of FASTest® CRYPTO strip, FASTest® CRYPTO-GIARDIA Strip or/and TETRASTRIPS® for detection of C. parvum antigens in faeces of neonatal diarrhoeic calves

Evaluation of three commercial rapid kits to detect Cryptosporidium parvum in diarrheic calf stool

The aim of this study was to evaluate three commercially available rapid immunochromato-graphic tests for detection of Cryptosporidium parvum antigens in faeces of naturally infected neonatal diarrhoeic calves. FASTest (R) CRYPTO strip, FASTest (R) CRYPTO-GIARDIA Strip and TETRASTRIPS (R) were compared for their sensitivity, specificity, positive predictive value and negative predictive value using a cumulative positivity as gold standard. In addition, the agreement between each test and the gold standard was evaluated by Cohen's Kappa (k) value. The highest infection rate was observed by FASTest (R) CRYPTO GIARDIA Strip (65.15%), followed by FASTest (R) CRYPTO strip (63.64%) and TETRASTRIPS (R) (56.06%,). A very good diagnostic performance of all the three tests was observed. FASTest (R) CRYPTO strip (k= 0.935) and FASTest (R) CRYPTO-GIARDIA Strip (k= 0.968) had the highest sensitivity (100%) while TETRASTRIPS (R) (k= 0.875) had the highest specificity (100%). Eimeria spp oocysts were present in six samples but cross-reaction with this protozoan was not observed. These assays were not time-consuming and very easy to perform and to read. Based on our results, we recommend the use of FA

Distribution of different HBV DNA forms in plasma and peripheral blood mononuclear cells (PBMCs) of chronically infected patients with low or undetectable HBV plasma viremia

Few studies have documented hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMCs). We developed real-time PCR methods for differential amplification of covalently closed circular (cccDNA) and total HBV DNA (tDNA). The different distribution of cccDNA and tDNA in plasma and PBMCs was evaluated in 37 patients with low or undetectable viremia. Plasma tDNA measured by the Abbott reference system and the in-house assay correlated well (Spearman rho = 0.804; P<0.0001). tDNA was detected in four PBMC samples, all from patients with detectable plasma viremia (range 633-6,406 IU/ml), cccDNA was not detected in any sample. The reasons for apparently discrepant results need further investigation but possibly include the high diversification of HBV status and plasma viremia levels

Case report: Persistent strongyloidiasis complicated by recurrent meningitis in an HTLV seropositive Peruvian migrant resettled in Italy

We describe a case of persistent strongyloidiasis complicated by recurrent meningitis, in a human T cell lymphotropic virus type 1 (HTLV-1) seropositive Peruvian migrant adult resettled in Italy. He was admitted with signs and symptoms of acute bacterial meningitis, reporting four other meningitis episodes in the past 6 years, with an etiological diagnosis of Escherichia coli and Enterococcus faecium in two cases. He had been previously treated with several antihelmintic regimens not including ivermectin, without eradication of strongyloidiasis, and he had never been tested for HTLV before. During the described episode, the patient was treated for meningitis with broad-spectrum antibiotic therapy and 200 mg/kg/dose oral ivermectin once daily on day 1, 2, 15 and 16 with full recovery and no further episodes of meningitis. The presented case underlines several critical points concerning the management of poorly known neglected diseases such as strongyloidiasis and HTLV infection in low-endemic areas. Despite several admissions for meningitis and strongyloidiasis, the parasitic infection was not adequately treated and the patient was not previously tested for HTLV. The supply of ivermectin and the choice of treatment scheme was challenging since ivermectin is not approved in Italy and there are no standardized guidelines for the treatment of severe strongyloidiasis in HTLV seropositive subjects

Monitoring Anti-PEG Antibodies Level upon Repeated Lipid Nanoparticle-Based COVID-19 Vaccine Administration

PEGylated lipids are one of the four constituents of lipid nanoparticle mRNA COVID-19 vaccines. Therefore, various concerns have been raised on the generation of anti-PEG antibodies and their potential role in inducing hypersensitivity reactions following vaccination or in reducing vaccine efficacy due to anti-carrier immunity. Here, we assess the prevalence of anti-PEG antibodies, in a cohort of vaccinated individuals, and give an overview of their time evolution after repeated vaccine administrations. Results indicate that, in our cohort, the presence of PEG in the formulation did not influence the level of anti-Spike antibodies generated upon vaccination and was not related to any reported, serious adverse effects. The time-course analysis of anti-PEG IgG showed no significant booster effect after each dose, whereas for IgM a significant increase in antibody levels was detected after the first and third dose. Data suggest that the presence of PEG in the formulation does not affect safety or efficacy of lipid-nanoparticle-based COVID-19 vaccines

