Sperm ultrastructure and spermatogenesis in the Japanese beetle Popillia japonica (Coleoptera, Scarabaeidae).

Popillia japonica is an invasive scarab beetle native to Japan that in 1916 invaded New Jersey in USA. From that moment onwards, the insect has spread invading several US states, Canada, the Azores, Italy and, recently, Switzerland. It is a severe agricultural pest included in the EU priority pest list being able to feed on more than 300 plant species and having an important biotic potential. The general morphology of the reproductive apparatus shows paired testes, each of them having six testicular lobes grouped in threes. From the ventral part of each testicular lobe, each containing about 20 follicles, an efferent vessel originates that fuses with the other efferent vessels to form the deferent duct. A pair of long tubular accessory glands is present. The deferent ducts and accessory glands fuse together into an ejaculatory duct before entering the aedeagus. The sperm is a typical pterygote sperm, 110 µm long, composed of a head and a tail. In the head a three-layered acrosome of about 6 µm in length and a nucleus of about 18 µm long are present. During sperm maturation two C-shaped structures appear in the cytoplasm from the opposite sides of the nucleus that then disappear in late spermatids. In the tail a typical 9 + 9 + 2 flagellar axoneme and two mitochondrial derivatives are present. Moreover, in the head-tail transition region the centriolar adjunct forms a sheath from which three elongated accessory bodies originate. Two of these accessory bodies are placed alongside the axoneme, whilst the third one is placed beneath the mitochondrial derivatives. Mature sperm are grouped in cysts containing about 256 sperm cells. A morphological comparison with related species is provided.

Adaptive management of invasive pests in natural protected areas: the case of Matsucoccus feytaudi in Central Italy.

Invasive species are a significant threat to affected ecosystems, having serious environmental, economic and social impacts. The maritime pine bast scale, Matsucoccus feytaudi Ducasse (Hemiptera: Matsucoccidae), causes serious damage to Pinus pinaster forests in SE France, Corsica and Italy where it has been introduced. This study illustrates the adaptive management plan implemented in the Migliarino, San Rossore, Massaciuccoli Regional Natural Park in Tuscany, Italy, where M. feytaudi arrived in 2004, leading to the decay of local P. pinaster stands. The management programme, aimed at slowing the establishment and growth of M. feytaudi, was carried out in the main sector of the park, Tenuta di San Rossore, to retard the destruction of the P. pinaster coastal strip protecting the more internal woodland from sea salt and to allow replacement of P. pinaster trees with a more stable broad-leaved wood. The combined use of mass trapping and silvicultural interventions, applied in a targeted manner according to distribution maps of pest captures and damage, helped to delay forest destruction compared with a nearby unmanaged area of the park Tenuta di Tombolo. Although M. feytaudi continued to spread during the management period, the populations remained at low levels for 6 years, showing a marked increase in 2012. During this period, the P. pinaster stands were reduced from 320 to 249 ha. The final result of this ongoing gradual conversion process will be transformation of the P. pinaster forest into Holm oak woods and Mediterranean shrub land, while P. pinaster will survive as clusters or blocks of trees.

First Report of Botryosphaeria dothidea on Sycamore, Red Oak, and English Oak in Northwestern Italy.

