Antibiotic Resistance of Bacteria Isolated from the Internal Organs of Edible Snow Crabs

Antibiotic resistance and microbiota within edible snow crabs are important for the Chionoecetes (snow crab) fishing industry. We investigated these parameters using culture methods and antibiotic susceptibility tests with six internal organs from three species of Chionoecetes. Each sample revealed many unexpected microbial species within Chionoecetes internal organs. On the basis of 16S rRNA sequence analysis of 381 isolates, the most abundant genera identified in Chionoecetes opilio were Acinetobacter spp. (24%), Bacillus spp. (4%), Pseudomonas spp. (34%), Stenotrophomonas spp. (28%), and Agreia spp. (11%). In Chionoecetes sp. crabs, Acinetobacter spp. (23%), Bacillus spp. (12%), and Psychrobacter spp. (20%) were most prevalent, while Agreia spp. (11%), Bacillus spp. (31%), Microbacterium spp. (10%), Rhodococcus spp. (12%), and Agrococcus spp. (6%) were most abundant in C. japonicus. Our antibiotic resistance test found resistance to all nine antibiotics tested in 19, 14, and two of the isolates from C. opilio, Chionoecetes sp., and, C. japonicus respectively. Our results are the first to show that microbes with antibiotic resistance are widely distributed throughout the internal organs of natural snow crabs.     

Effect of indigenous mycobiota on ochratoxin A production by Aspergillus carbonarius isolated from soil

This study determined the biotic interaction between 30 non-toxigenic indigenous strains of Aspergillus niger aggregate, Aspergillus flavus, Trichoderma spp., Mucor spp., Cladosporium spp., Ulocladium spp., Curvularia spp., Absidia spp., Geotrichum spp. and Acremonium spp., isolated from soil destined for maize crops, with respect to their ability to prevent ochratoxin A (OTA) production by A. carbonarius on “in vitro” assay, on liquid and solid medium. OTA production was completely inhibited when A. carbonarius was inoculated in a interactive mixed culture with all A. niger aggregate strains assayed, a 80 % of Trichoderma spp. strains, a 40 % of Cladosporium spp. strains, Acremonium spp and Geotrichum spp; only one strain of A. flavus tested was able to completely inhibit the mycotoxin accumulation. OTA production increased when A. carbonarius ACS 8 was growing on liquid interactive mixed culture with Mucor spp strains. These results demonstrated that OTA production by Aspergillua carbonarius strain was significantly influenced by the presence of different non-toxigenic fungal strains when growing together on paired cultures.     

Simultaneous and rapid detection of enteric pathogens from raw milk by multiplex PCR

Enteroinvasive Escherichia coli (EIEC), heat-labile enterotoxin (LT) E. coli, Shigella spp., and Salmonella spp. are common enteric pathogens, which cause food-borne diseases if consumed in contaminated milk products. The rapid and reliable methods for detecting are imperative for reduction in hazard of infection. In this study, we selected primers, optimized the polymerase chain reaction (PCR) conditions, and analyzed the sensitivity and specificity of the multiplex PCR assay to screen raw milk from these enteric bacteria. Furthermore, EIEC, LT-E. coli, Shigella spp., Salmonella spp., and 11 non-targeted pathogenic strains were performed for the specificity of the multiplex PCR. Specific bands showed in EIEC, LT-E. coli, Shigella spp., and Salmonella spp. but no bands showed in other 11 pathogenic strains. The sensitivity of multiplex PCR was relatively high, was rounded to 200 CFU/ml (Shigella spp. and EIEC), 320 CFU/ml (Salmonella spp.), and 100 CFU/ml (LT-E. coli). This method for simultaneous and rapid detection of enteric pathogens (EIEC, LT-E. coli, Shigella spp., and Salmonella spp.) in raw milk showed high sensitivity and specificity, and led to faster track to report results.     

