Evaluation of three novel azasterols against Toxoplasma gondii

Previous studies from our group have demonstrated the high susceptibility of Toxoplasma gondii tachyzoites to the sterol analogues 22,26-azasterol and 24,25-(R,S)-epiminolanosterol. In this work we present data on testing in vitro three novel azasterols as potential agents for the treatment of toxoplasmosis. The three compounds inhibited parasite growth at micromolar concentrations, in a dose-dependent manner. Electron microscopy analysis of intracellular tachyzoites after treatment with the most effective compound showed drastic mitochondrion swelling associated with the appearance of an electron-lucent matrix and disrupted cristae. Parasite lysis also took place. The appearance of electron dense cytoplasmic structures similar to amylopectin granules distributed throughout the parasite suggests that azasterols might be inducing differentiation of those tachyzoites which were not lysed to the bradyzoite stage.     

Soft tissue sarcoma in dogs: A treatment review and a novel approach using electrochemotherapy in a case series

Canine soft tissue sarcomas (STSs) are locally invasive mesenchymal neoplasms. Electrochemotherapy (ECT) is an antitumour local ablative treatment that uses electric pulses to enhance the intracellular delivery of cytotoxic drugs. The aim of this retrospective study was to review the current treatment for STSs and to evaluate the efficacy and safety of ECT with bleomycin in canine STSs. Fifty‐two dogs with 54 STSs were included. Three groups were arranged: (a) ECT alone, (b) intra‐operative ECT and (c) adjuvant ECT. Signalment, tumour size, location, histological grade and margins and ECT parameters were collected. Recurrence rate (RR) and disease‐free interval (DFI) were calculated. Treatment toxicity was assessed using a 6‐point scale. STSs were mostly located on limbs (77.8%). Median tumour size was 4.3 cm (range 0.4‐17.0 cm). Most STSs were grade I (47.7%) and II (50.0%), and histological margins were incomplete in 94.5% of cases. Two complete remissions, one partial remission and one stable disease were recorded in group 1. Group 2 and 3 were similar for tumour location, size and grade, histological margins, treatment toxicity, pulse frequency and voltage. Moreover, RR and DFI were similar between group 2 and 3 (23% and 25%, 81.5 and 243 days, respectively). Local toxicity post ECT was mild (score ≤ 2) in 66.7% of cases. Higher toxicity score was associated with higher pulse voltage (1200 vs 1000 V/cm) (P = 0.0473). ECT coupled with bleomycin resulted safe and efficient in tumour local control and should be considered as an option for treatment of canine STSs.     

Outbreak of swimmer's itch in Central Italy: Description,causative agent and preventive measures

Swimmer's itch is caused by the penetration of free‐swimming larvae of trematodes of the family Schistosomatidae in human skin. It is usually reported in people engaged in recreational water activities in freshwater bodies and in most of cases, it is provoked by bird schistosomes of the genus Trichobilharzia. In the summer 2017, many cases of dermatitis were recorded in people bathing in the waters of the Albano Lake (Rome, Italy) and a parasitological investigation was carried out in order to ascertain the causative agent of these cases. Snails of the family Lymnaeidae, natural intermediate hosts of bird schistosomes, were collected from lake shallow waters to detect the presence of trematodes of the genus Trichobilharzia. Pools of maximum 10 snails were placed in Petri dishes, and cercarial emergence was stimulated exposing snails to strong artificial light intensity at 25°C. Three hundred and thirty‐seven snails were collected and screened for the shedding of cercariae. Furcocercariae of the family Schistosomatidae, with a morphology overlapping that of the genus Trichobilharzia, were detected in seven Petri dishes. Assuming that in each positive Petri dish just one snail was shedding furcocercariae, the minimum infectious rate was 2.1%. Molecular analysis of furcocercariae allowed ascribing them to the species Trichobilharzia franki. Snails of the species Radix auricularia were identified as intermediate hosts of the parasite. This is the second record of T. franki causing cercarial dermatitis in Central Italy, the third in Italy. The 2017 was in Italy exceptionally warm and dry. Trematodes are sensitive to changes in temperature, being cercarial production and emission rates temperature dependent. Small increases in water temperature would speed up parasite development and transmission, leading to a manifold increase in cercarial emergence. Moreover, high temperatures raise chances to acquire the infection, due to increased time spent in water by people.     

