Effect of Coffee Silver Skin and Brewers' Spent Grain in the Control of Root-knot Nematodes.

Plant parasitic nematodes (PPN) are important pests of numerous agricultural crops especially vegetables, able to cause remarkable yield losses correlated to soil nematode population densities at sowing or transplant. The concern on environmental risks, stemming from the use of chemical pesticides acting as nematicides, compels to their replacement with more sustainable pest control strategies. To verify the effect of aqueous extracts of the agro-industry waste coffee silverskin (CS) and brewers' spent grain (BSG) on the widespread root-knot nematode Meloidogyne incognita, and on the physiology of tomato plants, a pot experiment was carried out in a glasshouse at 25 ± 2 °C. The possible phytotoxicity of CS and BSG extracts was assessed on garden cress seeds. Tomato plants (landrace of Apulia Region) were transplanted in an artificial nematode infested soil with an initial population density of 3.17 eggs and juveniles/mL soil. CS and BSG were applied at rates of 50 and 100 % (1L/pot). Untreated and Fenamiphos EC 240 (nematicide) (0.01 µL a.i./mL soil) treated plants were used as controls. Reactive oxygen species (ROS) and chlorophyll content of tomato plants were estimated during the experiment. CS extract, at both doses, significantly reduced nematode population in comparison to the untreated control, although it was less effective than Fenamiphos. BSG extract did not reduce final nematode population compared to the control. Ten days after the first treatment, CS 100 %, BSG 50 % and BSG 100% elicited the highest ROS values, which considerably affected the growth of tomato plants in comparison to the untreated plants. The control of these pests is meeting with difficulties because of the current national and international regulations in force, which are limiting the use of synthetic nematicides. Therefore, CS extracts could assume economic relevance, as alternative products to be used in sustainable strategies for nematode management.

Prevalence and genotypic characterization of Clostridium difficile from ruminants in Switzerland.

Clostridium difficile is an emerging enteric pathogen of humans and animals with a known main reservoir in the intestinal tract of various warm-blooded animals. This study was carried out to evaluate the prevalence of C. difficile in 150 rectal swab samples collected from cattle and goats in Switzerland. The overall prevalence of C. difficile was 6.6%. The isolates belonged to the PCR ribotypes 033, 066, 070, 003, 001 and 137. In addition, the occurrence of C. difficile in faecal samples collected from farm ground was also evaluated and C. difficile was detected in 21% (7/30) of these samples. These isolates belonged to ribotypes 033, 066, 014 and 137. Several isolates pertained to ribotypes known to cause Clostridium difficile infections in humans.

Occurrence of toxigenic Clostridium difficile in edible bivalve molluscs.

Clostridium difficile is an anaerobic bacterium commonly considered to be responsible for antibiotic-associated gastrointestinal diseases, ranging from diarrhea of varying severity to pseudomembranous colitis. The aim of this study was to assess the occurrence of C. difficile in marine edible bivalve molluscs, which, as filter feeding organisms, are able to accumulate particles suspended in water, including microorganisms. Samples of Mytilus galloprovincialis, Tapes philippinarum, and Venus verrucosa were collected from mussel farms and fishmongers in the province of Naples (Southern Italy). C. difficile was found in 49% of the 53 samples investigated. Sixteen isolates were grouped in 12 known different PCR ribotypes (001, 002, 003, 010, 012, 014/020, 018, 045, 070, 078, 106, and 126), whereas 10 additional isolates were grouped in 8 new PCR riboprofiles. Two toxinotypes (0 and V) were found. Fifty eight percent of the isolates were toxigenic. These findings indicate that toxigenic C. difficile strains can be isolated in bivalve molluscs. Marine filter feeding organisms, therefore, may be considered as reservoir of toxigenic strains of C. difficile. The ingestion of raw or poorly cooked contaminated seafood and the high temperature resistance of the spore-forming C. difficile could represent an important source of exposure and pose human health concern.

Phenotypic characterization and putative virulence factors of human, animal and environmental isolates of Plesiomonas shigelloides.

Plesiomonas shigelloides (a bacterium widely distributed in aquatic ecosystems causing both intestinal and extra-intestinal diseases) shows a host of putative virulence markers, such as hemolysins, cytotoxins, production of exoenzymes associated with pathogenicity, adhesive ability and vacuolation of cell lines in vitro. Technical difficulties in detecting some of these virulence factors together with scantiness of epidemiological information, due to the lack of routine analysis for P. shigelloides as etiological agent of gastroenteritis, lead to sporadic and occasional finding of these bacteria. All this casts doubt on the real virulence potential of P. shigelloides and fuels a debate about assignment of these bacteria to the list of human pathogens. Here we demonstrated the phenotypic diversity and the putative virulence markers by examining serotype biochemical and virulence properties of 60 strains of P. shigelloides isolated from human, animal and environmental samples in different countries, which showed the unpredictable occurrence of the above properties depending on various locations and diverse sources.

Incidence and characterisation of Aeromonas spp. in environmental and human samples in southern Italy.

