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.     

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.     

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.     

Multi-scale study of bird species distribution and of their response to vegetation change: a Mediterranean example

Land use changes operate at different scales. They trigger a cascade of effects that simultaneously modify the composition or structure of the landscape and of the local vegetation. Mobil animals, and birds in particular, can respond quickly to such multi-scalar changes. We took advantage of a long term study on the response of songbirds to land-use changes on four Mediterranean islands in Corsica and Sardinia to explore the benefits of a multi-scale analysis of the relationships between songbird distribution, vegetation structure and landscape dynamics. Field data and aerial photographs were used to describe the vegetation at three different scales. Birds were censused by point counts. We used statistical variance decomposition to study how bird distribution and vegetation at various scales were linked. We analysed multi-scale vegetation changes (floristic composition, plot vegetation type, and landscape structure) and their consequences on bird distribution with multivariate and non-parametrical tests. The distribution of most species was linked to at least two spatial scales. The weight of a given scale was consistent with life-history traits for species whose biology was well-known. In the examples studied, vegetation composition, vegetation type and landscape changes that resulted from land abandonment negatively affected birds depending on open or heterogeneous areas. Our results emphasize that multi-scale analyses can greatly enhance our understanding of bird distribution and of their changes. Management of these populations should take into account measures at various spatial scales depending on the sensitivity of the species.     

Avian predation on wild and cultured sea urchin <Emphasis Type="Italic">Strongylocentrotus intermedius</Emphasis> in a rocky shore habitat

ABSTRACT:   The present study reports the annual variation in consumption of the sea urchin Strongylocentrotus intermedius by avian predators on a rocky shore where the culture of sea urchins has been conducted. Carrion crow and a few gull species were the most abundant avian predators and consumed a large number of sea urchins. Crows consumed mostly natural sea urchins, approximately 36 kg ww/ha per year on the intertidal rocky bench, but the gull species consumed mostly cultured sea urchins, approximately 100 kg ww/ha per year in the culture area. The seasonal variation in the amount of sea urchins consumed by crows was higher than that by the gull species, presumably because of the difference in foraging behavior in association with the seasonal tidal cycle. The natural sea urchins consumed are an allochthonous input from the subtidal to the intertidal habitat, and thus, crow predation may not affect the natural and the cultured populations of the sea urchin. The gull species consumed much of the cultured sea urchin, and thus, may be regarded as an effective predator causing damage to sea urchin culture. The results suggest that further studies are needed to determine why the gull species selectively feed on cultured sea urchins.