Intraspecific variability of growth and ochratoxin A production by Aspergillus carbonarius from different foods and geographical areas.

Ochratoxin A (OTA) is a nephrotoxic mycotoxin naturally found in a wide range of food commodities throughout the world. Aspergillus carbonarius is the most important source of OTA in food commodities such as wine, grapes and dried vine fruits and is also responsible for the formation of OTA in coffee. The aim of this study was to determine the simultaneous effect of three culture media (Czapek Yeast Extract Broth (CYB); Synthetic Grape Juice Medium (SGM) and White grape juice (WGJ)) at three water activity (aw) levels (0.90; 0.95 and 0.98-0.99), and three incubation temperatures (15 °C, 25 °C and 35 °C) on the growth and OTA production by 16 strains of A. carbonarius. The strains were mainly isolated from grapes from areas with a Mediterranean climate. All the strains were confirmed for identity by sequencing of the calmodulin gene. The assay was performed in microtiter plates, determining the absorbance at 530 nm and the concentration of OTA after 1, 2, 4 and 10 days of incubation. No significant differences were observed in absorbance values between the strains. The highest absorbance values were recorded in CYB at 0.99 aw and at 0.95 aw after 10 days of incubation at 25 °C and 35 °C. None of the strains were able to grow at 0.90 aw and 15 °C in any culture media after 10 days of incubation. OTA concentration was statistically higher at 15 °C than at 25 °C or 35 °C. The highest significant OTA values were obtained at 0.98-0.99 aw and the best culture media for OTA production was CYB, followed by WGJ and SGM. While strains isolated from Mediterranean climate foods had a similar behavior despite being isolated from different geographical areas, OTA concentration produced by one Robusta coffee strain from Thailand was statistically higher at 25 °C than at 15 °C. This would suggest that the type of food matrices and consequently the adaptation of A. carbonarius strains to different climatic conditions would have a greater influence on the ecophysiology of the strains than only their geographical origin.

Impact of some environmental factors on growth and ochratoxin A production by Aspergillus niger and Aspergillus welwitschiae.

Ochratoxin A (OTA) is a nephrotoxic mycotoxin which may contaminate various foods and feed products worldwide. Aspergillus niger is one of the species responsible for OTA contamination in grapes and derived products. This species has recently been split into A. niger and Aspergillus welwitschiae. Both species can not be distinguished by phenotypic or extrolite profiles and to date there is no ecophysiological information of A. welwitschiae. The aim of this study was to determine the effects of water activity (aw) (0.90; 0.95 and 0.98-0.99), culture media (Yeast Extract Sucrose Broth (YESB); Synthetic Grape Juice Medium (SGM); White grape juice (WGJ)) and temperature (15 °C, 25 °C and 35 °C) on the growth and OTA production of four strains of A. niger and six strains of A. welwitschiae. The assay was performed in microtiter plates, determining the absorbance at 530 nm and the concentration of OTA at 1, 2, 4 and 10 days. No significant differences were observed in absorbance and OTA values between the two species under study. The highest absorbance values were recorded in YESB, followed by SGM and WGJ. Absorbance values increased with increasing aw and temperature. The highest OTA values were obtained at 0.98-0.99 aw and the best culture media for OTA production was YESB, followed by WGJ and SGM. The studied strains of A. niger produced the highest mean OTA level at 25 °C whereas A. welwitschiae strains produced the highest mean OTA concentration at 15 °C, although not differing significantly from concentration produced at 25 °C. To our knowledge, this is the first report on the impact of some environmental factors on growth and OTA production by A. welwitschiae.

Trichophyton erinacei in pet hedgehogs in Spain: Occurrence and revision of its taxonomic status.