Stratigraphy and palynology of the Pennsylvanian continental Buçaco Basin (NW Iberia)

The Buçaco Basin is a Pennsylvanian continental basin located along an important NNW–SSE strike structure (Porto-Tomar-Ferreira do Alentejo shear zone) that separates the Ossa-Morena and Central Iberian Zones of the Iberian Variscan Fold Belt in central western Portugal. The shear zone controlled the sedimentation in the basin and probably its post-sedimentary evolution. Sedimentation is initially alluvial with characteristic red sandstones, breccias and conglomerates. A gradual change to a fluvial (and probably lacustrine) type of sedimentation is observed with finning-upward cycles of gravel conglomerates, sandstones and organic-rich mudstones with occasional coal seams. Three representative sections were sampled for palynology and seventeen samples yielded sporomorphs with moderate to poor preservation. The palynological content from the alluvial sediments shows low diversity and poorly preserved assemblages dominated by Triquitrites spp., Densosporites spp., Laevigatosporites spp., and other taxa associated with siliciclastic environments or rheophytic mires. The fluvial and lacustrine sediments show a dramatic increase in diversity with an abundant, typical peatland microflora including sporomorphs such as Endosporites spp., Lycospora spp. and Monoletes spp., but also marginal peat and siliciclastic substrate taxa such as Densosporites spp., Latensina/Cordaitina spp., and Florinites spp. Other common taxa are Cheiledonites spp., Crassispora spp., Dictyotriletes-like miospores (mostly fragments), Potonieisporites spp., and Wilsonites spp. The presence and considerable abundance of Potonieisporites novicus and Cheiledonites cf. major is indicative of the middle to upper Potonieisporites novicus-bhardwajii–Cheiledonites major (NBM) miospore biozone of Western Europe, corresponding to the late Stephanian (early Gzhelian).     

A survey of outdoor and indoor airborne fungal spora in the Redemption City, Ogun State, south-western Nigeria

The concentration and biodiversity of airborne fungi of the Redemption City, an immense campground for Christian faithful and the temporary site of the Redeemer’s University in south-western Nigeria, was studied between February and May 2011 using the culture plate method. The study was undertaken to assess the concentrations of fungal spores and their health implication in this ever-busy environment. Fifteen different sites classified as closed or open were selected. During the experiment, a total of 228 colonies were counted, and 29 fungal species belonging to 26 genera were isolated which include the following: Aspergillus flavus, Aspergillus niger, Bipolaris spp., Chrysosporium spp., Cladosporium spp., Coniothyrium corda, Curvularia spp., Diplodia spp., Fusarium spp., Gliocladium spp., Monilia spp., Mucor spp., Mucor plumbeus, Penicillium spp., Phycomyces spp., Phytophthora spp., Pilobolus spp., Pyrenochaeta spp., Rhizopus stolonifer, Torula spp., Trichoderma spp. and Trichophyton spp. The most frequently occurring fungi were A. niger, C. corda and M. plumbeus, while the least recorded were Torula and Trichophyton species. Majority of the fungi isolated are known allergens; they could also be opportunistic causing various diseases in man. There is therefore a dire need for good sanitation practices within the studied areas of the camp.     

The prevalence and diversity of intestinal parasitic infections in humans and domestic animals in a rural Cambodian village

In Cambodia, intestinal parasitic infections are prevalent in humans and particularly in children. Yet, information on potentially zoonotic parasites in animal reservoir hosts is lacking. In May 2012, faecal samples from 218 humans, 94 dogs and 76 pigs were collected from 67 households in Dong village, Preah Vihear province, Cambodia. Faecal samples were examined microscopically using sodium nitrate and zinc sulphate flotation methods, the Baermann method, Koga Agar plate culture, formalin-ether concentration technique and Kato Katz technique. PCR was used to confirm hookworm, Ascaris spp., Giardia spp. and Blastocystis spp. Major gastrointestinal parasitic infections found in humans included hookworms (63.3%), Entamoeba spp. (27.1%) and Strongyloides stercoralis (24.3%). In dogs, hookworm (80.8%), Spirometra spp. (21.3%) and Strongyloides spp. (14.9%) were most commonly detected and in pigs Isospora suis (75.0%), Oesophagostomum spp. (73.7%) and Entamoeba spp. (31.6%) were found. Eleven parasite species were detected in dogs (eight helminths and three protozoa), seven of which have zoonotic potential, including hookworm, Strongyloides spp., Trichuris spp., Toxocara canis, Echinostoma spp., Giardia duodenalis and Entamoeba spp. Five of the parasite species detected in pigs also have zoonotic potential, including Ascaris spp., Trichuris spp., Capillaria spp., Balantidium coli and Entamoeba spp. Further molecular epidemiological studies will aid characterisation of parasite species and genotypes and allow further insight into the potential for zoonotic cross transmission of parasites in this community.     

