Determination of high molecular mass polycyclic aromatic hydrocarbons in a typical Italian smoked cheese by HPLC-FL

High-performance liquid chromatography with fluorescence detection was used for the analysis of polycyclic aromatic hydrocarbons (PAHs), in "Diavoletto" smoked cheese. Such cheese is typically produced in the Sorrento peninsula, and it is smoked commonly with different materials of vegetable origin. The importance of the smoking generation material is proven by the attention that the EU is paying in indicating the list of wood that may be used to produce smoking flavor agents. The PAHs considered are classified as "probable human carcinogens" by the U.S. Environmental Protection Agency for sufficient data from animal bioassays. The smoked samples contained high molecular mass PAHs with different levels ranging from 0.12 to 6.21 microg/kg. The determination was carried out also on liquid smoking flavor agents, smoke-flavored cheese, and nonsmoked cheese to measure the level of contamination before the treatment.     

Impact of Land Use and Urban Runoff on the Contamination of the Sarno River Basin in Southwestern Italy

A study was conducted to determine the contaminantlevel of the Sarno River basin, a small creek located insouthwestern Italy. Six sampling site locations were selected tocoincide with two previous studies conducted in 1975 and 1985 toprovide a basis for comparison. For each location, twelve sampleswere collected over the period October 1997–September 1998.Results indicated a degradation in river water quality, especiallynear the mouth. This was reflected by a dissolved oxygenconcentration of about 0.8 mg L^-1 near the urban areas ofScafati, Pompei and Castellamare of Stabia. At the same locations,BOD₅ and COD values were two to five times higher than theallowable regulatory maximum levels, respectively. About 4 kmfrom the headwaters, the average oxygen concentration was 4.7 mgL^-1, BOD₅ values were close to the standard limit (37 vs 40 mg L^-1), and COD values were above the standard limits(252 vs 160 mg L^-1). Highest concentrations were detectedduring peak tomato production. During this time, settleable solidswere also elevated, two to six times the regulatory standard.Coliform bacteria densities were found in excess of the standardlimits at all sites. Near the head of the river, mean fecalcoliform (22,571 MPN mL^-1) and streptococci densities (14,214 MPN mL^-1) surpassed the regulatory level of 120 and 20 MPN mL^-1, respectively, reflecting the input from the urbansettings of Sarno and agricultural fields. The heavy metaldissolved concentrations were low at all sampling sites. Most ofthe analyzed contaminants increased when comparend with 1985data. BOD₅ increased 5 to 10 times at sites near the urbansettings of Scafati, Pompei and Castellamare di Stabia, whereasCOD increases were twofold at all sites. A decline of chromiumlevels was observed due to the recent effective treatment ofwaste water from the tanning plants.     

Clostridium perfringens toxin-types in lambs and kids affected with gastroenteric pathologies in Italy

A study was carried out in the South of Italy to assess the role of clostridia in neonatal diseases of lambs and kids. Eighty-seven lambs and 15 kids belonging to 25 flocks were examined and Clostridium perfringens was the microorganism most commonly identified. C. perfringens isolates were analysed by polymerase chain reaction (PCR), in order to determine the prevalence of the genes cpa, cpb, cpb2, etx, iap and cpe. The most prevalent toxin-type of C. perfringens was found to be type A found in 84% of the cases with clostridial enterotoxaemia. No C. perfringens type B, C or E were found. C. perfringens type D was isolated in 16% of the cases. About 24% of the isolates were cpb2 positive. The prevalence of cpb2 across the different C. perfringens types varied. The beta(2)-toxin gene cpb2 was detected in 4/21 (19%) type A isolates, in 1/2 type D isolates, and in 1/2 type DE (cpe-carrying type D) isolates. The high rate of positivity to cpb2 among the isolates suggests that a vaccine based on the beta(2)-toxin, should be included in the vaccination schedule of the animals to confer adequate protection and to prevent the disease.     