Hedgehogs have increased in popularity as pets in Spain but there are no data of infection rates of this exotic animal with dermatophytes in our country. During the period of 2008-2011 a total of 20 pet hedgehogs (19 African pygmy hedgehogs and 1 Egyptian long-eared hedgehog) suspected of having dermatophytoses were studied. This is the first survey of the occurrence of T. erinacei in household hedgehogs in Spain. The T. erinacei infection rate was 50% (9 out of 19 African pygmy hedgehogs, and the one Egyptian long-eared hedgehog surveyed). Morphological identification of the isolates was confirmed by molecular analysis. All the strains had the same ITS sequence and showed 100% sequence similarity to T. erinacei type strain CBS 511.73 (AB 105793). The Spanish isolates were confirmed as T. erinacei urease positive. On the basis of ITS sequences, T. erinacei is a species close to but separate from the taxa included in the A. benhamiae complex. Review of the current literature on DNA-based methods for identification of species included in this complex has highlighted the urgent need to reach a consensus in species circumscription and classification system accepted by all mycologists.

Exploring the Capacity of Water Framework Directive Indices to Assess Ecosystem Services in Fluvial and Riparian Systems: Towards a Second Implementation Phase.

We explored the capacity of the biological and hydromorphological indices used in the Water Framework Directive (WFD) to assess ecosystem services by evaluating the ecological status of Spanish River Basins. This analysis relies on an exhaustive bibliography review which showed scientific evidence of the interlinkages between some ecosystem services and different hydromorphological and biological elements which have been used as indices in the WFD. Our findings indicate that, of a total of 38 ecosystem services analyzed, biological and hydromorphological indices can fully evaluate four ecosystem services. In addition, 18 ecosystem services can be partly evaluated by some of the analyzed indices, while 11 are not related with the indices. While Riparian Forest Quality was the index that was able to assess the largest number of ecosystem services (N = 12), the two indices of macrophytes offered very poor guarantees. Finally, biological indices related to diatoms and aquatic invertebrates and the Fluvial Habitat Index can be related with 7, 6, and 6 ecosystem services, respectively. Because the WFD indices currently used in Spain are not able to assess most of the ecosystem services analyzed, we suggest that there is potential to develop the second phase of the WFD implementation taking this approach into consideration. The incorporation of the ecosystem services approach into the WFD could provide the framework for assess the impacts of human activities on the quality of fluvial ecosystems and could give insights for water and watershed management in order to guarantee the delivery of multiple ecosystem services.

A biological tool to assess flow connectivity in reference temporary streams from the Mediterranean Basin.

Many streams in the Mediterranean Basin have temporary flow regimes. While timing for seasonal drought is predictable, they undergo strong inter-annual variability in flow intensity. This high hydrological variability and associated ecological responses challenge the ecological status assessment of temporary streams, particularly when setting reference conditions. This study examined the effects of flow connectivity in aquatic macroinvertebrates from seven reference temporary streams across the Mediterranean Basin where hydrological variability and flow conditions are well studied. We tested for the effect of flow cessation on two streamflow indices and on community composition, and, by performing random forest and classification tree analyses we identified important biological predictors for classifying the aquatic state either as flowing or disconnected pools. Flow cessation was critical for one of the streamflow indices studied and for community composition. Macroinvertebrate families found to be important for classifying the aquatic state were Hydrophilidae, Simuliidae, Hydropsychidae, Planorbiidae, Heptageniidae and Gerridae. For biological traits, trait categories associated to feeding habits, food, locomotion and substrate relation were the most important and provided more accurate predictions compared to taxonomy. A combination of selected metrics and associated thresholds based on the most important biological predictors (i.e. Bio-AS Tool) were proposed in order to assess the aquatic state in reference temporary streams, especially in the absence of hydrological data. Although further development is needed, the tool can be of particular interest for monitoring, restoration, and conservation purposes, representing an important step towards an adequate management of temporary rivers not only in the Mediterranean Basin but also in other regions vulnerable to the effects of climate change.

Six-Year Follow-up of Slaughterhouse Surveillance (2008-2013): The Catalan Slaughterhouse Support Network (SESC).