Representation of aquatic vegetation change by plant macrofossils in a small and shallow freshwater lake

We explored spatial and temporal relationships between contemporary aquatic vegetation and surface sediment macrofossil remains in a small, shallow, English lake (Green Plantation Pond). The aquatic vegetation of Green Plantation Pond underwent a marked compositional change after 2005 with a shift from Elodea spp.-Potamogeton pusillus-Chara spp. to Ceratophyllum spp.-Chara spp.-Potamogeton crispus dominance. By comparing macrophyte and plant macrofossil distributions at multiple, closely spaced points in Green Plantation Pond for 2000 and 2008–2009, we studied the ability of macrofossils to track this major aquatic vegetation change. Representation of macrophytes by macrofossils was high with 63 and 76 % of extant plant species recorded by macro-remains in the 2000 and 2009 sediment surveys respectively. Nevertheless, plants were both over-represented (Nitella flexilis, Chara spp. and Zannichellia palustris) and under-represented (Ranunculus sect. Batrachium, Potamogeton spp.) in the sediment record in terms of relative macrofossil abundances and the number of occupied sample points. The study also revealed a lack of preservation of Elodea spp. leaf remains in the second (2009) survey compared to the first (2000) probably due to a longer time interval (5 vs. 10 months) between macrophyte and sediment sampling. Nevertheless, the macrofossils reliably recorded both the main shift in the contemporary vegetation (e.g. especially increases in Ceratophyllum spp. and P. crispus abundance) and other more subtle floristic changes (e.g. increases in Myriophyllum spicatum and Lemna spp.) exceptionally well. This study highlights the huge potential of macrofossils for tracking sub-decadal changes in the aquatic vegetation of small, shallow lakes.     

Genetic diversity of tick-borne pathogenes in Tomsk and environs

Two urban and two suburban biotopes of Tomsk were studied for tick-transmitted disease prevalence in ticks collected in the wild. Tick-borne encephalitis virus (TBEV), West Nile virus (WNV), Borrelia spp., Rickettsia spp., and Ehrlichia spp. were found in 6.5%, 2.2%, 8%, 2.5%, and 1.7% of tick specimens, respectively. Genetic markers of Powassan virus, Bartonella spp., and Babesia spp. were not found. Analysis of the genetic diversity of revealed pathogens demonstrated that TBEV strains belonged to the Siberian and Far-Eastern subtypes, and the Far-Eastern subtype of TBEV is most frequently found in urban biotopes (up to 43% of urban strains of TBEV). WNV strains belonged to the 1a genotype. Borrelia spp. was classified as B. garinii, Rickettsia spp. was classified as R. tarasevichiae and probably as a new Rickettsia raoultii subspecies, and Ehrlichia spp. was classified as E. muris. The coexistence of several pathogens was found in 5.7% of tick specimens, and TBEV + Borrelia spp. was the most frequent combination.     

Autoradiographic screening for potential heterotrophs in natural algal populations of Lake Kinneret

Microautoradiography was used to screen natural phytoplankton populations from Lake Kinneret for their ability to take up radioactive organic substrates (glucose, acetic acid, amino acids, and glycollic acid. Several of the important Chlorophyta (Pediastrum spp.,Actinastrum hantzschii, Kirchncriella spp.,Coelastrum spp.,Scenedesmus spp., andTetraëdron spp.) and Cyanophyta (Microcystis spp.,Merismopedia sp.,Chroococcus spp., andAnabaena spp.) showed consistent organic uptake. However, the principal bloom former, the dinoflagellatePeridinium cinctum fawestii and most of the pelagic algae observed, never assimilated any of the above substrates. Autoradiographic surveys permit rapid screening of algal populations for species which are capable of taking up organic matter and can indicate potential facultative heterotrophs.     

