Design and evaluation of primer pairs for efficient detection of avian rotavirus

The use of molecular methods for rotavirus characterisation provides increased sensitivity for typing and allows the identification of putative reassortant strains. Reagents and methods for genotyping the virus need constant modification because of the reassortant nature of the virus. This study was aimed at designing and evaluating new oligonucleotide degenerate primer pairs that provide increased sensitivity and specificity for detecting avian rotavirus. Gene-specific primer pairs were designed by analysing different rotavirus strains isolated during the last decade by downloading them from the GenBank. The alignments were generated using clustal analysis from the BioEdit program. Degenerate nucleotides were included due to the reassortant nature of rotavirus. The consensus sequences were aligned using the BioEdit program and then treated with the Fast PCR software to derive the primers. The derived primer sequences were submitted for a BLAST search to ensure alignment was exclusive to the desired target genes. The designed primers had specific bands and were efficient in detecting rotavirus in faecal samples than previously published primers. Thus, a successful surveillance of rotaviruses requires that primer pairs be updated regularly in order to detect the emergence of novel or “unusual types”, which have occurred by genetic drift causing nucleotide changes at the primer binding sites that result in typing failures. We recommend the use of the proposed primers in molecular surveillance studies for efficient detection of avian rotavirus.     

La Sota vaccination may not protect against virus shedding and the lesions of velogenic Newcastle disease in commercial turkeys

The aim of this project is to study the clinical signs and lesion of velogenic Newcastle disease (vND) in commercial turkeys, and also to find out if La Sota vaccination offered protection against these signs and lesions. The cockerels were included as positive controls. One hundred and twenty turkey poults and cockerels were divided into eight groups as follows: unvaccinated unchallenged turkeys (UUT), unvaccinated challenged turkeys (UCT), vaccinated unchallenged turkeys (VUT), vaccinated challenged turkeys (VCT), and along the same lines, the cockerel groups were UUC, UCC, VUC and vaccinated challenged cockerels (VCC). Vaccination was at 3 weeks of age while challenge was at 6 weeks of age. The unvaccinated turkeys and cockerels (UCT and UCC) showed different degrees of depression, diarrhoea and later paralysis at challenge. Total mortality was 100% in cockerels within 6 days, but 60% in turkeys. Similar but milder clinical signs were found in the VCC with a total mortality of 13.3%. The VCT showed mild drop in feed and water consumption, and no mortality. All the challenged groups had significant (p < 0.05) loss of weight when compared with their controls. Necropsy showed that while the UCC had severe proventricular haemorrhages, intestinal and caecal tonsil ulcers, the UCT had no digestive tract lesion. There was severe atrophy of the lymphoid organs in all the challenged groups. Histopathological sections of the bursa, spleen and thymus in all the challenged groups with special emphasis on the vaccinated and unvaccinated turkeys with mortalities of 0 and 60%, respectively, had very severe necrosis and depletion of the lymphoid tissue. Virus was isolated from the cloacal swabs. The haemagglutination inhibition antibodies were significantly higher (p < 0.05) in the challenged groups than the unchallenged. The above observations in the intestinal tracts of UCT are of diagnostic significance while the gross and microscopic lesions in the UCT and VCT show that La Sota vaccination may not protect turkeys against the destruction of the lymphoid organs by vND as earlier reported in chickens. This may lead to immunosuppression and production problems in areas where vND is enzootic.     

Long-term study (2005–2010) on the vaccination with BoHV-1 glycoprotein E-deleted marker vaccine in selected two dairy herds in Turkey

A follow-up study from 2005 to 2010 was carried out in two herds where eradication programme for the bovine herpes virus-1 (BoHV-1) infection depends on the vaccination with inactivated glycoprotein E-deleted vaccine that was started in 2001 following the vaccination with inactivated conventional vaccine between 1999 and 2001. For serological screening, a total of 12,976 sera sampled over several sampling times approximately 6 months of interval during 5 years (2005–2010) were tested for glycoprotein E (gE)- and glycoprotein B-specific antibodies using ELISA. According to the serological evidence, the long-term persistence of BoHV-1 antibodies, success of marker vaccine, first vaccination time of the calves in herds regularly vaccinated, etc. were discussed in this paper. In conclusion, the vaccination programme using gE (?) marker vaccines, with making efforts to prevent the other factors about transmission of infection, was suggested for the eradication of BoHV-1 infection in Turkey as many EU countries. This is the first report on the BoHV-1 eradication programme in some dairy cattle in Turkey.     