Meat inspection has the ultimate objective of declaring the meat and offal obtained from carcasses of slaughtered animals fit or unfit for human consumption. This safeguards the health of consumers by ensuring that the food coming from these establishments poses no risk to public health. Concomitantly, it contributes to animal disease surveillance. The Catalan Public Health Protection Agency (Generalitat de Catalunya) identified the need to provide its meat inspectors with a support structure to improve diagnostic capacity: the Slaughterhouse Support Network (SESC). The main goal of the SESC was to offer continuing education to meat inspectors to improve the diagnostic capacity for lesions observed in slaughterhouses. With this aim, a web-based application was designed that allowed meat inspectors to submit their inquiries, images of the lesions, and samples for laboratory analysis. This commentary reviews the cases from the first 6 years of SESC operation (2008-2013). The program not only provides continuing education to inspectors but also contributes to the collection of useful information on animal health and welfare. Therefore, SESC complements animal disease surveillance programs, such as those for tuberculosis, bovine cysticercosis, and porcine trichinellosis, and is a powerful tool for early detection of emerging animal diseases and zoonoses.

Aeromonas dhakensis pneumonia and sepsis in a neonate Risso's dolphin Grampus griseus from the Mediterranean Sea.

A neonate Risso's dolphin Grampus griseus was found stranded alive on a beach in Catalonia, Spain. Rehabilitation attempts were unsuccessful and it died 2 d later, showing pneumonia and sepsis. A pure bacterial culture was obtained from all tissues and blood and identified as Aeromonas hydrophila using the API 20NE. However, sequencing the rpoD gene showed that the strain in fact belongs to A. dhakensis, making this the first report of fatal haemorrhagic-necrotizing pneumonia and sepsis due to this species in a marine mammal. The A. dhakensis strain GMV-704 produced ß-haemolysis, possessed several virulence genes and showed sensitivity to several antimicrobials. This study provides a new potential host for A. dhakensis, and its potential virulence in dolphins and its presence in the marine environment may warrant considering this species a potential threat to marine mammals.

A new in vitro method to detect growth and ochratoxin A-producing ability of multiple fungal species commonly found in food commodities.

The aim of the study was to develop a new screening method to detect growth and ochratoxin A (OTA) production by multiple fungi growing in a small quantity of culture media, using microtiter plates. Eight ochratoxigenic species were included in the study. The strains were inoculated in sterile 96-well flat-bottom microtiter plates containing Yeast Extract Sucrose broth and Czapek Yeast Extract broth and incubated at 25 °C. Growth was daily monitored by absorbance measurements for 4 days and extended to 7 and 10 days for Penicillium spp. The entire experiment was repeated twice on different days. On each sampling time, five of the seven replicate wells inoculated for each strain and culture media were randomly selected and the content of each well was removed, extracted and injected into the HPLC. No statistically significant differences were observed for absorbance and OTA values, neither between replicates nor between experiments. Quantifiable OTA levels were detected after 48 h of incubation in Aspergillus alliaceus, Aspergillus carbonarius and Aspergillus niger, after 72 h in Aspergillus flocculosus, Aspergillus steynii and Aspergillus westerdijkiae and after 7 days in Penicillium nordicum and Penicillium verrucosum. The method offers the necessary tools for a rapid detection of growth and OTA production avoiding the use of plate cultures and can be very useful when many fungal isolates need to be screened.

Mycobiota and mycotoxin contamination of maize flours and popcorn kernels for human consumption commercialized in Spain.

Mycobiota and co-occurrence of aflatoxins, citrinin, ochratoxin A and zearalenone in 30 samples of maize flours and 30 of popcorn kernels purchased in Spain for human consumption were determined. The mycotoxin-producing ability of Aspergillus, Fusarium and Penicillium spp. was also studied. Total fungal counts of maize flours ranged from <10 to 8.4 × 10(4) CFU/g and predominant mycobiota belonged to Aspergillus spp. and Penicillium spp. In popcorn kernels samples the most frequent species were Aspergillus spp., Mucorales, Fusarium spp. and Penicillium spp. Aflatoxins were produced by Aspergillus flavus and Aspergillus parasiticus, citrinin by Penicillium citrinum and Penicillium verrucosum, ochratoxin A by Aspergillus niger and patulin by Aspergillus clavatus and Penicillium griseofulvum. Identification of all the mycotoxin-producing strains as well as some Aspergillus spp. difficult to identify using phenotypic characters only was also performed by molecular methods. Aflatoxins were detected in 14 maize flours and 2 popcorn kernels samples, while ochratoxin A was detected in 4 maize flours and 10 popcorn samples. Co-occurrence of aflatoxins and ochratoxin A was found in the 4 ochratoxin-positive maize flour samples. Citrinin and zearalenone were not detected. This is the first report of aflatoxins and ochratoxin A contamination in maize flours and popcorn kernels commercialized in Spain.