The incidence and characterisation of Aeromonas species in human and environmental samples in southern Italy were investigated. The results emphasize that 12.3% of the 210 examined patients carried Aeromonas spp. in their faeces. These results underline the need to include Aeromonas spp. in the list of routinely analysed enteropathogens in all diarrhoeal stool samples, especially in children below 10 years of age, elderly persons and immunocompromised patients.

Extractable soil lipids and microbial activity as affected by Bt and non Bt maize grown on a silty clay loam soil.

Pyrolysis-gas (Py-GC) chromatography was used to characterize extractable lipids from Bt and non-Bt maize shoots and soils collected at time of harvesting. Py-GC-MS (mass spectrometry) showed that the concentrations of total alkenes identified in non-Bt shoots and soils were 47.9 and 21.3% higher than in Bt maize shoots and soils, respectively. N-alkanes identified were of similar orders of magnitude in Bt and non-Bt maize shoots, but were 28.6% higher in Bt than in non-Bt soils. Bt maize shoots contained 29.7% more n-fatty acids than non-Bt maize shoots, whereas the concentrations of n-fatty acids in Bt soils were twice as high as those in non-Bt soils. Concentrations of unsaturated fatty acids in Bt maize shoots were 22.1% higher than those in non-Bt maize shoots, while concentrations of unsaturated fatty acids were 22.5% higher in non-Bt than in Bt soils. The cumulative CO2-C evolved from soils under Bt and non-Bt crops was 30.5% lower under Bt as compared to non-Bt crops, whereas when maize shoots were added to Bt and non-Bt soils, the decrease in CO2-C evolved were 16.5 and 23.6%, respectively. Our data showed that the cultivation of Bt maize significantly increased the saturated to unsaturated lipid ratios in soils which appeared to negatively affect microbial activity.

The importance of pathogenic organisms in sewage and sewage sludge.

Deficient sanitation poses a serious threat to human and animal health, involving complex relationships between environments, animals, refuse, food, pathogens, parasites, and man. However, by sanitizing and stabilizing the organic matter of sewage sludge, agriculture can utilize it to maintain soil, water, and air quality. As ingredients in soil amendments, such bioresidues are a source of nutrients for plants. Stabilization and sanitation of sewage sludge safely couple its recycling and disposal. This coupling becomes increasingly important as economic and environmental constraints make strategies for waste disposal more difficult to apply. The occurrence of viruses, bacteria, yeasts, fungi, and zooparasites in sewage sludge is reviewed in this article, and consequential epidemiologic concerns that arise from sewage sludge recycling is also addressed.

Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy.

A survey was undertaken to examine sea water and sediment for the presence of Vibrio and Aeromonas spp. along approximately 900 km of coast in Southern Italy during early and late summer. A quantitative analysis was also done to evaluate the water fecal contamination at the stations examined. The results indicate that all the investigated areas were submitted to a wide spatial fluctuation of fecal contamination and that Vibrio and Aeromonas spp. were present in both high and low fecal-contaminated stations. Sixty two percent of the investigated samples were positive for Aeromonas spp., while 42% of samples were positive for Vibrio spp. It was interesting to note that 38% of the positive stations for both Aeromonas and Vibrio spp. showed a fecal coliform contamination of water at < 10(2) cells 100 ml(-1). Thus, these findings support the hypothesis that the bacterial indicators (such as fecal coliforms) do not always satisfactorily reflect the hygienic quality of water. The presence of Vibrionaceae on copepods was also investigated. Copepods were sampled at a station located inside the harbour of the city of Naples and were found contaminated by V. cholerae non-O1, V. alginolyticus, V. fluvialis and A. caviae. Furthermore, the antibiotic resistance patterns of isolated bacteria showed the presence of a number of resistant strains among the isolates. In order to discriminate the isolates on the basis of their biochemical profiles and/or antibiotic resistance patterns, cluster analysis was carried out which showed that no unique assay could fully discern these isolates. However, the best discrimination resulted from complete pattern profile based on both biochemical profiles and antibiotic resistance patterns.

Ecological relationship between Aeromonas and Vibrio spp. and planktonic copepods in the coastal marine environment in southern Italy.

The colonisation of planktonic copepod integument by bacteria belonging to the family of Vibrionaceae is a well described phenomenon. In this study, besides reporting on the occurrence of Vibrionaceae and other enteropathogens, we further report on the bacterial attachment to the Estuarine copepod Acartia margalefi in a faecal polluted coastal lagoon near Naples, Southern Italy. In addition, we also performed a laboratory experiment to study the ability of 7 bacterial strains (Vibrio cholerae non-Ol, V. mimicus, V. parahaemolyticus, V. alginolyticus, Aeromonas hydrophila, Escherichia coli and Pseudomonas sp.) to colonise the copepod integument. For this laboratory study, 4 different species of copepods, namely Temora stylifera, A. clausi, Centropages typicus and Paracalanus parvus sampled from the Gulf of Naples (Southern Italy) were used. Scanning electron microscope (SEM) studies on the copepods sampled from the lagoon indicated that the bacterial attachment on the integument of Acartia margalefi was mainly on the ventro-lateral body region of the host and in the joints of the thoracic segments, as well as on the swimming and feeding appendages. This infestation, made by rod-like bacteria, was absent in winter but reached peak values of 70% frequency in June. The laboratory studies showed that while V. cholerae non-O1 and A. hydrophila attached on live and dead copepods, respectively, the V. parahaemolyticus, V. alginolyticus, V. mimicus, E. coli and Pseudomonas sp. failed to colonise either live or dead copepods. Thus, this study provides further valuable information about the ecological relationship between different microorganisms (pathogens) and copepods in the coastal marine environment in Southern Italy.