Use of Miltefosine in a Patient With Mucosal Leishmaniasis and HIV-coinfection: A Challenge in Long-Term Management

The management of mucosal leishmaniasis in immunocompromised patients is not standardized and limited data are available on the use of miltefosine for treatment and secondary prophylaxis. We describe a case of mucosal leishmaniasis in an HIV-coinfected patient treated with miltefosine due to a severe allergic reaction to liposomal amphotericin B

Cytomegalovirus Infection Is Associated with Development of Chronic Lung Allograft Dysfunction

Background Cytomegalovirus (CMV) is the major and most common opportunistic infection complicating lung transplant (LTX). The aim of this study was to analyse the epidemiological aspects of CMV infection in lung transplant patients subject to a pre-emptive anti-CMV approach and to study the impact of this infection on lung transplant outcome, in terms of onset of chronic lung allograft dysfunction (CLAD).Methods This single-centre retrospective study enrolled 87 LTX patients (median age 55.81 years; 41 females, 23 single LTX, 64 bilateral LTX). All patients were managed with a pre-emptive anti-CMV approach. The incidences of the first episode of CMV infection, 1, 3, 6 and 12 months after LTX, were 12.64%, 44.26%, 50.77% and 56.14%. A median interval of 41 days elapsed between LTX and the first episode of CMV infection. The median blood load of CMV-DNA at diagnosis was 20,385 cp/ml; in 67.64% of cases, it was also the peak value. Patients who had at least one episode had shorter CLAD-free survival. Patients who had three or more episodes of CMV infection had the worst outcome.Results CMV infection was confirmed to be a common event in lung transplant patients, particularly in the first three months after transplant. It had a negative impact on transplant outcome, being a major risk factor for CLAD. The hypothesis that lower viral replication thresholds may increase the risk of CLAD is interesting and deserves further investigation. © 2022, The Author(s)

Evolution of macrolide resistance in streptococcus pyogenes over 14 years in an area of central Italy

We evaluated temporal fluctuations in macrolide resistance rates, analysing genetic determinants of resistance and clonal evolution in a population of 2744 S. pyogenes isolates collected in the period 2000–2013. The total resistance rate to erythromycin of the isolates was 17.9%. A maximum of erythromycin resistance emerged in 2000 (38.6%), followed by a significant decrease to 5.2% in 2012 (P<0.0001). Molecular analysis revealed the presence and co-presence of known genetic resistance determinants mefA, mefE, ermTR and ermB, in line with phenotypes. PFGE analysis identified genetically related groups in 2000 and 2007–2008, mainly the MLS and M phenotypes, respectively. The most prevalent emm types among a representative subset of resistant isolates were emm2, emm75 and emm77. All emm2 and 88.2% of the strains harbouring the emm75 gene were only recorded in M-phenotype strains, whilst all emm77-positive strains had the inducible MLS phenotype. The analysed susceptible isolates showed several emm types partially shared with resistant ones. Our results suggest that changes in bacterial population clonality, rather than horizontal transfer of resistance determinants, plays a major epidemiological role in S. pyogenes. Continuous monitoring of microbiological epidemiology seems to be crucial for correct and effective management of streptococcal infections

Trajectory of Spike-Specific B Cells Elicited by Two Doses of BNT162b2 mRNA Vaccine

: The mRNA vaccines for SARS-CoV-2 have demonstrated efficacy and immunogenicity in the real-world setting. However, most of the research on vaccine immunogenicity has been centered on characterizing the antibody response, with limited exploration into the persistence of spike-specific memory B cells. Here we monitored the durability of the memory B cell response up to 9 months post-vaccination, and characterized the trajectory of spike-specific B cell phenotypes in healthy individuals who received two doses of the BNT162b2 vaccine. To profile the spike-specific B cell response, we applied the tSNE and Cytotree automated approaches. Spike-specific IgA+ and IgG+ plasmablasts and IgA+ activated cells were observed 7 days after the second dose and disappeared 3 months later, while subsets of spike-specific IgG+ resting memory B cells became predominant 9 months after vaccination, and they were capable of differentiating into spike-specific IgG secreting cells when restimulated in vitro. Other subsets of spike-specific B cells, such as IgM+ or unswitched IgM+IgD+ or IgG+ double negative/atypical cells, were also elicited by the BNT162b2 vaccine and persisted up to month 9. The analysis of circulating spike-specific IgG, IgA, and IgM was in line with the plasmablasts observed. The longitudinal analysis of the antigen-specific B cell response elicited by mRNA-based vaccines provides valuable insights into our understanding of the immunogenicity of this novel vaccine platform destined for future widespread use, and it can help in guiding future decisions and vaccination schedules