The Parco Nord Milano, Italy is a 600-ha green area (45°53'71″N, 9°20'97″E). Since the spring of 2002, extensive and unknown decline was observed on 20-year-old sycamore (Acer pseudoplatanus), red oak (Quercus rubra), and English oak (Q. robur) plantations. The trees showed branch and twig dieback associated with bark cankering along the whole stem and an irregular to wilted crown. A closer inspection of the inner part of the symptomatic trunk and roots revealed a dark bluish tissue discoloration. From the symptomatic trunk and root tissue, a dark gray abundant mycelium was consistently isolated on potato dextrose agar (PDA). The sequenced internal transcribed spacer (ITS) regions of rDNA (GenBank Accession Nos. DQ198265, DQ198266, and DQ198267) showed 99% identity to Botryosphaeria dothidea strain CBS110302 (GenBank Accession No. AY259092). A Fusicoccum sp. with black pycnidia was consistently collected from the cankers. The conidia were hyaline, aseptate, fusiform to narrowly ellipsoidal, with a truncate base (22 × 4.5 µm), referred to as Fusicoccum aesculi (CBS identification), the anamorph of B. dothidea (2). Pathogenicity tests were conducted for all three hosts by stem inoculation on 18-month-old seedlings growing in plastic pots containing a 2:1 turf/sand mixture. Two experiments were conducted using two inoculation techniques. In the first trial, 6-mm-diameter mycelial plugs of B. dothidea were applied to 6-mm-long bark wounds. The same inoculation method was used for application to bark without wounding. Control seedlings were inoculated with sterile agar plugs in a similar fashion as above. Inoculated and control seedlings were kept in a growth chamber and watered once per week. In the second trial, segments of branches 15-cm long were inoculated with 6-mm-diameter B. dothidea plugs (1), with and without any wounding. Control segments of branches were treated with sterile agar plugs in place of fungal mycelium. The branches were incubated at 23°C in moist chambers. For both experiments, the inoculated stem portions were wrapped with Parafilm to prevent desiccation. There were three replicate seedlings per inoculation technique, experiment, and plant species. After 2 months, all seedlings showed bark cankers and pycnidial formation, while the controls were symptomless. An inner dark bluish stem tissue discoloration was observed. The symptoms were more abundant on the segment of branches where the inoculum was applied to the wounded bark. B. dothidea was successfully reisolated, confirming Koch's postulates. To our knowledge, although it is accepted as synonymous of B. berengeriana (2), this is the first report of B. dothidea on sycamore, red oak, and English oak in Italy. The fungus was previously reported in Italy to cause canker on Platanus spp. References: (1) M. E. Sanchez et al. Plant Dis. 87:1515, 2003. (2) B. Slippers et al. Mycologia 96:83, 2004.

Birth weight for gestational age centiles for Italian neonates.

OBJECTIVE: To provide centiles for birth weight (BW) according to gestational age (GA) and sex for infants born in Italy. METHODS: We used records of the whole neonatal population of Tuscany, a region in Italy, from July 1991 to June 2002 as resulting from the database of the cystic fibrosis neonatal screening program (n=290129). We excluded as unlikely for GA those BW that were more than two interquartile ranges above the 75th centile or below the 25th centile for each GA and gender group. RESULTS: We present the 3rd, 10th, 25th, 50th, 75th, 90th and 97th centiles of BW for GA from the 24th to 43rd week of gestation for male and female Italian neonates, as both tables and smoothed curves. CONCLUSIONS: The large size of the examined population allows us to provide up-to-date, reliable BW for GA centiles for Italian newborns, especially for lower GAs.

[Anti-Pseudomonas aeruginosa antibodies, circulating immune complexes, and anticytoplasm antibodies of neutrophils in patients with cystic fibrosis with and without Pseudomonas aeruginosa colonization]. / Anticorpi anti-Pseudomonas aeruginosa, immunocomplessi circolanti ed anticorpi anticitoplasma dei neutrofili in pazienti affetti da fibrosi cistica colonizzati e non da Pseudomonas aeruginosa.