Leaf morphology and chemistry in Fagus sylvatica (beech) trees as affected by site factors and ozone: results from CONECOFOR permanent monitoring plots in Italy

During summer 2001, leaf samples were collected from seven beech stands that are part of the Italian network of permanent monitoring plots (CONECOFOR). In each plot, sun leaves from the upper crown were collected from five trees and subjected to chemical analysis (C, N, P, S, K, Ca and Mg) and morphological analysis (area, dry mass, thickness and thickness of individual tissues). Based on the measurements, nutrient ratios (N/C, N/K, N/P, N/Ca, N/Mg, K/Ca, Mg/Ca), sclerophylly indices (leaf mass per area (LMA) and leaf density (LD)) and nitrogen content per leaf area unit (NLA) were determined. Stomatal density was also measured. Among stands, leaf area was smallest and sclerophylly (indicated as LMA) was greatest in the southernmost stands and under drought conditions. Reduced leaf area and increased LMA also appeared to be strongly related to tropospheric ozone concentrations, whereas crown transparency was related mainly to site factors such as rainfall and temperature and, to a lesser extent, ozone concentration. The southernmost stands had a lower N/C ratio than the more northern stands, suggesting that the apoplastic fraction of cells within the leaf played a greater role. In the northern stands (especially at Lombardy and in Piedmont) where ozone concentrations were high, nutrient ratios were unbalanced and the high value of LMA appeared to be related primarily to the contribution of plasmatic components. Overall, leaf morphology was most sensitive to climate stress at the southern plots and to environmental pollution (nitrogen deposition and tropospheric ozone concentration) at some of the northern plots.     

Fate of enrofloxacin in swine sewage

The fate of enrofloxacin present in raw sewage at a swine-breeding facility was investigated by liquid-liquid extraction and reversed-phase liquid chromatography with photodiode array detection. Samples were collected in the storage pits of each pigsty and in a nonaerated lagoon used to stock the sewage. In the pigsties, the sewage was mixed with 10% olive oil groundwater, following a certified procedure (cod. Cer 020301) which reduces the bad odors and produces a better manure. This sewage treatment for 8 months in the lagoon dramatically reduced the amount of enrofloxacin to levels under the detection limit of 0.6 microg.L(-)(1). The results stress the importance of correct sludge management in limiting the impact of enrofloxacin in the environment.     

Elimination of in vitro bacterial contaminants in shoot cultures of ‘MRS 2/5’ plum hybrid by the use of <Emphasis Type="Italic">Melia azedarach</Emphasis> extracts

The antimicrobial activity of leaf and callus extracts of Melia azedarach was tested on in vitro shoot cultures of the peach rootstoch ‘MRS 2/5’ (Prunus cerasifera × Prunus spinosa) that were heavily contaminated with Sphingomonas paucimobilis (Sp) and Bacillus circulans (Bc). The extracts were filter-sterilised and added at 0%, 1%, 5%, 10% and 20% to a modified Murashige and Skoog proliferation medium previously autoclave-sterilised. Up to about 17% shoots died with 10–20% extract, except for Sp-contaminated shoots, whose survival was reduced to 50% after treatment with 20% extract. No shoots died with 1% to 5% supplement. The undiluted leaf extract showed bactericidal activity on plated Sp and Bc isolates. The homogenates of shoots randomly collected from treated cultures were processed for bacterial colony counting. Thus the 10% supplement was the best treatment for ridding Bc-contaminated cultures of bacteria (although 5% had a similar bactericidal effect), and allowing shoot growth and proliferation comparable to controls at the fifth subculture on a standard medium, while 20% extract was needed to eliminate Sp, and could induce higher growth and proliferation rates in surviving shoots than in untreated cultures. Callus extract was ineffective. The bactericidal activity of the leaf extract seemed attributable to a synergistic effect of azadirachtin with other unidentified compounds present in the extract.     

Identification of candidate genes for paratuberculosis resistance in the native Italian Garfagnina goat breed

Paratuberculosis disease is a chronic bacterial disease infection of ruminants of global relevance, caused by MAP (Mycobacterium avium subsp. paratuberculosis). The present study was conducted on the Garfagnina goat breed that is an Italian native goat population registered on the Tuscan regional repertory of genetic resources at risk of extinction. Forty-eight adult goats (27 serologically positive to MAP-positive and 21 serologically negative to MAP-negative) belonging to a single flock that had experienced annual mortalities due to MAP infection were identified and genotyped with the Illumina GoatSNP60 BeadChip. Diagnosis was achieved by serological tests, as well as post-mortem examination of affected animals. A genome-wide scan was then performed on the individual marker genotypes, in an attempt to identify genomic regions associated with MAP infection disease. Nine significant markers were highlighted and they were located within, or nearby, annotated genes. Two genes found in this study encode are linked to protein kinases that are among the most important enzymes involved in the immune response to Johne’s disease, and four genes are involved in the functions of the Golgi complex.