First detection of anti-Besnoitia spp. specific antibodies in horses and donkeys in Italy

Among Apicomplexa protozoa infecting equids, Besnoitia spp., Toxoplasma gondii and Neospora spp. represent important issues from a sanitary and zootechnical viewpoint. However, only scarce epidemiological data are available on the spread of the infections in horses and donkeys in Europe. Therefore, a serosurvey was planned to estimate the prevalence of these Sarcocystidae species in Italian equids. Serum samples from 268 horses and 18 donkeys raised in Italy were collected and serologically analyzed to detect anti-Besnoitia spp., anti-T. gondii and anti-Neospora spp. antibodies: an approach based on an initial screening by in-house ELISA followed by a confirmatory WB was used. Two horses (0.7%) and four donkeys (22.2%), showed antibodies anti-Besnoitia spp. Ten horses (3.7%) resulted positive to T. gondii and one of these (0.4%) was seropositive also to Neospora spp. This is the first detection of anti-Besnoitia spp. specific antibodies in Italian horses and donkeys. The study confirmed the circulation of Besnoitia spp. among equids in Europe. Low prevalence of T. gondii and Neospora spp. in horses raised in Italy was reported. Nevertheless, it is noteworthy to consider that consumption of horse meat could represent a source for human toxoplasmosis.     

Surveillance and characterisation of Cronobacter spp. in Czech retail food and environmental samples

Cronobacter spp. (formerly Enterobacter sakazakii) are emerging, opportunistic pathogens that are linked with food-borne infections in neonates and infants. In the present study, 291 samples of food, 36 samples from a dairy farm and 140 samples of dust from vacuum cleaners were examined for the presence of Cronobacter spp. using chromogenic media and biochemical tests. Altogether, 72 Cronobacter spp. strains were isolated in accordance with the reference standard ?SN P ISO/TS 22964 (2006). No Cronobacter spp. strains were detected in 10 samples of infant milk formula or in samples from a dairy farm. Twelve out of 20 positive food samples were dry products. The incidence of Cronobacter spp. in instant and powdered products and spices (12 positive isolates out of 82 samples) was significantly higher than that in other foods (P?=?0.002), but lower than that in samples of dust (52 isolates; P?<?0.001). The incidence of Cronobacter spp. in dust from restaurants, bars and hotels (13 positive isolates in 20 samples) was significantly higher than that in dust from households (P?=?0.010). The polymerase chain reaction assay for the species-specific detection of the rpoB gene was performed in 49 isolates. Thirty-four Cronobacter spp. isolates were identified as Cronobacter sakazakii, nine isolates as Cronobacter malonaticus and one isolate as Cronobacter turicensis.     

The super repertoire of type IV effectors in the pangenome of Ehrlichia spp. provides insights into host-specificity and pathogenesis

The identification of bacterial effectors is essential to understand how obligatory intracellular bacteria such as Ehrlichia spp. manipulate the host cell for survival and replication. Infection of mammals–including humans–by the intracellular pathogenic bacteria Ehrlichia spp. depends largely on the injection of virulence proteins that hijack host cell processes. Several hypothetical virulence proteins have been identified in Ehrlichia spp., but one so far has been experimentally shown to translocate into host cells via the type IV secretion system. However, the current challenge is to identify most of the type IV effectors (T4Es) to fully understand their role in Ehrlichia spp. virulence and host adaptation. Here, we predict the T4E repertoires of four sequenced Ehrlichia spp. and four other Anaplasmataceae as comparative models (pathogenic Anaplasma spp. and Wolbachia endosymbiont) using previously developed S4TE 2.0 software. This analysis identified 579 predicted T4Es (228 pT4Es for Ehrlichia spp. only). The effector repertoires of Ehrlichia spp. overlapped, thereby defining a conserved core effectome of 92 predicted effectors shared by all strains. In addition, 69 species-specific T4Es were predicted with non-canonical GC% mostly in gene sparse regions of the genomes and we observed a bias in pT4Es according to host-specificity. We also identified new protein domain combinations, suggesting novel effector functions. This work presenting the predicted effector collection of Ehrlichia spp. can serve as a guide for future functional characterisation of effectors and design of alternative control strategies against these bacteria.     

