两株blaNDM-5基因介导的碳青霉烯耐药禽源大肠杆菌ST10和ST354耐药性

【背景】近年来,对碳青霉烯类抗生素耐药的肠杆科细菌的出现对公众的健康构成了严重的威胁。【目的】为了解家禽养殖中肠道菌对碳青霉烯类抗生素耐药情况,对陕西西安和浙江海宁两地的鸡和鸭养殖场粪便进行耐药菌筛选并分析。【方法】在含美罗培南的LB琼脂平板上筛选可疑菌落并利用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)鉴定。通过微量肉汤稀释法进行最小抑菌浓度(minimum inhibitory concentration,MIC)测定。通过Illumina HiSeq平台进行全基因组测序。使用Res Finder数据库预测获得性耐药基因。利用S1酶切后脉冲场电泳(pulsed-field gel electrophoresis,PFGE)和Southern blot杂交进行质粒和碳青霉烯类耐药基因的确认。【结果】分离出两株碳青霉烯类耐药的大肠杆菌HN1-26和XN3-1,均对头孢他啶、头孢噻呋、氨苄西林、奥格门丁、复方新诺明、磺胺异恶唑、恩诺沙星、氧氟沙星、美罗培南、四环素耐药。此外,XN3-1对氟苯尼考和大观霉素耐药。菌株HN1-26和XN3-1的多位点序列分型(multilocus sequence typing,MLST)分别为ST354和ST10。两株菌对美罗培南的MIC值均为64μg/m L,但是在含美罗培南的LB平板上,菌株HN1-26比菌株XN3-1生长得更快。两株菌都存在携带bla_(NDM-5)基因的Inc X3型质粒。两个质粒的序列完全一致,大小为46 161 bp,包含接合转移元件,能够以大肠杆菌作为受体细胞进行转移。【结论】Inc X3型质粒是转移bla_(NDM-5)基因的重要载体,在我国畜禽养殖中具有扩散风险。     

Consequences of the Ban of By-Products from Terrestrial Animals in Livestock Feeding in Germany and The European Union: Alternatives,Nutrient and Energy Cycles,Plant Production,and Economic Aspects

Consequences of the ban of meat and bone meal (MBM) and animal fat with regard to livestock feeding, cropping, ecology and economy where investigated with an inter-disciplinary approach for Germany and the European Union. Calculations were made for different production systems with pigs and poultry on the basis of statistical data for the production and for the feed markets as well as from requirement data for the respective species and production system. (1.) The ban of MBM from feeding caused a need for alternative protein sources. If all the amount of protein from MBM is to be replaced by soybean meal, in Germany and the EU about 0.30 and 2.30 · 10⁶t would be needed each year (supplementary amino acids not considered). Alternatively, doubling the grain legume acreage in Germany to about 420,000ha would supply a similar amount of protein. A wider application of phase feeding with adjusted dietary amino acid concentrations, however, would allow for saving protein to an extent which is similar to the amount of protein that was contributed by MBM in recent years. Thus, the ban is a minor problem in terms of ensuring amino acid supply. (2.) However, alternative plant ingredients cannot compensate for the gap in P supply that is caused by the ban. An additional demand for inorganic feed phosphates of about 14,000 and 110,000t per year is given in Germany and the EU, respectively. So far, this gap is filled almost completely by increased mining of rock phosphates. Alternatively, a general application of microbial phytase to all diets would largely fill this gap. Until the ban, MBM contributed to 57% of the supplementation of P that was needed for pigs and poultry. The ban of MBM makes large amounts of P irreversibly disappearing from the food chain. (3.) Energy from slaughter offal and cadavers can be utilized in different technologies, in the course of which the efficiency of energy utilisation depends on the technology applied. It is efficient in the cement work or rotation furnace if heat is the main energy required. In contrast, the energetic efficiency of fermentation is low. (4.) Incineration or co-incineration of MBM and other by-products causes pollution gas emissions amounting to about 1.4kg CO₂ and 0.2kg NO_x per kg. The CO₂ production as such is hardly disadvantageous, because heat and electrical energy can be generated by the combustion process. The prevention of dangerous gaseous emissions from MBM burning is current standard in the incineration plants in Germany and does not affect the environment inadmissibly. (5.) The effects of the MBM ban on the price for compound feed is not very significant. Obviously, substitution possibilities between different feed ingredients helped to exchange MBM without large price distortions. However, with each kg MBM not used in pig and poultry feeding economic losses of about 0.14¢ have to considered. In conclusion, the by far highest proportion of raw materials for MBM comes as by-products from the slaughter process. Coming this way, and assuring that further treatment is safe from the hygienic point of view, MBM and animal fat can be regarded as valuable sources of amino acids, minerals and energy in feeding pigs and poultry. Using them as feedstuffs could considerably contribute to the goal of keeping limited nutrients, phosphorus in particular, within the nutrient cycle and dealing responsible with limited resources.     