Pyogranulomatous pleuropneumonia and mediastinitis in ferrets (Mustela putorius furo) associated with Pseudomonas luteola Infection.

Between 2008 and 2009, three pet ferrets from different sources presented with acute episode of dyspnoea. Cytological examination of pleural exudates revealed severe purulent inflammation with abundant clusters of rod-shaped microorganisms with a clear surrounding halo. Treatment was ineffective and the ferrets died 2-5 days later. Two ferrets were subjected to necropsy examination, which revealed pyothorax, mediastinal lymphadenopathy and multiple white nodules (1-2mm) in the lungs. Microscopical examination showed multifocal necrotizing-pyogranulomatous pleuropneumonia and lymphadenitis with aggregates of encapsulated microorganisms, some of which were positively stained by periodic acid-Schiff and alcian blue. In-situ hybridization for Pneumocystis spp., Ziehl-Neelsen staining and immunohistochemistry for distemper, coronavirus and influenza antigen were negative in all cases. Electron microscopically, the bacteria were 2-3 µm long with a thick electron-lucent capsule. Microbiology from one ferret yielded a pure culture of gram-negative bacteria identified phenotypically as Pseudomonas luteola. This speciation was later confirmed by 16S RNA gene amplification.

[Comparative analysis between breath test, serological immunoassay and rapid-urease test for detection of Helicobacter pylori infection in Mexican patients with non-investigated dyspepsia]. / Un análisis comparativo entre prueba de aliento, serología y prueba de ureasa rápida para la detección de infección por Helicobacter pylori en pacientes mexicanos con dispepsia no investigada.

INTRODUCTION: There are multiple Helicobacter pylori (Hp) detection tests, some are invasive and other noninvasive. The diagnostic accuracy of these methods varies according to the prevalence of the disease. OBJECTIVE: To determine the diagnostic accuracy of the breath test, serology and rapid urease test, considering gastric biopsy with Giemsa stain as the gold standard in Hp-infected subjects with uninvestigated dyspepsia. METHODS: Eighty four subjects (64 women, mean age 45 years) who were referred for dyspeptic symptoms were evaluated. Also, 20 healthy volunteers (12 men, average age 38 years) were evaluated. All the subjects underwent hystological analysis with Giemsa stain, breath test (Heliprobe®), rapid urease test (CLOtest®) and serological immunoassay (Hexagon®). RESULTS: Overall, Hp infection was diagnosed by histological analysis in 59 subjects (49 patients and 10 healthy subjects). Positivity to breath test, rapid urease test and serology were 56%, 46% and 44% respectively. Agreement with the histological analysis was 0.902 for the breath test, 0.620 for rapid urease test and 0.45 for serology. The area under the curve for the breath test was 0.95, for the rapid urease test was 0.82 and for serological test was 0.74. CONCLUSIONS: In our population, the breath test shown to have a diagnostic accuracy equivalent to histological analysis by Giemsa in subjects with uninvestigated dyspepsia.

Effect of water activity, temperature and incubation time on growth and ochratoxin A production by Aspergillus niger and Aspergillus carbonarius on maize kernels.

