Anaplasma infection in free-ranging Iberian red deer in the region of Castilla-La Mancha, Spain

Organisms in the genus Anaplasma are obligate intracellular pathogens that multiply in both vertebrate and invertebrate hosts. The type species, Anaplasma marginale, causes bovine anaplasmosis and infects erythrocytes of the vertebrate host and undergoes a complex developmental cycle in ticks which serve as biological vectors. Infected cattle, wild ruminants and ticks can all serve as reservoirs of A. marginale. In this study, hunter killed Iberian red deer (Cervus elaphus hispanicus) from the region of Castilla-La Mancha in southwestern Spain were tested for Anaplasma infection. We found that 10% of the deer examined were seropositive for Anaplasma. Three A. marginale strains were subsequently obtained from salivary glands of Hyalomma marginatum that were removed from these deer, and the sequence of the major surface protein (msp)4 gene was determined for each strain and used for phylogenetic studies. Maximum parsimony analyses of msp4 sequences from H. marginatum ticks in comparison with New World cattle and bison isolates reported previously, suggested different origins for these Spanish A. marginale strains. The results of this study demonstrated that Iberian red deer are naturally infected with Anaplasma, and may therefore serve as a wildlife reservoir of the pathogen. Although the link between deer infection and the strains of A. marginale identified in ticks was not established, H. marginatum and Rhipicephalus bursa were identified as potential biological vectors for A. marginale in this region and may effect transmission of A. marginale between deer and cattle populations.     

The O-antigen of Salmonella enterica serotype Enteritidis PT4: a significant factor in gastrointestinal colonisation of young but not newly hatched chicks

The lipopolysaccharide of Salmonella and other Gram negative pathogenic species has been implicated as a major virulence determinant and in this study we report the role of LPS of S. Enteritidis in the colonisation and persistent gastrointestinal infection of young poultry. The gene encoding the unique O-antigen ligase, waaL, was mutated by insertional inactivation in a well characterised S. Enteritidis strain, S1400/94. The waaL mutant, designated PCP, produced rough colonies on agar medium, did not agglutinate O9 antiserum, did not produce an LPS ladder on silver stained gels and was serum sensitive. PCP and a nalidixic acid marked derivative of S1400/94 (S1400/94 Nalr) were used to orally challenge young chicks, separately and together in competitive index experiments. At post-mortem examination of 1-day-old chicks challenged S1400/94 Nalr and PCP separately there were no significant differences in the numbers of S1400/94 Nalr and PCP bacteria in tissues sampled on days 1, 2, and 5. By day 42 after challenge S1400/94 Nalr bacteria were recovered in significantly higher numbers than PCP from the caecal contents (P < 0.001). In competitive index studies in the 1-day-old chick PCP colonised, invaded and persisted in lower numbers than S1400/94 Nalr. In 4-week-old chicks challenged separately, PCP bacteria were recovered from all tissues examined in significantly lower numbers than S1400/94 Nalr. In competitive index experiments in 4-week-old chicks, PCP was not detected at any site and at any time point. Therefore, the O-antigen of S. Enteritidis plays an important role in poultry infections although this role is less important in the newly hatched chick.     

Remote sensing and epidemiology: examples of applications for two vector-borne diseases

Remote sensing techniques have greatly contributed to improve our capacity to observe our environment and its processes. For about 15 years, the use of satellite images for epidemiological purposes has been largely promoted to determine diseases distributions and their variations through time. In some circumstances, when diseases are strongly related to environmental data such as climate, vegetation or land-use, radiation values can be included in prediction models. In other cases, remote sensing data provide information for drawing thematic layers involved in the epidemiological processes, which may differ according to the different ecotypes and ecosystems. According to its final goal, the users can choose from the panel of available radiometers with specific characteristics including spatial resolution and frequency of data. In this paper, two examples of major vector-borne diseases, namely Animal Trypanosomosis and Bluetongue, illustrate these applications.     

Development of a two-step nested duplex PCR assay for the rapid detection of Brachyspira pilosicoli and Brachyspira intermedia in chicken faeces

Avian intestinal spirochaetosis (AIS) is an infection of the caeca and/or colo-rectum of laying and meat breeder hens caused by anaerobic intestinal spirochaetes of the genus Brachyspira. AIS can result in a variety of symptoms, including delayed and/or reduced egg production, and increased faecal water content. The two most commonly reported Brachyspira species involved in AIS are Brachyspira pilosicoli and Brachyspira intermedia, and their detection and identification can be difficult and time consuming. In the current study a two-step nested duplex PCR (2S-N-D-PCR) was developed for the detection of these two species, using DNA extracted from washed chicken faeces. In the first step, a duplex PCR (D-PCR) amplifying Brachyspira genus-specific portions of the 16S rRNA and NADH oxidase (nox) genes was undertaken on the washed faeces. In the second step, a nested D-PCR was used that amplified species-specific portions of the 16S rRNA gene of B. pilosicoli and the nox gene of B. intermedia from the amplicons produced in the first step. The 2S-N-D-PCR was rapid and specific, and could be used to detect approximately 10(3) cells of each spirochaete species per gram of washed faeces. When tested on 882 chicken faecal samples from infected flocks, it detected 4-5% more positive faecal samples than did the standard method of selective anaerobic culture followed by individual species-specific PCR assays conducted on the growth on the primary plate. The application of this new technique should improve diagnostic capacity, and facilitate further studies on AIS.     

