Phytotoxic Effects of Antibiotics on Terrestrial Crop Plants and Wild Plants: A Systematic Review.

This review examines the state of knowledge on the phytotoxic effects of antibiotics on terrestrial crop plants and wild (non-crop) plants with the goal of evaluating differences in their sensitivity. This is important because environmental risk assessments of antibiotics currently consider their potential effects only on crop species but not wild species. Overall, we analysed 275 datasets consisting of antibiotic-plant species-endpoint combinations for germination (mg/L) and 169 datasets for plant growth (elongation and biomass) (mg/kg). EC10 and EC50 of each parameter were compared using a quotient approach, in which the geometric mean and the 5th percentile of the crop data were divided by wild data. Quotients were > 1 for elongation growth, suggesting that wild species were more sensitive than crops, while they were < 1 for biomass growth, suggesting quite the contrary. However, < 1% of the data in each dataset came from wild species, preventing definitive conclusions. Merging crop and wild data to evaluate differences in sensitivity among classes of antibiotics and plant families, we found using a linear mixed effect model and post hoc test that plants were most sensitive to phenicol and least sensitive to macrolides and tetracyclines. Further work must be conducted to gain a better understanding of the phytotoxic effects of antibiotics on terrestrial wild plants and subsequently assess whether the current approach to environmental risk assessment of antibiotics is sufficient to protect plant biodiversity.

Assessing the benefits of composting poultry manure in reducing antimicrobial residues, pathogenic bacteria, and antimicrobial resistance genes: a field-scale study.

The poultry industry in the European Union produces 13 million tons of manure annually, which represents a major health and environmental challenge. Composting is an environmental-friendly technique for the management of manure, but there are few studies about antibiotic residues and antimicrobial resistances at a field scale. The goal of this study was to determine if the composting of poultry manure at a field scale would result in the reduction of antibiotic residues, pathogenic bacteria, and antibiotic resistance genes (ARGs) in the final fertilizer product. A 10-week composting of poultry manure spiked with enrofloxacin, doxycycline, and ciprofloxacin was performed. The determination of antibiotics residues and 22 selected ARGs was carried out together with the identification of bacteria by metagenomics. In the case of ciprofloxacin and doxycycline, a 90% decrease was observed after composting for 3 weeks. Sixteen ARGs were detected at the beginning of the experiment; 12 of them decreased from week 0 to week 10 (reduction of 73.7-99.99%). The presence of potentially pathogenic bacteria, such as, Campylobacter coli or commensal bacteria such as Escherichia coli decreases along the composting process. In conclusion, 10-week composting of poultry manure promotes the reduction of antibiotic residues and most of the ARGs and pathogenic bacteria.

Novel and highly sensitive SYBR® Green real-time pcr for poxvirus detection in odontocete cetaceans.

Poxviruses are emerging pathogens in cetaceans, temporarily named 'Cetaceanpoxvirus' (CePV, family Poxviridae), classified into two main lineages: CePV-1 in odontocetes and CePV-2 in mysticetes. Only a few studies performed the molecular detection of CePVs, based on DNA-polymerase gene and/or DNA-topoisomerase I gene amplification. Herein we describe a new real-time PCR assay based on SYBR® Green and a new primer set to detect a 150 bp fragment of CePV DNA-polymerase gene, also effective for conventional PCR detection. The novel real-time PCR was able to detect 5 up to 5 × 106 copies per reaction of a cloned positive control. Both novel PCR methods were 1000 to 100,000-fold more sensitive than those previously described in the literature. Samples of characteristic poxvirus skin lesions ('tattoo') from one Risso's dolphin (Grampus griseus), two striped dolphins (Stenella coeruleoalba) and two Guiana dolphins (Sotalia guianensis) were all positive to both our novel real time- and conventional PCR methods, even though three of these animals (a Risso's dolphin, a striped dolphin, and a Guiana dolphin) were previously negative to the conventional PCRs previously available. To our knowledge, this is the first real-time PCR detection method for Cetaceanpoxvirus, a much more sensitive tool for the detection of CePV-1 infections.

No pain, no gain: Male plasticity in burrow digging according to female rejection in a sand-dwelling wolf spider.

Behavioral plasticity allows individuals to reversibly respond to short-term variations in their ecological and social environment in order to maximize their fitness. Allocosa senex is a burrow-digging spider that inhabits the sandy coasts of South America. This species shows a reversal in typical sex roles expected in spiders: females are wanderers that visit males at their burrows and initiate courtship. They prefer males with long burrows for mating, and males prefer virgin over mated females. We tested whether female sexual rejection induced males to enlarge their burrows and if female reproductive status affected males' responses. We exposed males who had constructed burrows to: a) virgin females or b) mated females, (n=16 for each category). If female rejection occurred, we repeated the trial 48h later with the same female. As control, we maintained a group of males without female exposure (unexposed group, n=32). Rejected males enlarged their burrows more frequently and burrows were longer compared to unexposed males. However, frequency and length of enlargement did not differ according to female reproductive status. Males of A. senex showed plasticity in digging behavior in response to the availability of females, as a way to maximize the possibilities of future mating.