Microbial challenges of poultry meat production

Food safety and shelf-life are both important microbial concerns in relation to broiler meat production. Focus is mainly placed on the absence or control of potentially pathogenic microbes such as Salmonella spp. and Campylobacter spp. but, from the commercial point of view, other spoilage bacteria also play a role as potential threats. Regarding food safety, the primary target should be the production of pathogen-free live animals, thus allowing slaughter plants to keep the processing line free of those microorganisms.Consumers believe that quality of foods from organic production is superior to foods from conventional production. The aim of the present study was to evaluate and compare the bacterial quality of chicken meat from organic and conventional production on the basis of traditional meat quality criteria. Fresh free grazing broiler carcasses were purchased directly from rural households (n = 80) and fresh retail chicken parts from conventional broiler carcasses from the local supermarkets in the region of Epirus (Poultry Producers Association. Arta) (n = 200).The samples were microbiologically tested for the presence of bacteria such as: Salmonella spp., Listeria monocytogenes, Staphylococcus aureus, Enterobacteriaceae, Escherichia coli, Campylobacter spp., and C. perfringens. Total count of aerobic mesophilic bacteria was also determined. Bacteriological tests were performed by means of standard methods of isolation and identification of individual species of bacteria according to ISO requirements. API-tests (bioMerieux) and Vitek 2 Identification System (bioMerieux) were used for biochemical determination. High levels of microbial contamination and occurrence of pathogenic bacteria at then fresh free grazing broiler carcasses reflect the poor hygienic quality of the slaughter conditions in the rural households.     

Differential Toll-Like Receptor (TLR) mRNA Expression Patterns during Chicken Embryological Development

Toll-like receptors (TLRs), important components of innate immune response, play a pivotal role in early recognition of pathogen as well as in the initiation of robust and specific adaptive immune response. In the present study, the expression profile of chicken TLRs (TLR2A, TLR3, TLR4, TLR5, TLR7, TLR15, and TLR21) in various chicken embryonic tissues during embryo development was examined by real-time PCR assay. All the TLR mRNAs were expressed in whole embryonic tissue as early as 3rd embryonic day (ED). Four of the seven TLRs (TLR2, TLR3, TLR4, and TLR7) mRNA expressions were significantly (P < 0.01) higher at 12ED relative to expression at 3 ED, whereas TLR15 mRNA expression was significantly (P < 0.01) higher on 7ED and TLR5 and 21 were highly expressed on 18 ED. Among all the TLRs investigated TLR4 mRNA was the highest expressed and TLR15 mRNA expression was the lowest in all tissues during chicken embryo development. Tissue wise analysis of mRNA expression of TLRs showed that liver expressed significantly (P < 0.01) higher levels of most of the genes (TLR2, TLR4, and TLR21). However no significant difference was found in TLR15 mRNA expression among the tissues during development. Our results suggest the innate preparedness of chicken embryos and also a possible role for TLRs in the regulation of chicken embryo development that needs to be further evaluated.     

Efficacy of Trichogramma evanescens in controlling the grape berry moth Lobesia botrana in grape farms in Egypt

Abstract

The present work was conducted to evaluate the efficacy of Trichogramma evanescens (Westwood) in controlling the grape berry moth Lobesia botrana (Schiff) in two grape farms, in El-Beheira and El-Gharbia Governorates, northern Egypt during 2004 and 2005 seasons. T. evanescens was mass produced on Sitotroga cerealella (Olivier) eggs in the National Research Centre in Egypt. The horizontal and vertical searching activity of T. evanescens was studied to determine the proper way of distributing the Trichogramma cards in vineyards. Field experiments were also conducted to evaluate T. evanescens efficacy to control L. botrana on a large scale. Parasitism by T. evanescens on L. botrana eggs was greatly affected with the horizontal or vertical distance from the release points as well as with the rate of release. Parasitism reached over 97% and the percentage of reduction in damage caused by the pest reached 96.8% in treated plots. A significant increase in the crop was achieved in treated plots. T. evanescens could be a potential candidate for biological control of the grape moth in vineyards.     