Chronic lung infections, mainly due to Pseudomonas aeruginosa (Pa), account for the most of the morbidity and mortality in CF patients. The pathogenic factors predisposing to airway colonization are still nuclear. Host's immune response is not only poorly protective but can also act as a damaging factor in the development of the disease. Moreover, clinical manifestations of an overactive immune response, including vasculitis and arthropathy, have been recently described in the CF population. To deepen factors involved in the pathogenesis of lung injury, we evaluated the presence of anti-Pa precipitating antibodies, circulating immune complexes (CIC) and antineutrophil cytoplasmic antibodies (ANCA) in sera from 50 CF patients colonized and not by Pa. Number of anti-Pa precipitins was significantly different in chronically and in not colonized patients (p < 0.001; t = 7.75). Anti-Pa antibodies were positively correlated to age (p = 0.002, r = 0.42) and inversely correlated to lung function parameters (p = 0.031 r = -0.35 with respect to FVC). Mean C3-CIC levels in the sera were statistically higher in chronically colonized patients (p = 0.013; t = 2.57); while there was not a significant difference with respect to C1q-CIC values. Four patients, all chronically colonized by Pa, were ANCA-positive at indirect immunofluorescence, showing a cytoplasmic pattern. All 50 patients were found to be negative when tested for anti-mpo and anti-pr3 antibodies with ELISA. In conclusion, our data demonstrate that persistence of Pa provides a stimulus for chronic inflammation and the immune response in CF patients, leading to anti-Pa antibodies and CIC production. Even in the face of further research, we speculate that c-ANCA production is secondary to neutrophil activation with a consequent release of cytoplasmic enzymes that perpetuate and increase the inflammatory process. Since anti-mpo and anti-pr3 antibodies rarely occur in CF patients we suggest that other antigenic specificities may be involved in ANCA production.

Allergic bronchopulmonary aspergillosis in Italian cystic fibrosis patients: prevalence and percentage of positive tests in the employed diagnostic criteria.

The prevalence of allergic bronchopulmonary aspergillosis (ABPA) in cystic fibrosis (CF) patients is difficult to determine because the data in the literature are not homogeneous or comparable. ABPA and CF have similar clinical symptoms which make diagnosis difficult and underestimate the real dimensions of the problem. We conducted an epidemiological study on 3089 Italian CF patients to determine the prevalence of ABPA in Italy and verify the percentage of positive tests in the employed diagnostic criteria. Our results indicate that the prevalence of ABPA in Italian CF patients is 6.18%, mainly in adolescents and young adults. ABPA is diagnosed using clinical symptoms (presence of episodic bronchial obstructions or typical radiographic features) and on the basis of other criteria which can only be partially fulfilled in paediatric patients. Among the diagnostic tests the most sensitive are the total IgE (84.5%), specific IgE anti-Aspergillus fumigatus (81.6%) and the prick test (68.3%). In the absence of clinical symptoms and gold standard diagnostic tests, serological positivity and/or the skin test are not sufficient evidence to confirm the presence of ABPA.

Multiresistant non-fermentative gram-negative bacteria in cystic fibrosis patients: the results of an Italian multicenter study. Italian Group for Cystic Fibrosis microbiology.

We carried out an epidemiological study on 2717 patients seen on a regular basis at Italian Cystic Fibrosis Centers with the aim of defining the prevalence of multiresistant Gram-negative strains in Italy. We found 272 multiresistant strains out of 1560 Gram-negative strains isolated in 1012 Italian cystic fibrosis patients. From our results in Italian cystic fibrosis patients we may conclude: (1) the beta-lactam antibiotics are moderately active; ceftazidime is the most efficacious even if 59.9% of multiresistant strains are not sensitive to this drug; (2) the aminoglycosides are poorly efficacious; 93% of multiresistant strains are resistant in vitro to tobramycin; and (3) the quinolones, notwithstanding their relatively recent introduction into clinical practice, have very poor activity against multiresistant strains, 89.7% of which are not sensitive.

[Bacterial infections and resistance to antibiotics in cystic fibrosis]. / Infezioni batteriche e resistenza agli antibiotici nella fibrosi cistica.

Knowledge of the microbiology of pulmonary infections is critical for treatment of cystic fibrosis because sickness and mortality in this disease are mainly due to relapse occurring in the respiratory tract. The microbiology of pulmonary infections presents several singular aspects. Respiratory tract infections are caused by bacteria such as Staphylococcus aureus in the early years of life and Pseudomonas aeruginosa and Burkholderia cepacia thereafter. The patients, who are not immune compromised, are predisposed to chronic colonization and highly transmissible bacterial strains can cause cross-infections. Bacterial also develop resistance mechanisms which make them difficult to treat. Until recently the relationship between genetic defects and a predisposition to colonization was not noted, but recent studies have allowed us to form some interesting hypothesis. The present work analyzes the principal mechanisms of antibiotic resistance, with particular reference to classic cystic fibrosis pathogens, and looks at future prospects of respiratory tract infection treatment.