Isolation, and virulence profiles, of Aeromonas hydrophila implicated in an outbreak of food poisoning in Sweden.

A case of food poisoning outbreak involving Aeromonas hydrophila is reported in this study. A group of 27 people consumed a typical Swedish food "landgång" which is a type of smörgåsbord containing shrimps with mayonnaise, liver paté, ham, sausage, and legume salad which was purchased from a food store. Twenty-two of the 27 persons became ill within 20-34 hr of consumption of the food and reported the symptoms ranging from severe acute diarrhea, abdominal pain, headache, fever and vomiting. One person also fainted. The symptoms lasted for a couple of days. Of the remaining 5 healthy persons who consumed the left-over food the next day, 2 became ill with similar symptoms. The bacteriological examination of left-over food samples resulted in the isolation of A. hydrophila from shrimps with mayonnaise, smoked sausage, liver paté and boiled ham. The total number of A. hydrophila in these foods were log 10(6) to log > 10(7) organisms per gram of food sample. A. hydrophila was however, not isolated from legume/mayonnaise salad samples. All the food samples tested showed low numbers of other expected food contaminating organisms such as coliforms at 37 C and 44 C, fecal streptococci, Staphylococcus aureus, fungi and yeast etc., while Bacillus cereus, Clostridium perfringens and Salmonella spp. were not detected in the food samples. Investigations of the virulence profiles of the A. hydrophila isolates showed their capacity to produce beta-hemolysin, cytotoxins, cytotonic toxins, enterotoxins, and adhesion to and invasion of human intestinal (Henle 407) cells in culture.

An Outbreak of Aeromonas hydrophila Infection in Turtles (Pseudemis scripta).

An outbreak of Aeromonas hydrophila infection with a high rate of mortality (95%) in turtles (Pseudemis scripta) in Italy is reported. Pure cultures of the pathogen were isolated from liver, lung, kidney, and heart specimens of the turtles. The A. hydrophila isolate was resistant to amoxicillin, ampicillin, cephalothin, and trimethoprim-sulfamethoxazole but was sensitive to a number of other antibiotics tested. The study indicates that pet turtles can act as reservoirs of this pathogen and may play an important role in the etiology of Aeromonas-associated human infections.

Comparison of putative virulence factors in Aeromonas hydrophila strains isolated from the marine environment and human diarrheal cases in southern Italy.

Aeromonas hydrophila strains isolated from the same geographical region (southern Italy) but from different sources (sea sediments and human diarrhea cases) were characterized for the production of potential virulence determinants, such as production of cytotoxins, cytotonic toxins, hemolysin, and dermonecrotic factors and their capacity to adhere to human intestinal 407 cells in vitro. The results showed that isolates from both the sources produced all or some of the virulence factors which may be involved in the pathogenesis of Aeromonas-associated infections. Our study indicates that further epidemiological studies are necessary to elucidate the public health significance of infections caused by Aeromonas spp.

Detection of potential virulence markers of Vibrio vulnificus strains isolated from fish in Sweden.

A variety of potential virulence markers such as the production of cytotoxin, haemolysin, exoenzymes, bactericidal action of sera, presence of capsule and adhesion to human intestinal cells were investigated on Vibrio vulnificus strains isolated from eels in Sweden. The strains had the capacity of producing all or some of the above-mentioned virulence markers, to varying degrees though none of the strains produced any capsule. The strains also bound specifically to human intestinal cells in vitro with maximum adhesion levels of 30 bacteria/cell. The results on binding of V. vulnificus cytotoxin to HeLa cells, showed that a very short exposure time (30 min) was required for inducing the cytotoxic effects. V. vulnificus is a relatively new addition to the list of bacteria pathogenic for humans, and since there are increasing reports on its isolation from aquatic environments and seafood (e.g. raw oysters, crabs and shellfish), the results on virulence profiles of V. vulnificus strains presented above emphasize the importance of these organisms in public health and epidemiological studies.

Fatal septicaemia caused by Aeromonas hydrophila in a patient with cirrhosis.

In this case report from Italy we describe a fatal infection caused by A. hydrophila in a 39 yr old cirrhotic patient. This pathogen was isolated as a pure single culture from the patient's blood sample. The patient died on the second day of hospitalization from overwhelming sepsis. The A. hydrophila isolate was tested for different potential virulence properties, such as invasiveness, adherence, exotoxins production, presence of fimbriae and for the patterns of resistance to a variety of antimicrobial agents. Although, the Aeromonas species are infrequently reported as a cause of human infections, the present case study confirms the capability of these pathogens to induce serious human infections.