Use of enzymes to improve nutrient availability in poultry feedstuffs

Enzyme supplementation of cereal-based diets can significantly improve chick performance by increasing the rate of gain, efficiency of feed utilization, the apparent metabolizable energy and digestibilities of dry matter, fat and protein, with excellent improvements being obtained with diets containing rye, oats and barley. Less dramatic results are often obtained with wheat. In addition, enzyme treatment decreases the moisture content of excreta, which, together with improved dry matter digestibility, reduces the total amount of excreta produced and therefore reduces management and environmental problems. Enzyme supplementation also improves the nutritional value of lupins, and reduces the length and size of various sections of the gastrointestinal tract and the size of the pancreas of chickens. Dose-response studies with different amounts of supplemental enzymes when added to a rye-based diet demonstrated that there was a high linear correlation (r²>0.91, P < 0.05) between the concentration of enzyme when transformed into its logarithmic values and the corresponding improvements in weight gain or the feed-to-gain ratio. The log-linear model shows that for every ten-fold increase in the amount of enzyme in the diet there was a two-fold and not a ten-fold incremental improvement in chick performance. These studies suggest that there is a simple relationship between the amount of enzyme added to the diet and the resulting improvements that are obtained. Overall, enzymes when properly used can produce significant improvements in chick performance and can reduce the excretion of undigested nutrients.     

Quantification of thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> spp. in broilers during meat processing

Campylobacter spp. is a common cause of gastrointestinal illness. Since animal products, especially poultry meat, are an important source of human outbreaks of campylobacteriosis, tracing back to processing and initial production is of great interest. Samples were collected at a German poultry slaughterhouse for the estimation of the prevalence of Campylobacter at different processing steps. Quantification of Campylobacter in each of the samples was also performed. Out of 99 samples examined, 51 (51.5%) were positive for Campylobacter, with bacterial counts ranging from log(10) 6.5 cfu sample(-1) for carcasses to log 3.6 cfu ml(-1) for scalding water. The Campylobacter isolates (n = 51) were subtyped by pulsed-field gel electrophoresis using SmaI and KpnI restriction enzymes. Molecular typing showed a multitude of strains with different molecular patterns. Strains found in cloacal swabs before processing could also be isolated from carcasses at different processing steps.     

Wind Energy Development and Wildlife Conservation: Challenges and Opportunities

ABSTRACT Wind energy development represents significant challenges and opportunities in contemporary wildlife management. Such challenges include the large size and extensive placement of turbines that may represent potential hazards to birds and bats. However, the associated infrastructure required to support an array of turbines—such as roads and transmission lines—represents an even larger potential threat to wildlife than the turbines themselves because such infrastructure can result in extensive habitat fragmentation and can provide avenues for invasion by exotic species. There are numerous conceptual research opportunities that pertain to issues such as identifying the best and worst placement of sites for turbines that will minimize impacts on birds and bats. Unfortunately, to date very little research of this type has appeared in the peer-reviewed scientific literature; much of it exists in the form of unpublished reports and other forms of gray literature. In this paper, we summarize what is known about the potential impacts of wind farms on wildlife and identify a 3-part hierarchical approach to use the scientific method to assess these impacts. The Lower Gulf Coast (LGC) of Texas, USA, is a region currently identified as having a potentially negative impact on migratory birds and bats, with respect to wind farm development. This area is also a region of vast importance to wildlife from the standpoint of native diversity, nature tourism, and opportunities for recreational hunting. We thus use some of the emergent issues related to wind farm development in the LGC—such as siting turbines on cropland sites as opposed to on native rangelands—to illustrate the kinds of challenges and opportunities that wildlife managers must face as we balance our demand for sustainable energy with the need to conserve and sustain bird migration routes and corridors, native vertebrates, and the habitats that support them.     

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0–41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.