Evaluation of glucose tolerance and insulin secretion in cystic fibrosis patients.

UNLABELLED: Blood glucose profile and insulin levels during the oral glucose tolerance test were retrospectively analyzed in 52 cystic fibrosis (CF) patients aged 10-33 years (mean 18 years). Thirty-seven subjects (71.1%) had normal glucose tolerance; 6 patients (11.5%) had impaired glucose tolerance and 9 patients (17.3%), with fasting blood glucose levels below 7.2 mmol/l were classified as diabetic, according to WHO diagnostic criteria. No significant differences in the total amount of secreted insulin (expressed as area under the curve) were detected in CF patients with or without glucose intolerance, but a significant delay in the insulin peak time appeared to be related to a compromised nutritional status. Our data indicate a significantly increased risk of glucose metabolism impairment related to poor nutritional condition (RR 5.40; 95% CI: 1.5-19.7) and worse clinical status (RR 4.27; 95% CI: 1.05-17.2). In particular, glucose tolerance abnormalities were found in 50% of CF patients with an unsatisfactory nutritional condition versus 15.6% of CF patients with good nutritional status. CONCLUSIONS: Since CF-related diabetes is often underdiagnosed and associated with deterioration in patients' overall clinical status, it is very important to identify subjects at risk of developing diabetes by strict monitoring of glucose metabolism when deterioration in nutritional and clinical conditions is seen which cannot be otherwise explained.

Genetic history of cystic fibrosis mutations in Italy. I. Regional distribution.

Earlier analysis of the Italian population showed patterns of genetic differentiation that were interpreted as being the result of population settlements going back to pre-Roman times. DNA disease mutations may be a powerful tool in further testing this hypothesis since the analysis of diseased individuals can detect variants too rare to be resolved in normal individuals. We present data on the relative frequencies of 60 cystic fibrosis (CF) mutations in Italy and the geographical distribution of the 12 most frequent CF mutations screened in 3492 CF chromosomes originating in 13 Italian regions. The 12 most frequent mutations characterize about 73% of the Italian CF chromosomes. The most common mutation, delta F508, has an average frequency of 51%, followed by N1303K and G542X, both with average frequencies around 5%. Multivariate analyses show that the relative frequencies of CF mutations are heterogeneous among Italian regions, and that this heterogeneity is weakly correlated with the geographical pattern of non-DNA 'classical' genetic markers. The northern regions are well differentiated from the central-southern regions and within the former group the western and eastern regions are remarkably distinct. Moreover, Sardinia shows the presence of mutation T338I, which seems absent in any other European CF chromosome. The north-western regions of Italy, characterized by the mutation 1717-1G-->A, were under Celtic influence, while the north-east regions, characterized by the mutations R1162X, 2183AA-->G and 711 + 5G-->A, were under the influence of the Venetic culture.

[Pseudomonas aeruginosa antibodies in patients with cystic fibrosis: clinical implications]. / Anticorpi anti-Pseudomonas aeruginosa in pazienti con fibrosi cistica: implicazioni cliniche.

Patients chronically colonised by P. aeruginosa develop a pronounced antibody response against P. aeruginosa that can be used to discriminate between superficial colonization and chronic infection. Anti-P. aeruginosa antibodies fail to afford protection against this pathogen; moreover high levels of antibodies are correlated with a poor prognosis. We investigated the significance of anti-P. aeruginosa antibodies (precipitins) by crossed immunoelectrophoresis (CIE) in 94 patients attending the Cystic Fibrosis Center of Florence. The highest numbers of precipitins were found in serum from patients chronically colonized in comparison to those patients who were transiently or not colonized. A negative correlation was found between the number of precipitin peaks and clinical conditions, evaluated with Schwachman score, and the number of precipitins and pulmonary functions. In summary, anti-P. aeruginosa antibodies fail to protect against P. aeruginosa bronchopulmonary infections, and are correlated to a more severe disease. Based on our experience, P. aeruginosa antibodies can be considered a reliable index of lung damage.