Hydrogen peroxide localization and antioxidant status in the recovery of apricot plants from European Stone Fruit Yellows

Hydrogen peroxide (H₂O₂) localization and roles of peroxidases, malondialdehyde and reduced glutathione were compared in leaves of apricot (Prunus armeniaca) plants asymptomatic, European Stone Fruits Yellows (ESFY)-symptomatic and recovered. Nested PCR analysis revealed that Candidatus Phytoplasma prunorum, is present in asymptomatic, symptomatic and recovered apricot trees, confirming previous observations on this species, in which recovery does not seem to be related to the disappearance of phytoplasma from the plant.H₂O₂was detected cytochemically by its reaction with cerium chloride, which produces electron-dense deposits of cerium perhydroxides. H₂O₂was present in the plasmalemma of the phloem cells of recovered apricot plant leaves, but not in the asymptomatic or symptomatic material. Furthermore, by labelling apricot leaf tissues with diaminobenzidine DAB, no differences were found in the localization of peroxidases.Protein content in asymptomatic, symptomatic and recovered leaves was not significantly different from one another. In contrast, guaiacol peroxidase activity had the following trend: symptomatic > recovered > asymptomatic, whereas reduced glutathione content followed the opposite trend: asymptomatic > recovered > symptomatic. Moreover, no differences were observed in malondialdehyde concentrations between asymptomatic, symptomatic and recovered leaves. The overall results suggest that H₂O₂ and related metabolites and enzymes appear to be involved in lessening both pathogen virulence and disease symptom expression in ESFY-infected apricot plants.     

Species composition of Gasterophilus spp. (Diptera, Oestridae) causing equine gastric myiasis in southern Italy: parasite biodiversity and risks for extinction

Horse gastrointestinal myiasis caused by larvae of Gasterophilus spp. (Diptera, Oestridae) flies has a worldwide distribution and, where present, it is primarily caused by larvae of Gasterophilus intestinalis and Gasterophilus nasalis. Other species, i.e., Gasterophilus inermis, Gasterophilus pecorum and Gasterophilus haemorrhoidalis, present in different or in the same regions of the gastrointestinal tract, were only occasionally reported in very limited areas of eastern European Countries and in central Italy. With the aim to contribute data on the species composition of Gasterophilus and on the seasonal variation of the infection pattern in southern Italy, 152 native horses were necropsied from January to December 2003 and Gasterophilus larvae were collected from the stomach, the small intestine and the rectum of each of them. On the whole, 125 (82.2%) horses were infected by larvae of Gasterophilus spp. and 214 second stage larvae (L2) and 13,342 third stage larvae (L3) collected. Five species of Gasterophilus were identified with the following prevalence: G. intestinalis=95.2%, G. nasalis=44.8%, G. inermis=15.2%, G. pecorum=2.6% and G. haemorrhoidalis=0.8%. L3 were retrieved throughout the observation period with the highest mean burdens from January to August 2003 while the lowest mean was registered from September to November 2003. L2 were collected in February-March 2003 and from September to December 2003. The majority of the animals (n=66, 43.4%) were infected with a single Gasterophilus species, however, 45 animals (29.6%) harboured 2 species, 12 animals (7.9%) 3 species and 2 animals (1.3%) 4 species. The trend of abundance in the L3 of G. intestinalis and G. nasalis, the most represented species, was highly concordant (r=0.5, p<0.001). A retrospective comparison of our results and of other data from four seasons of observation (1983-1986) in central Italy showed that the number of G. inermis, G. pecorum and G. haemorrhoidalis have been decreasing relative to G. intestinalis and G. nasalis which may suggest tendency toward the extinction of the three former species of Gasterophilus.     

Intracranial epidural mucocele in a cat

An 18-month-old, spayed female, domestic shorthaired cat was presented with clinical signs of depression and reluctance to walk, which progressed to nonambulatory tetraparesis. Increased opacification of both frontal sinuses and a cyst-like abnormality causing compression and displacement of the right frontal lobe were seen on computed tomography. Bilateral frontal sinus trephination and right transfrontal craniotomy revealed clear, viscous fluid in the right frontal sinus and rostral fossa, compatible with an intracranial mucocele. At a 6-month follow-up examination, no signs of recurrence were appreciated.     