The aim of this study was to determine the effects of water activity (a(w)) (0.92-0.98), temperature (5-45 °C) and incubation time (5-60 days) on growth and ochratoxin A (OTA) production by Aspergillus niger and Aspergillus carbonarius on maize kernels using a simple method. Colony diameters of both strains at 0.92 a(w) were significantly lower than those at 0.96 and 0.98 a(w) levels. The optimum growth temperature range for A. niger was 25-40 °C and for A. carbonarius 20-35 °C. A. niger produced OTA from 15 to 40 °C, and the highest OTA level was recorded at 15 °C. The concentration of OTA produced at 0.92 a(w) was significantly lower than those at 0.96 and 0.98 a(w). A. carbonarius produced OTA from 15 to 35 °C and the maximum concentration was achieved at 15 °C, although not differing statistically from the concentration detected at 20 °C. At 0.98 a(w) the OTA concentration was significantly higher than at 0.96 and 0.92 a(w). Our results show that maize supports both growth and OTA production by A. niger and A. carbonarius. The studied strains were able to produce OTA in maize kernels from the fifth day of incubation over a wide range of temperatures and water availabilities. Although the limit of quantification of our method was higher than that required for the analysis of OTA in food commodities, it has proved to be a useful and rapid way to detect OTA production by fungi inoculated onto natural substrates, in a similar way as for pure culture. Both species could be a source of OTA in this cereal in temperate and tropical zones of the world.

Temperature and incubation time effects on growth and ochratoxin A production by Aspergillus sclerotioniger and Aspergillus lacticoffeatus on culture media.

AIMS: As there is no knowledge of the influence of abiotic factors on the two new ochratoxin A (OTA)-producing species Aspergillus sclerotioniger and Aspergillus lacticoffeatus, the aim of this study was to evaluate the effect of temperature and incubation time on growth and OTA production by these species on culture media. METHODS AND RESULTS: The study was carried out on yeast extract sucrose agar (YES) and Czapek yeast extract agar (CYA) incubated at ten different temperatures from 5 to 50°C (at 5°C intervals). Growth assessment and OTA production were determined after 5, 10, 15, 20 and 30 days of incubation at each temperature. Aspergillus sclerotioniger grew from 10 to 35°C; OTA was detected from 10 to 35°C and the highest concentration was achieved at 15°C in CYA. Aspergillus lacticoffeatus grew from 10 to 45°C; OTA was detected from 15 to 45°C, and the maximum concentration was produced after 5 days at 25°C in YES. CONCLUSIONS: The studied species can produce OTA over a wide range of temperatures and significant amounts can be produced in only 5 days. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the influence of ecophysiological factors on these two ochratoxigenic species. The pattern of effects of temperature on growth and OTA production by A. sclerotioniger and A. lacticoffeatus was similar to those reported for the closely related species Aspergillus carbonarius and Aspergillus niger, respectively. The two new OTA-producing species have both been isolated from coffee beans, and the closely related ochratoxigenic species of section Nigri, A. carbonarius and A. niger are important sources of OTA in this substrate.

Osteoradionecrosis: an update.

Osteoradionecrosis (ORN) of the mandible is the most serious and severe side effect of combined treatment of head and neck tumors. A new theory for the pathogenesis of ORN has been proposed relating it to a fibro-atrophic mechanism including free radical formation, endothelial dysfunction, inflammation, microvascular thrombosis leading to bone and tissue necrosis. Risk factors mainly include radiation related risk factors, surgery and, tobacco and alcohol abuse. Removing of diseased teeth after and even probably after radiotherapy is generally considered the main risk factor in ORN. Conversely, steroid use before or after radiation may have a protective effect related to the inhibition of the initial inflammatory phase of ORN. Prevention of ORN is still based on the preventive extractions of decayed or periodontally compromised teeth before radiotherapy. Based on the current understanding of ORN pathophysiology, new preventive and therapeutic protocols have been suggested for mild to moderate stages. Free tissue surgical transfers is the treatment of choice of severe, extensive and long established ORN.

Bisphosphonate-related osteonecrosis of the jaws: how to manage cancer patients.

