Selected pathological, immunohistochemical and ultrastructural changes associated with an infection by Diphyllobothrium dendriticum (Nitzsch, 1824) (Cestoda) plerocercoids in Coregonus lavaretus (L.) (Coregonidae)

The pathological changes induced by an infection of Diphyllobothrium dendriticum (Nitzsch, 1824) plerocercoids in powan, Coregonus lavaretus (L.), from Loch Lomond, Scotland, were assessed using immunohistochemical and ultrastructural techniques. In a sample of 26 powan, the occurrence of encysted plerocercoids of D. dendriticum on the outer surface of the stomach was 38.5% (n = 10) with the number of cysts ranging from 4 to 15 and measuring 4.2 +/- 1.0 mm x 3.4 +/- 0.9 mm (mean +/- SD). Histological examination of intestinal samples also revealed plerocercoids (2-21) encapsulated within a proliferation of mesenteric fibrous tissues of the gastric wall and, occasionally, by the gut lamina propria-submucosa and lamina muscularis. In section, cysts were tri-layered and were formed from a series of concentric whorls of fibroblast and collagen fibre-based connective elements. The extent of necrosis within each muscle layer and the serosa of the stomach differed, notably within the latter that was marked by a chronic inflammatory reaction and fibrosis. Within the cyst and around it, a large number of degranulating mast cell/eosinophilic granule cells were seen, in addition to melano-macrophage centres. Immunohistochemical staining of sections of infected stomach revealed a high density of elements, in close proximity to plerocercoids, staining positive for serotonin, bombesin, substance P and galanin. Uninfected material did not present the same levels of activity. Sections through both infected and uninfected tissue were also tested for elements containing vasoactive intestinal peptide, met-enkephalin, calcitonin gene-related peptide, neuropeptide Y and nitric oxide synthase, but these were absent.     

Side effects of botanical insecticides on predatory mite Amblyseius andersoni (Chant)

The effects of the botanical insecticides Biopiren plus, Piresan plus and Neemazal T/S on the predatory mite Amblyseius andersoni (Chant) were tested under laboratory, semi-field, and field conditions. The tests carried out in the laboratory allowed detection of the direct toxicity on eggs and females as well as the effects on fecundity, whereas semi-field trials assessed their residual toxicity. In laboratory and semi-field tests, pyrethrins, particularly Piresan plus, proved to have the highest toxicity with E value, concerning both female mortality and fecundity, of 100?% and 45?%, respectively. The same product tested in the field only caused a significant reduction in the phytoseiid population shortly after the treatment. The relevant escape rate registered in semi-field tests could explain the lack of phytoseiid eradication in the field after treatment with pyrethrins.     

Apple cultivars acceptance by Dysaphis plantaginea Passerini (Homoptera: Aphididae)

The development time, longevity, and fecundity of Dysaphis plantaginea were measured on six apple cultivars (Golden delicious, Red delicious, Renetta canada, Florina querina, Golden lasa, and Golden orange) under controlled conditions, in order to establish the life tables of D. plantaginea on the different cultivars and to evaluate the tolerance of the cultivars against this pest. The intrinsic rates of increase (r _m) were 0.07 and 0.11 day⁻¹ on F. querina and G. orange, respectively. On the other four cultivars, rates ranged between 0.22 and 0.26 day⁻¹, with the maximum value registered on G. lasa. All the biological and demographic parameters evaluated, i.e. survival of larvae (M = 5.8–72.2%), mean generation time (T = 14.94–20.62 days), and net reproduction rate (R ₀ = 4.23–47.37), show that G. orange and F. querina are tolerant cultivars while R. delicious, R. canada, G. lasa, and G. delicious are susceptible hosts.     

A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria

A simple and rapid multiplexed sandwich chemiluminescent enzyme immunoassay has been developed for the simultaneous detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes. To achieve the multiplexed detection of the four pathogens, a new polystyrene 96 well microtiter plate format has been designed, in which each main well contains four subwells in the bottom. The monoclonal antibodies specific for each bacteria were separately immobilized in each subwell. When the samples were added to the main wells, the bacteria able to specifically bind to the corresponding monoclonal antibody were captured in one of the four subwells. Subsequently, a mixture of peroxidase-labeled polyclonal antibodies against the four bacteria was added and the peroxidase activity of the bound polyclonal labeled antibodies in each well was measured by an enhanced luminol-based chemiluminescent cocktail using a low-light charge-coupled imaging device. The assay was simple and fast, and the limit of quantification was in the order of 104-105 CFU/mL for all bacterial species. The accuracy of the method, evaluated by comparison of the results with a conventional culturing methodology, was satisfactory, with recovery values ranging from 90 to 120%. This method can be used as a screening test to evaluate the presence of these pathogen bacteria in different foodstuffs.     

One-step triplex-polymerase chain reaction assay for the authentication of yellowfin (Thunnus albacares), bigeye (Thunnus obesus), and skipjack (Katsuwonus pelamis) tuna DNA from fresh, frozen, and canned tuna samples

A one-step triplex-polymerase chain reaction (PCR)-based assay was developed to discriminate between three tuna species, Thunnus albacares, Thunnus obesus, and Katsuwonus pelamis, even in highly processed food samples such as canned or cooked tuna. Diagnostic nucleotides were identified by direct sequencing and alignment of part of the mitochondrial cytochrome b gene of 30 authenticated exemplars, which allowed us to evaluate intraspecific variation and the genetic distance between three tuna species. The assay relies on a one-step triplex-PCR reaction in which in a single tube species-specific amplification products are generated only in the presence of the correct template nucleic acid and the species of origin of the DNA is indicated by the distinctive size of the PCR product. The identification of tuna species can be performed with a good accuracy, low cost, and with potential automation for large-scale high-throughput screenings in small in-house laboratories.     

Antioxidant and antimutagenic properties of Hibiscus tiliaceus L. methanolic extract

The genus Hibiscus thrives in a variety of climates and produces a diversity of natural compounds with bioactive properties. We have studied the chemical composition and the in vivo antioxidant properties of Hibiscus tiliaceus L. methanolic flower extract, as well as its mutagenic/antimutagenic effects. Vitamin E and some stigmasterol derivatives that might confer an antioxidant effect to the extract were present. Treatment with this extract protected several Saccharomyces cerevisiae strains defective in antioxidant defenses against H2O2 and t-BOOH cytotoxicities, showing a clear antioxidant activity. The effect is the same for all strains used, independent of the antioxidant defense disrupted, suggesting that protection may be due to molecules that act as versatile and wide spectrum nonenzymatic antioxidants, such as vitamins or phytosterols. The extract was not mutagenic in either Salmonella typhimurium or S. cerevisiae and showed a significant antimutagenic action against oxidative mutagens in S. cerevisiae.     

Risk factors for highly pathogenic H7N1 avian influenza virus infection in poultry during the 1999-2000 epidemic in Italy

In 1999-2000, Italian poultry production was disrupted by an H7N1 virus subtype epidemic of highly pathogenic avian influenza (HPAI). The objectives of the present study were to identify risk factors for infection on poultry farms located in regions that had the highest number of outbreaks (Veneto and Lombardia) and the impact of pre-emptive culling as a complementary measure for eradicating infection. A Cox regression model that included spatial factors, such as the G index, was used. The results confirmed the relationship between risk of infection and poultry species, production type and size of farms. The effectiveness of pre-emptive culling was confirmed. An increased risk of infection was observed for poultry farms located near an infected farm and those at altitudes less than 150m above sea level. The measures for the control and eradication of AI virus infection need to consider species differences in susceptibility, the types of production and the density of poultry farms in the affected areas.