Identification of Armored Scales (Hemiptera: Diaspididae) on Bamboos in Korea

This work provides an identification key to adult females of six species and four genera of armored scales including a new record, Kuwanaspis hikosani (Kuwana) that may be found on bamboos (Poaceae: Arundinaria spp., Phyllostachys spp., Pseudosasa spp., and Sasa spp.) in Korea.     

Checklist of the Braulidae,Camillidae, Diastatidae and Drosophilidae of Finland (Diptera,Ephydroidea)

A checklist of the Braulidae (1 species), Camillidae (3 spp.), Diastatidae (10 spp.) and Drosophilidae (67 spp.) of Finland is presented. Campichoetinae is included as a subfamily of Diastatidae.     

Molecular survey of canine vector-borne diseases in stray dogs in Thailand

Despite the large population of stray dogs in Thailand, there is limited information on the prevalence of canine vector-borne diseases (CVBDs). In this study, a molecular survey was conducted to determine the prevalence of Babesia spp., Ehrlichia canis, Hepatozoon spp., Anaplasma platys and Mycoplasma spp. in dogs in Thailand. Of the 181 dog blood samples tested by PCR, 78/181 (43.1%) were found to be infected with one or more pathogens. The overall prevalence rates of Mycoplasma spp., Hepatozoon spp., Babesia spp., A. platys and E. canis infections were 19.9%, 18.8%, 9.4%, 4.4% and 3.9%, respectively. To the authors' knowledge, this is the first report of Mycoplasma infection in Thailand in dogs. The current findings are important for future surveillance of CVBDs and designing appropriate approaches for diagnosis and control for the diseases in Thailand.     

Geometric morphometric study of geographic and host-related variability in Aceria spp. (Acari: Eriophyoidea) inhabiting Cirsium spp. (Asteraceae)

The russet mite, Aceria anthocoptes (Nalepa), is the only eriophyoid that has been recorded on Cirsium arvense (L.) Scop. It has been noted in several European countries and recently in the USA. In this study we explored the geographic and host-related variability of Aceria spp. inhabiting different Cirsium spp. We applied landmark-based geometric morphometric methods to study morphological variability of three body regions (ventral, coxigenital and prodorsal) of 13 Aceria spp. populations inhabiting five Cirsium spp. in Serbia (Europe) and four Cirsium spp. in Colorado (North America). Analyses of size and shape variation revealed statistically significant differences between Aceria spp. living on European native and North American native Cirsium spp., as well as between A. anthocoptes s.s. inhabiting European C. arvense and North American C. arvense. The coxigenital region was the most informative when considering inter-population shape differences. European Aceria spp. dwelling on Cirsium spp., including A. anthocoptes s.s. from C. arvense, are characterized by higher inter-population size and shape variability than their North American counterparts. This finding supports a Eurasian origin of A. anthocoptes, presumed to consist of a complex of cryptic taxa probably coevolved with host plants in the native environment. Morphological similarity among Aceria spp. inhabiting North American native Cirsium spp. may indicate that speciation of A. anthocoptes started relatively soon after the host shift to plants different from C. arvense in the invaded region.     

In vivo and in vitro management of Meloidogyne incognita (Tylenchida: Heteroderidae) using rhizosphere bacteria,Pseudomonas spp. and Serratia spp. compared with oxamyl

Biological control using rhizosphere bacteria, Pseudomonas spp. and Serratia spp. is a prospective alternative technique to overcome plant parasitic nematodes infection. So, the current study was conducted in vitro on five egg-masses, 100 free eggs and 100 infective juveniles (IJs) of Meloidogyne incognita as well as greenhouse treatments on Luffa aegyptiaca L. to evaluate the nematicidal potential of six strains belong to Pseudomonas spp. and Serratia spp. as compared to oxamyl.Results showed that the inhibitory effect and juvenile mortality varied according to bacteria species, strains and exposure time. All the tested bacteria significantly (P ≤ 0.05) inhibited egg hatching and increased juvenile mortality in vitro. After 3 days of treatment, Pseudomonas spp. were more effective against eggs (48.31to 55.15%) and IJs (20.98 to 25.30%) than Serratia spp. (44.55 to 49.75% with eggs) and (19.06 to 21.61% with IJs), respectively. In the pot experiment, Luffa aegyptiaca L. treated with Serratia spp. and Pseudomonas spp. displayed significantly higher (P ≤ 0.05) levels of growth (as indicated by root length, fresh roots weight and fresh shoots weight) compared to control plants and significantly (P ≤ 0.05) suppressed galling (number of galls) and reproduction (as indicated by number of egg-masses on roots and number of eggs and juveniles in pot soil). Meanwhile, among the treated plants, Serratia spp. and Pseudomonas spp. gave the best results in shoot weight of pots infected by eggs of M. incognita than those infected with IJs as compared with positive control. While, oxamyl treatment gave the best results in pots infected by eggs and IJs.The lowest galling (gall index), number of eggs and juveniles in soil was observed in the treatment with mixture of Serratia spp. and Pseudomonas spp. as well as, enhanced growth of sponge gourd more than application each of them alone. Pots treated with oxamyl overwhelmed those treated with mixture of Serratia spp. and Pseudomonas spp.     