Bisphosphonate related osteonecrosis of the jaw (BRONJ) is defined as exposed necrotic bone appearing in the jaws of patients treated by systemic IV or oral BPs never irradiated in the head and neck area and that has persisted for more than 8 weeks. More than 90% of cases of osteonecrosis of the jaw have been in patients with cancer who received IV-BPs. The estimate of cumulative incidence of BRONJ in cancer patients with IV-BPs ranges from 0.8% to 18.6%. The pathogenesis of BRONJ appeared related to the potent osteoblast-inhibiting properties of BPs which act by blocking osteoclast recruitment, decreasing osteoclast activity and promoting osteoclast apoptosis. Dental extractions are the most potent local risk factor. Cancer patients wearing a denture could also be at increased risk of BRONJ. Non-healing mucosal breaches caused by dentures could be a portal for the oral flora to access bone, while the oral mucosa of patients on IV-BPs could also be defective. Whether periodontal disease is a risk factor for BRONJ remains controversial. Preventive measures are fundamental. Nevertheless, some teams have questioned its cost-effectiveness. The perceived limitations of surgical therapy of BRONJ led to the restriction of aggressive surgery to symptomatic patients with stage 3 BRONJ. The evidence-based literature on BRONJ is growing but there are still many controversial aspects.

Chrysosporium guarroi sp. nov. a new emerging pathogen of pet green iguanas (Iguana iguana).

Chrysosporium guarroi sp. nov. represented by five strains isolated from cases of dermatomycosis in pet green iguanas (Iguana iguana) in Spain, is described and illustrated. This taxon is characterized by its ability to grow at temperatures from 15 to 37 degrees C and by the presence of arthroconidia and aleurioconidia. The latter are unicellular, smooth, pyriform or clavate, sessile or borne at the ends of narrow stalks. The analysis of the sequences of the D1/D2 and ITS regions confirm the separation of this new species from others of the genus Chrysosporium.

[Oral health: social determinants of a health inequality]. / Santé buccale: déterminants sociaux d'un terrain majeur des inégalités.

The oral health of disadvantaged social groups is worse at all the ages than that of the favored groups. If tooth decay prevalence decreases, this disease is still unequally distributed: 20% of the children, those with the weakest socio-economic statute (SES), concentrate 60% of the decays. Edentulism strikes significantly more people with weak SES. The inequalities of oral health reflect those of general health. Evidence of the inequalities in oral health is exposed even in the developed countries. Different models of intervention are presented: risk groups identification and targeting by specific programs; oral health community approach which includes socio-economic and public health measures aiming all the population; insurance approach to be combined with the preceding ones.

Occupational changes in manual tactile sensibility of the dentist.

The aim of this study was to find out if the use of vibrating tools (drill, ultrasonic scaler ...) in dental practice has negative side effects on the manual tactile sensibility of the dentist. The sensory tests were performed on 50 subjects, who were allocated to three different groups according to their occupation and the length of their working experience. The first test group consisted of 20 dentists, who had more than 25 years of work experience. As a control group, 20 non-dentists were recruited to obtain a similar age distribution as the former test group. A final group consisted of 10 dentists with more than 1 year of work experience. All subjects underwent three tests: light-touch sensation test, two-point discrimination test and thermal sensation test on the thumb and the index finger of the working and non-working hands. Results showed significant differences, especially for the light-touch sensation test. The study showed more specifically that the tactile sensibility of the working hand of the dentists with more than 25 years of work experience was significantly diminished with respect to the non-working hand and to the working hand of non-dentists. In the test group of young dentists, there was no noticeable reduction of manual tactile sensibility.

Dermatomycosis in a pet inland bearded dragon (Pogona vitticeps) caused by a Chrysosporium species related to Nannizziopsis vriesii.

A Chrysosporium sp. related to Nannizziopsis vriesii was isolated in pure culture from squames and biopsies of facial lesions in a pet inland bearded dragon (Pogona vitticeps) in Spain. The presence in histological sections of morphologically consistent fungal elements strongly incriminates this fungus as the aetiological agent of infection. Lesions regressed following treatment with oral ketoconazole and topical chlorhexidine and terbinafine until the lizard was lost to follow up 1 month later. The ITS-5.8S rRNA gene of the isolate was sequenced and a search on the GenBank database revealed a high match with the sequences of two Chrysosporium sp. strains recently isolated from green iguanas (Iguana iguana) with dermatomycosis, also in Spain. Phylogenetic analysis of the sequences revealed that all these strains are related to N. vriesii. This is the first report of dermatomycoses caused by a Chrysosporium species related to N. vriesii in a bearded dragon outside North America.