<Emphasis Type="SmallCaps">d</Emphasis>-Cloprostenol enhances estrus synchronization in tropical hair sheep

To compare the effects of PGF2α (dinoprost tromethamine) and d-cloprostenol in a two-dose protocol for estrus synchronization in hair sheep during breeding season in Yucatán, México, two experiments were conducted. In experiment 1, 61 cyclic hair sheep were divided into two groups: G1 (control n?=?30), two doses of 50 μg of dinoprost tromethamine IM with 12 days between applications, and G2 (n?=?31), two doses of 50 μg of d-cloprostenol IM at the same time interval. For determination of progesterone levels, 16 ewes from each group were randomly selected. In experiment 2, 70 cyclic hair sheep were assigned at the same treatments (G1 and G2, n?=?35) and 48 h after the second application, the ewes in estrus were detected by two vasectomized rams. Sheep with detected estrus were inseminated, and 45 days after, pregnant animals were identified by ultrasonography. An exact Fisher’s test was performed for the analysis of ewes in estrus (experiments 1 and 2) and number of pregnant ewes (experiment 2); for the comparison of time between end of treatment-estrus presentation, a survival analysis was used. Duration of estrus in hours was analyzed using a generalized mixed model (GLM) ANOVA whereas plasma progesterone concentrations were analyzed by non-linear regression. There were significant differences (P?<?0.05) in the proportion of ewes in estrus upon treatments (G1, 57% vs G2, 87% and G1, 37.1% vs G2, 65.7% in experiments 1 and 2, respectively), and between the end of treatment-onset estrus interval (P?<?0.01), survival curves showed the highest number of sheep in estrus between 40 and 48 h (G1, 43.7?+?8.05 h vs G2, 42.9?+?6.7 h, experiment 1). There were no significant differences (P?>?0.05) in duration of estrus (G1, 42?+?6.1 h, vs G2, 41.1?+?11.2 h, experiment 1) and pregnancy in the ewes that presented estrus, and were inseminated (G1, 38.4% vs 52.1%, experiment 2). With regard to concentrations of progesterone, significant differences (P?<?0.01) were found between treatments, and progesterone levels before the second application of d-cloprostenol were higher. In consideration of the results, it can be concluded that in a two-dose protocol of a luteolytic agent, more ewes presented estrus in response to d-cloprostenol compared to dinoprost tromethamine with similar pregnancy rates.     

Effects of protein-energetic supplementation frequency on growth performance and nutritional characteristics of grazing beef cattle

Two experiments were conducted to evaluate reduced supplementation frequencies for grazing beef cattle in rainy season. In experiment 1, evaluating the nutritional parameters, four rumen-cannulated Nellore bulls (BW = 410 kg) were used. In experiment 2, evaluating animal performance, 48 Nellore bulls (BW = 358 kg) were used. The treatments were as follows: mineral supplement (MS) alone and MS plus protein-energy supplement provided 3×, 5× and 7×/week. Supplementation frequency did not affect (P > 0.05) intake and digestibility. Average daily gain was greater (P < 0.001) to supplementation compared with MS. The supplementation 5×/week resulted in greater weight gain per hectare (9.24) and higher economic returns during the study period (1.64%) compared to other supplementations. Supplementation 5×/week increased animal performance and positively influenced economic returns.     

Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

Establishment of a comprehensive indicator system for the assessment of biodiversity and ecosystem services

Context

Common indicators are needed to monitor biodiversity loss and the implications for the sustainable provision of ecosystem services (ES). A plethora of different sets of indicators may hinder the identification of major endpoints for large-scale assessments of biodiversity and ecosystem services (BES).

Objectives

We aim to describe the main challenges of indicators for BES assessment and provide suggestions for establishing a comprehensive indicator system.

Methods

An extensive literature review was conducted in this study. We review the main challenges of indicators for BES assessment and propose corresponding improvements from our perspectives of theory and practical applications.