Importance of Ecological Factors and Colony Handling for Optimizing Health Status of Apiaries in Mediterranean Ecosystems.

We analyzed six apiaries in several natural environments with a Mediterranean ecosystem in Madrid, central Spain, in order to understand how landscape and management characteristics may influence apiary health and bee production in the long term. We focused on five criteria (habitat quality, landscape heterogeneity, climate, management and health), as well as 30 subcriteria, and we used the analytic hierarchy process (AHP) to rank them according to relevance. Habitat quality proved to have the highest relevance, followed by beehive management. Within habitat quality, the following subcriteria proved to be most relevant: orographic diversity, elevation range and important plant species located 1.5 km from the apiary. The most important subcriteria under beehive management were honey production, movement of the apiary to a location with a higher altitude and wax renewal. Temperature was the most important subcriterion under climate, while pathogen and Varroa loads were the most significant under health. Two of the six apiaries showed the best values in the AHP analysis and showed annual honey production of 70 and 28 kg/colony. This high productivity was due primarily to high elevation range and high orographic diversity, which favored high habitat quality. In addition, one of these apiaries showed the best value for beehive management, while the other showed the best value for health, reflected in the low pathogen load and low average number of viruses. These results highlight the importance of environmental factors and good sanitary practices to maximize apiary health and honey productivity.

Diagnosis of Cetacean morbillivirus: A sensitive one step real time RT fast-PCR method based on SYBR(®) Green.

Cetacean morbillivirus (CeMV) (family Paramyxoviridae, genus Morbillivirus) is considered the most pathogenic virus of cetaceans. It was first implicated in the bottlenose dolphin (Tursiops truncatus) mass stranding episode along the Northwestern Atlantic coast in the late 1980s, and in several more recent worldwide epizootics in different Odontoceti species. This study describes a new one step real-time reverse transcription fast polymerase chain reaction (real-time RT-fast PCR) method based on SYBR(®) Green to detect a fragment of the CeMV fusion protein gene. This primer set also works for conventional RT-PCR diagnosis. This method detected and identified all three well-characterized strains of CeMV: porpoise morbillivirus (PMV), dolphin morbillivirus (DMV) and pilot whale morbillivirus (PWMV). Relative sensitivity was measured by comparing the results obtained from 10-fold dilution series of PMV and DMV positive controls and a PWMV field sample, to those obtained by the previously described conventional phosphoprotein gene based RT-PCR method. Both the conventional and real-time RT-PCR methods involving the fusion protein gene were 100- to 1000-fold more sensitive than the previously described conventional RT-PCR method.

An approach for mapping the vulnerability of European Union soils to antibiotic contamination.

Release of antibiotics into the environment through the agricultural reuse of animal manure is considered a cause of chronic environmental exposure that often leads to adverse ecotoxicological effects, as well as to the introduction of antibiotic-resistant bacteria into the environment. The vulnerability of soil to antibiotic contamination plays a major role in determining the extent of the contamination and the likelihood of the emergence of antibiotic resistance and the appearance of ecotoxicological effects. It depends on soil characteristics, which vary across Europe, and antibiotic characteristics, which vary across drug classes. Understanding how soil vulnerability varies geographically for different veterinary medicinal products would be very useful for resource allocation among surveillance programmes. This paper performs risk analysis of the EU region for 12 antimicrobials using a spatial assessment performed in four steps. First, antibiotic release was estimated based on livestock density. Then exposure was estimated based on antimicrobial soil contamination. Third, consequences were modelled based on soil uses. Finally, risk was estimated by combining release, exposure and consequences using spatial multicriteria decision analysis. A final risk value for soil vulnerability was calculated for each antibiotic studied and displayed in chloropletic maps (ArcGIS 9.3). Furthermore, the Getis-Ord Gi statistic was used to identify clusters of areas at high risk for antibiotic soil contamination. Enrofloxacin was the highest-risk antibiotic in the European Union, followed by tetracyclines, tylosin and sulfodiazine. The highest risk values were found in Belgium, Ireland, Netherlands, Switzerland, Denmark, Germany and the UK. The results suggest that this methodology can be used successfully for evaluating the contamination potential of antibiotics over large areas with limited input data. This work is a preliminary step towards prioritising the use of veterinary medicinal products (VMPs), orientating monitoring studies and antimicrobial surveillance programmes, and informing sustainable decision-making for interventions designed to mitigate the risk of VMPs.

Organochlorine residues in the blubber and liver of bottlenose dolphins (Tursiops truncatus) stranded in the Canary Islands, North Atlantic Ocean.