[Azlocillin in the treatment of pulmonary infections in patients with cystic fibrosis: plasma concentrations and therapeutic indications]. / L'azlocillina nel trattamento delle infezioni polmonari dei pazienti affetti da fibrosi cistica: concentrazioni plasmatiche e indicazioni terapeutiche.

Azlocillin plasma concentrations have been studied in 10 cystic fibrosis patients suffering from chronic pulmonary infections with Pseudomonas aeruginosa. Patients were given single i.v. doses of 100 e 200 mg/kg body weight as intravenous infusion over 30 minutes. Azlocillin plasma levels have been assayed by a rapid, sensitive and precise high performance liquid chromatographic method. After the dose of 100 mg/kg body weight concentrations of azlocillin decreased below the therapeutic concentrations after three hours; dose of 200 mg/kg was followed by plasma concentrations in the therapeutically desirable range during the 6-8 hours study period. The pharmacokinetic analysis offers further evidence of the dose-dependent nature of azlocillin elimination. Higher dosage of 200 mg/kg body weight and monitoring of plasma drug levels are recommended in the therapy of patients with cystic fibrosis.

[Experience acquired over a 4-year period with the Vichi method in the study of bronchopulmonary changes in cystic fibrosis]. / Esperienza acquisita in un quadriennio con il metodo di Vichi nello studio delle alterazioni broncopolmonari della fibrosi cistica.

The classification of chest alterations in Cystic Fibrosis (CF) and related score proposed by Chrispin and Norman has been widely adopted in Europe and is still applied (although slightly modified) in most European Centres. Brasfield classification instead has been mostly used in the USA. Lately, however, to revise both classifications, the need has been felt, for a more precise correlation to anatomo-radiological data as well as for inclusion of headings which have not been taken into account so far. In 1980 one of the Authors (Vichi) worked out a new scoring system for the chest alterations of CF. Results are reported from a follow-up of 15 patients with CF carried out at the FC Centre of Meyer Ospedale in Florence from the late 1981 to 1985. The patients underwent periodic checkings including determination of clinical scoring system-according to Shwachman and Kulczychi modified by Doershuk-respiratory function tests, chest X-rays evaluated by three radiologists separately, following both Chrispin and Norman and Vichi scoring systems. The latter system has proved to be well correlated to clinical data and to the ordinary pulmonary function tests but it mainly presents a high observer reproducibility.

[Pharmacokinetics of oral theophylline. Comparison between asthmatic and cystic fibrosis patients]. / Farmacocinetica della teofillina orale. Confronto tra pazienti asmatici e pazienti con fibrosi cistica.

The pharmacokinetics of oral theophylline at steady-state were comparatively investigated in 13 asthmatic patients and in 10 patients with cystic fibrosis (CF). In all patients, the drug was administered twice daily as slow-release tablets. The total daily dose of theophylline ranged from 10.8 to 29.4 mg/kg/day. For each patient, the time-course of theophylline steady-state plasma levels was studied after the morning dose. Six serial plasma samples were drawn at 0, 2, 4, 6, 8, and 12 h after dosing. Model-independent methods were used for calculating the pharmacokinetics parameters (area under the curve and clearance). The clearance values (mean +/- SD) calculated in the two patient groups were significantly different (asthmatic patients: clearance = 61.2 +/- 15.6 ml/h/kg; CF patients: clearance = 86.3 +/- 22.8 ml/h/kg; P = 0.007). It has previously been shown that the clearance of theophylline after single dose is increased in CF patients. Our study confirms this finding under steady-state conditions and demonstrates that higher theophylline doses are on the average required to treat patients with FC.

[Our experience with antibiotic therapy in children hospitalized between 1979 and 1981]. / Esperienze sull'antibioticoterapia nel bambino ricoverato 1979-1981.

The use of antibiotics in the Children Hospital of Florence has been studied considering the whole hospital for 1979 and only one department for 1980-81. 7832 clinical records of children admitted to the Hospital have been analyzed in order to get more informations about the quality of antibiotic treatment and to understand the causes of use and misuse of this kind of drugs in pediatric practice.