Detection of the astA (EAST1) gene in attaching and effacing Escherichia coli from ruminants

A total of 206 attaching and effacing Escherichia coli strains from ruminants were analysed for the presence of the astA (EAST1) and bfpA genes. None of these strains was bfpA-positive. The percentage of enteropathogenic E. coli (EPEC) strains astA-positive found in healthy cattle (15.6%) suggests that this animal species may be a significant reservoir of atypical EPEC potentially pathogenic for humans.     

Escherichia coli O115 forms fewer attaching and effacing lesions in the ovine colon in the presence of E. coli O157:H7

Escherichia coli O115 has been isolated from healthy sheep and was shown to be associated with attaching-effacing (AE) lesions in the large intestine. Following previous observations of interactions between E. coli O157 and O26, the aim of the present study was to assess what influence an O115 AE E. coli (AEEC) would have on E. coli O157 colonisation in vitro and in vivo. We report that E. coli O115- and O157-associated AE lesions were observed on HEp-2 cells and on the mucosa of ligated ovine spiral colon. In single strain inoculum, E. coli O115 associated intimately with HEp-2 cells and the spiral colon in greater numbers than E. coli O157:H7. However, in mixed inoculum studies, the number of E. coli O115 AE lesions was significantly reduced suggesting negative interference by E. coli O157. Use of the ligated colon model in the present work has allowed in vitro observations to be extended and confirmed whilst using a minimum of experimental animals. The findings support a hypothesis that some AEEC can inhibit adhesion of other AEEC in vivo. The mechanisms involved may prove to be of utility in the control of AE pathovars.     

Spatial and temporal patterns at small scale in Austrocedrus chilensis diseased forests and their effect on disease progression

Austrocedrus chilensis is an endemic conifer of Patagonia that suffers a widespread mortality whose causes are a topic of discussion. Since Phytophthora austrocedrae is the most probable cause, we proposed that the spatial and temporal patterns of disease at small scale should reflect pathogen behavior. We aimed at characterizing the spatial and temporal patterns of diseased trees in different soil types and the effect of microsite variability on diseased trees spatial pattern. The spatial pattern of disease was influenced by soil type and tree density. In clay soils with low disease incidence (ca. 25%), the spatial pattern was random and not influenced by abiotic microsite conditions. When disease incidence increased (ca. 70%), concurring with denser plots, the spatial pattern was clustered, as a result of an infection process, and it was independent of microsite variability. In soils with better drainage conditions, that is, alluvial soils with volcanic ash input and coarse textured volcanic soils, the disease was clustered and associated with flat microtopographies. The progression of the disease at small scale was influenced by soil, precipitation and tree density. The spatial and temporal patterns of disease progression were associated with a contagion process and with environmental variables that affect drainage, coinciding with Phytophthora biology and requirements. Our results concur in pointing at Phytophthora as the cause of A. chilensis disease in the study area. Management practices should be urgently applied in order to minimize the spread of the inoculum.     

Lead Contamination in Feed Affects Performance, Egg Quality and Antioxidant Capacity of Laying Hens

本试验旨在研究不同剂量铅对蛋鸡生产性能、蛋品质以及血清、肝脏和肾脏中与抗氧化性能相关指标的影响.选用40周龄生产性能相近的海兰褐蛋鸡576羽,随机分为4组:对照组、试验1组(T1组)、试验2组(T2组)、试验3组(T3组),每组4个重复,每个重复36羽.对照组饲喂基础饲粮,T1、T2、T3组分别饲喂在基础饲粮中添加15、30、60 mg/kg铅的试验饲粮.试验期8周.结果表明:1)各组间产蛋率和料蛋比均无显著差异(P＞0.05),1～8周T3组平均蛋重显著低于对照组(P＜0.05),1～8周T1、T2、T3组平均日采食量分别较对照组降低了3.72％ (P ＜0.05)、1.93％ (P ＞0.05)、3.63％ (P ＜0.05).2)与对照组相比,试验组蛋白高度和哈夫单位均有降低趋势,除T3组蛋白高度(P＜0.05)外均差异不显著(P＞0.05);T3组蛋壳强度和蛋壳厚度最低,第4周和第8周时T3组蛋壳强度分别比对照组降低了21.89％、16.84％(P＜0.05);蛋壳厚度的变化趋势与蛋壳强度一致,均随着铅添加量的增加而降低,呈现一定的剂量-效应关系.3)与对照组相比,各试验组血清、肝脏和肾脏中谷胱甘肽过氧化物酶活性均显著降低(P＜0.05);丙二醛(MDA)含量均在一定程度上升高,T3组血清和肝脏中MDA含量均较对照组显著上升(P＜0.05),各试验组肾脏中MDA含量均显著高于对照组(P＜0.05);血清、肝脏和肾脏中还原型谷胱甘肽含量和总抗氧化能力以及总超氧化物歧化酶活性均呈现降低趋势.综上所述,饲料中铅污染可以导致蛋品质降低,诱导脂质过氧化作用,降低蛋鸡抗氧化能力.