Polychlorinated biphenyls (PCBs) and chlorinated pesticides: dichlorodiphenyltrichloroethane and its metabolites (DDTs), chlordanes (CHLs), dieldrin, and hexaclorobenzene (HCB) were detected in the blubber and liver of 11 bottlenose dolphins (Tursiops truncatus) from the Canary Islands (North Atlantic Ocean). Samples were obtained from stranded dolphins over the period 1997-2005. Among the organochlorines analyzed, PCBs and DDTs were predominant in the two tissues, followed in decreasing order by chlordane, trans-nonachlor > cis-nonachlor > dieldrin and HCB. The sum 11 PCBs in the blubber ranged between 301 and 33,212 ng g(-1) ww (990 and 136,679 ng g(-1) lw). Highly chlorinated PCBs such us CB153, CB180, and CB138 were the prominent congeners, accounting for 51% of the total PCBs. The sum DDT concentration in the blubber ranged between 147 and 21,050 ng g(-1) ww. (490-105,250 ng g(-1) lw) The main DDT metabolite was p,p'-DDE, representing 83% of DDTs in the blubber. In general, the levels of PCBs and DDTs detected were similar to those found in bottlenose dolphins in the North of Europe. The 2,3,7,8-TCDD toxic equivalent (TEQ) in blubber and liver was calculated for the toxicity assessment of mono-ortho substituted PCBs congeners (CB105, CB118, CB156). It is important to mention that TEQ values and p,p'-DDE concentration in adult male specimens are approaching the levels associated with adverse effects found in marine mammals. The information provided represents the first tissue loads of organochlorine compounds in small cetaceans from this area.

Plasma vitellogenin levels and gonadal morphology of wild carp (Cyprinus carpio L.) in a receiving rivers downstream of sewage treatment plants.

A field study was carried out in order to assess, in a wild cyprinid fish, the biological effects related to the exposure to estrogenic compounds. The gonadosomic index (GSI), plasma vitellogenin (VTG) level and histological observations of gonads were performed to evaluate the reproductive health in common carp (Cyprinus carpio) from three rivers receiving sewage treatment plant (STP) effluents (Guadarrama, Henares and Jarama rivers, Spain) and from a reference site (Lozoya river). No significative differences in male GSI between sites were described. Only in the 18% of fish from Guadarrama river were elevated plasma VTG levels detected. The presence of oocytes in the testis was not found in any of males examined. The results suggest that the released estrogenic compounds by STPs effluents are not enough capable to adversely affect the gonadal development of the fish studied. Additional studies will be required to assess the risk of estrogenic compounds on fish from other sites of this area.

A survey of potential stressor-induced physiological changes in carp (Cyprinus carpio) and barbel (Barbus bocagei) along the Tajo River.

The objective of this study was to evaluate fish response to acute stress induced by confinement after capture. Because of the previously reported presence of chemical compounds in the Tajo River basin where the study samplinig took place, an exposure biomarker to organic chemicals (retinol) was used. Cortisol and glucose were used as stress biomarkers. Plasma levels of cholesterol were used as lipidic metabolism indicators, and retinol level was used as a specific exposure biomarker. A reference site was established along 300 km of the Tajo River, and nine sampling sites were selected on the basis of whether various human activities and hydrographic characteristics were present. A total of 55 carp (Cyprinus carpio) and 52 barbel (Barbus bocagei) were examined. Cortisol and glucose levels were considered acceptable indicators of the response of the fish to induced stress. In the barbel, plasma retinol levels decreased at two of the sampling sites indicating possible exposure to organic compounds. The overall evaluation of these parameters enabled us to identify three sampling sites at which more studies should be carried out. The possible relationship between the health state of wild fish and the presence of organic compounds or sources of pollution was considered.

Identification of organic compounds and ecotoxicological assessment of sewage treatment plants (STP) effluents.

An integrated approach combining chemistry and biological methods was conducted to assess the toxicity of seven sewage treatment plant effluents. Solid phase concentration procedures were applied to facilitate the study of organic micro pollutants. A chemical analysis was performed by GC/MS. Organic fraction toxicity was determined by using bioassays such as Daphnia magna and Chlorella vulgaris tests and sub-lethal effects were also evaluated by using Salmonella typhimurium Test (mutagenicity), recombinant yeast screen (estrogenicity), and Oryzias latipes embryo-larval test. More than 49 compounds were detected in the organic fraction due to the various inputs of each effluents. The most frequently detected compounds in the effluents were bisphenol A (BPA), octylphenol (OP), 1,2-benzenedicarboxylic acid, bis(2-ethylhexyl) ester (DEHP) and 1,2-benzenedicarboxylic acid, bis(methylpropyl) ester (DBP). Biological assays showed toxicity effects on D. magna tests in all samples, whereas toxicity on C. vulgaris or S. typhimurium tests were not observed. Estrogenicity and teratogenicity were observed in several samples. The cause-effect relationship could not be established given the high chemical complexity of the effluents and the lack of information available on 70% of the detected compounds subsequent to reviewing various data bases. Nevertheless, due to the high chemical variability revealed by STP effluents, bioassay sets may provide a very useful amount of information for detecting potential toxicity risks.