Results

The main theoretical challenges of existing indicators include inconsistent definitions and classifications of ES, misunderstanding of linkages among biodiversity, ecosystem structure, functions and services, and practical problems relate to such issues as indicator representativeness, data availability, and uncertainty. Our suggested improvements include clarifying the main terms, concepts and classification of indicators, establishing a robust conceptual framework with clear interactions among different components and indicators, selecting indicators based on ecosystem properties, streaming existing data into one platform, and strengthening validation of proxies. The steps for constructing a comprehensive indicator system for BES assessment are summarized.

Conclusions

Clear definitions of key terms are indispensable to classify indicators and construct a conceptual framework. Improved understanding of the relations among indicators of biodiversity, ecosystem functions, and ES across multiple scales can guide the development of the indicator system. The integrated indicator system is an important tool for BES assessment to support decision making for sustainable development.

     

Evaluation of the temporal distribution of <Emphasis Type="Italic">Fusarium graminearum</Emphasis> airborne inoculum above the wheat canopy and its relationship with Fusarium head blight and DON concentration

With the aim of unravelling the role of airborne Fusarium graminearum inoculum in the epidemic of Fusarium head blight (FHB) caused by this species in wheat spikes, a network of Burkard air samplers was set up in five wheat fields distributed in Belgium from 2011 to 2013. Each year from April to July, the daily amounts of F. graminearum inoculum above the wheat canopy were quantified using a newly developed TaqMan qPCR assay. The pattern of spore trapping observed was drastically different per year and per location with a frequency of detection between 9 and 66% and a mean daily concentration between 0.8 and 10.2 conidia-equivalent/m³. In one location, air was sampled for a whole year. Inoculum was frequently detected from the wheat stem elongation stage until the end of the harvesting period, but high inoculum levels were also observed during the fall. Using a window-pane analysis, different periods of time around wheat flowering (varying in length and starting date) were investigated for their importance in the relation between airborne inoculum and FHB parameters (FHB severity, frequency of F. graminearum infection and DON). For almost all the combinations of variables, strong and significant correlations were found for multiple window lengths and starting times. Inoculum quantities trapped around flowering were highly correlated with F. graminearum infection (up to R?=?0.84) and DON (up to R?=?0.9). Frequencies of detection were also well correlated with both of these parameters. DON concentrations at harvest could even be significantly associated with the F. graminearum inoculum trapped during periods finishing before the beginning of the anthesis (R?=?0.77). Overall, these results highlight the key role of the airborne inoculum in F. graminearum epidemics and underline the importance of monitoring it for the development of disease forecasting tools.     

Evaluation of the durability of the <Emphasis Type="Italic">Barley yellow dwarf virus</Emphasis>-resistant <Emphasis Type="Italic">Zhong ZH</Emphasis> and <Emphasis Type="Italic">TC14</Emphasis> wheat lines

Serial passage experiments (SPE) of a Barley yellow dwarf virus-PAV (BYDV-PAV) isolate were performed on Zhong ZH and TC14 wheat lines to evaluate the durability of their resistance to BYDV. At different passage numbers (from the 2nd to the 114th), biological properties of the produced isolates were recorded either by monitoring infection percentages and virus titers of the first 3 weeks of viral infection or by measuring their impact on yield components. Statistical analyses using the area under pathogen progress curves and the area under concentration progress curves demonstrated that these two resistant lines induce, after only a few passages, a selection of variant(s) with significantly modified infection abilities. Isolates resulting from SPE performed on these lines induced important decreases of yield components. These results indicate that the use of Zhong ZH and TC14 lines in BYDV-resistant breeding programmes should be approached with caution.     

Variability in response to toosendanin in a laboratory colony of<Emphasis Type="Italic">Trichoplusia ni</Emphasis>

Laboratory rearing of insects for scientific research ensures a constant supply of insects. However, laboratory colonies may diverge genetically and/or phenotypically from wild populations due to selection pressures in their artificial environment. This study examined how variation in a colony ofTrichoplusia ni influenced reproducibility over time of bioassays of toosendanin (a limonoid extracted from the bark ofMelia azedarach). In four of five experiments insect growth was reduced significantly (P=0.05) by the presence of toosendanin in the diet. The fifth experiment showed the same trend, but the effect was not significant. Mean larval mass of both control and treated insects also varied greatly among experiments. The considerable variability observed in this population highlights the importance of replicating experiments in time, and not just space, to confirm reproducibility of treatment effects. http://www.phytoparasitica.org posting Jan. 23, 2007.