Zonal distribution and seasonal vertical migration of copepod assemblages in the Scotia Sea

Large, biomass-dominant Southern Ocean copepod species have been much studied, but small and mesopelagic species also play major rôles in these ecosystems. However, little is known of some basic aspects of their ecology. To address this, the abundances of 23 copepod species and genera were analysed from 72 stations sampled during the Discovery Expeditions in the 1920s to 1950s. Stratified net samples, usually to a depth of 1000?m, provided year-round coverage in the Scotia Sea from the Subantarctic Front to the Weddell-Scotia Confluence. Small copepods (Microcalanus pygmaeus, Ctenocalanus spp., Oncaea spp. and Oithona spp.) formed ～75% of total copepod abundance in the top 1000?m across all major zones. Oithona spp. composed ～40% of copepod numbers in the Polar Front area and to its south: further north their importance declined. All mesopelagic taxa except for the warmer-water species Metridia lucens and Pleuromamma robusta, extended throughout the entire study area, with smaller regional differences than for the shallower-living species. The species showed a continuum of temperature ranges, and there was no evidence that the Polar Front was a major biogeographic boundary to their distribution. Indeed, several important species, including Oithona spp. (mainly Oithona similis), Ctenocalanus spp., Metridia lucens and Rhincalanus gigas reached maximum numbers in this area. Total copepod abundance was thus higher in the vicinity of the Polar Front than in any other region. Only two copepod families made pronounced seasonal vertical migrations: Eucalaniidae (Eucalanus longiceps and R. gigas) and Calaniidae (Neocalanus tonsus, Calanoides acutus, Calanus simillimus and Calanus propinquus). Some evidence for a winter descent was found for Ctenocalanus spp. and some deeper-living groups: Euchaeta spp. and the Metridiidae, although their migrations were not so great as for the eucalanids and calanids.     

Evaluation of Methyl Bromide Alternatives Efficacy against Soil-Borne Pathogens,Nematodes and Soil Microbial Community

Methyl bromide (MB) and other alternatives were evaluated for suppression of Fusarium spp., Phytophthora spp., and Meloidogyne spp. and their influence on soil microbial communities. Both Fusarium spp. and Phytophthora spp. were significantly reduced by the MB (30.74 mg kg^-1), methyl iodide (MI: 45.58 mg kg^-1), metham sodium (MS: 53.92 mg kg^-1) treatments. MS exhibited comparable effectiveness to MB in controlling Meloidogyne spp. and total nematodes, followed by MI at the tested rate. By contrast, sulfuryl fluoride (SF: 33.04 mg kg^-1) and chloroform (CF: 23.68 mg kg^-1) showed low efficacy in controlling Fusarium spp., Phytophthora spp., and Meloidogyne spp. MB, MI and MS significantly lowered the abundance of different microbial populations and microbial biomass in soil, whereas SF and CF had limited influence on them compared with the control. Diversity indices in Biolog studies decreased in response to fumigation, but no significant difference was found among treatments in PLFA studies. Principal component and cluster analyses of Biolog and PLFA data sets revealed that MB and MI treatments greatly influenced the soil microbial community functional and structural diversity compared with SF treatment. These results suggest that fumigants with high effectiveness in suppressing soil-borne disease could significantly influence soil microbial community.