Histophagous scuticociliatids (Ciliophora) parasitizing turbot Scophthalmus maximus: morphology, in vitro culture and virulence

Systemic ciliatosis caused by histophagous ciliates constitutes a serious disease of cultured turbot. Six ciliate isolates were obtained from parasitized turbot during six epizootics at four different farms located in Spain, France and Portugal. Axenic cultures of the six isolates were obtained by periodical subculturing in ATCC 1651MA or supplemented L-15 media. In basal media or seawater, the parasites could survive starving for long periods with no apparent proliferation. In adequate media, growth kinetics was found to be very similar for isolates A and B, with a clear influence of temperature. Morphological studies demonstrated that all isolates share common features that allows their assignment to either Philasterides Kahl, 1931 or Miamiensis Thompson et Moewus, 1964. However, statistically significant differences were evident in pairwise comparisons of the isolates from the four farm sites in 16 taxonomically relevant morphometric features. This could allow the discrimination of different species or strains. Virulence of isolates A and B for healthy turbot was tested in several experiments. Differences in the virulence were especially evident after long-term in vitro culturing, isolate A being clearly attenuated after 35-42 passages, whereas isolate B became more virulent after 20-42 passages. The need of further studies to confirm such virulence variability and its implications in pathogenesis and prevention of turbot scuticociliatoses is stressed.     

Note: A comparison of molecular diagnostic procedures for the detection of aster yellows phytoplasmas (16Sr-I) in leafhopper vectors

Different molecular procedures were compared for the detection of aster yellows phytoplasmas (AYP) in the leafhopper vectorsMacrosteles quadripunctulatus (Kirschbaum),Euscelidius variegatus (Kirschbaum) andEuscelis incisus (Kirschbaum). Polymerase chain reaction (PCR) with universal and group-specific primers designed on the 16S-rDNA sequence was most sensitive in nested assays. A dot-blot procedure with an oligoprobe designed on the 16S-rDNA was less sensitive and consistent to detect phytoplasmas in total insect DNA, but consistently detected amplicons from direct PCR. The dot-blot assay with a probe based on a phytoplasma plasmid sequence detected AYP in most vector specimens and did not react with DNAs from leafhoppers infected by flavescence dorée and psyllids infected by apple proliferation phytoplasmas. This last assay is almost devoid of contamination risks, faster and cheaper compared to PCR, therefore it has to be preferred for field-scale analysis of leafhopper populations. http://www.phytoparasitica.org posting Feb. 24, 2004.     

Cross-transmission studies with Hypoderma lineatum de Vill. (Diptera: Oestridae): attempted infestation of goats (Capra hircus)

The potential for cross-transmission of Hypoderma lineatum from cattle to domestic goats (Capra hircus) was examined using artificial infestation techniques. Two routes of infestation, subcutaneous injection and dermal penetration, were used to expose goats to newly hatched first instars. Presence of antibodies and appearance of circulating antigen (hypodermin C) were evaluated at selected intervals for up to 40 weeks post-infestation. In addition, immunoblots against H. lineatum first-instar proteins were conducted using sera taken at 10 weeks post-infestation. Goats were palpated for the presence of developing larvae at sub-dermal sites beginning at week 30 pi. No developing larvae were palpated at any time, regardless of the route of infestation nor was circulating antigen detected in any infested goats. Antibodies were present at weeks 6 and 10 and week 27 pi in both infested groups. Immunoblots indicated all infested goats produced antibodies to first instar H. lineatum antigens. H. lineatum appears to be incapable of completing development in domestic goats although the transient appearance of ELISA detectable antibodies and the presence of bands on immunoblots suggests that at least some larvae survive long-enough to engender a humoural response. The host specificity of H. lineatum is discussed in light of the general concepts of host-parasite relationships of oestrids.     

Tuta absoluta,a South American pest of tomato now in the EPPO region: biology,distribution and damage

Tuta absoluta (Meyrick) is an invasive pest of tomato native to South America, where it is responsible for extensive damage. It rapidly spread into several European countries, becoming a key pest. Knowledge of its main biological traits can be used to develop effective plant protection management strategies. It is a multivoltine species with a homodynamous behaviour, and the length of its life cycle depends on environmental conditions, particularly temperature. The larvae feed and develop inside tomato leaves, stems and fruits throughout the entire growing cycle. The adults have crepuscular habits, and in Mediterranean conditions they can be easily detected throughout the year. Damage is directly related to the reduction of plants' photosynthetic capacity and of production levels in both protected and open‐field tomato crops; indirect damage can be also caused by secondary infections, with pathogens developing on the infested plant and fruit tissues.