Cross-amplification of EST-derived markers among 16 grass species

The availability of a large number of expressed sequence tags (ESTs) has facilitated the development of molecular markers in members of the grass family. As these markers are derived from coding sequences, cross-species amplification and transferability is higher than for markers designed from genomic DNA sequences. In this study, 919 EST-based primers developed from seven grass species were assessed for their amplification across a diverse panel of 16 grass species including cereal, turf and forage crops. Out of the 919 primers tested, 89 successfully amplified DNA from one or more species and 340 primers generated PCR amplicons from at least half of the species in the panel. Only 5.2% of the primers tested produced clear amplicons in all 16 species. The majority of the primers (66.9%) were developed from tall fescue and rice and these two species showed amplification rate of 41.6% and 19.0% across the panel, respectively. The highest amplification rate was found for conserved-intron scanning primers (CISP) developed from pearl millet (91%) and sorghum (75%) EST sequences that aligned to rice sequences. The primers with successful amplification identified in this study showed promise in other grass species as demonstrated in differentiating a set of 13 clones of reed canary grass, a species for which very little genomic research has been done. Sequences from the amplified PCR fragments indicated the potential for the transferable CISP markers for comparative mapping purposes. These primer sets can be immediately used for within and across species mapping and will be especially useful for minor grass species with few or no available molecular markers.     

Idiopathic typhlocolitis in 40 aged donkeys

Typhlocolitis was diagnosed in 40 aged donkeys at routine post mortem examinations subjected to euthanasia for colic‐related clinical signs at The Donkey Sanctuary. Gross pathological changes included oedema, ulceration and haemorrhage involving the caecum and ventral colon. Histopathology indicated endoparasite and bacterial associated inflammation in 20 and 11 cases, respectively. Bacterial culture in 18 cases did not yield a definite aetiological agent. Other management and stress related factors were looked at to identify obvious risk factors. This report describes the clinical, biochemical and haematological parameters and pathological changes observed in 40 donkeys diagnosed with typhlocolitis.     

Micrometer and nanometer scale photopatterning of proteins on glass surfaces by photo-degradation of films formed from oligo(ethylene glycol) terminated silanes

Exposure of films formed by the adsorption of oligo(ethylene glycol) (OEG) functionalized trichlorosilanes on glass to UV light from a frequency-doubled argon ion laser (244 nm) causes photodegradation of the OEG chain. Although the rate of degradation is substantially slower than for monolayers of OEG terminated thiolates on gold, it is nevertheless possible to form micrometer-scale patterns by elective adsorption of streptavidin to exposed regions. A low density of aldehyde functional groups is produced, and this enables derivatization with nitrilotriacetic acid via an amine linker. Complexation with nickel enables the site-specific immobilization of histidine-tagged yellow and green fluorescent proteins. Nanometer-scale patterns may be fabricated using a Lloyd's mirror interferometer, with a sample and mirror set at right angles to each other. At low exposures, partial degradation of the OEG chains does not remove the protein-resistance of the surface, even though friction force microscopy reveals the formation of patterns. At an exposure of ca. 18 J cm(-2), the modified regions became adhesive to proteins in a narrow region ca. 30 nm (λ/8) wide. As the exposure is increased further the lines quickly broaden to ca. 90 nm. Adjustment of the angle between the sample and mirror enables the fabrication of lines of His-tagged green fluorescent protein at a period of 340 nm that could be resolved using a confocal microscope.     

Quantifying rice yield gaps and their causes in Eastern and Southern Africa

The demand for rice in Eastern and Southern Africa is rapidly increasing because of changes in consumer preferences and urbanization. However, local rice production lags behind consumption, mainly due to low yield levels. In order to set priorities for research and development aimed at improving rice productivity, there is a need to characterize the rice production environments, to quantify rice yield gaps—that is, the difference between average on-farm yield and the best farmers’ yield—and to identify causes of yield gaps. Such information will help identifying and targeting technologies to alleviate the main constraints, and consequently to reduce existing yield gaps. Yield gap surveys were conducted on 357 rice farms at eight sites (19–50 farmers per site) across five rice-producing countries in Eastern and Southern Africa—that is Ethiopia, Madagascar, Rwanda, Tanzania and Uganda—for one or two years (2012–13) to collect both quantitative and qualitative data at field and farm level. Average farm yields measured at the eight sites ranged from 1.8 to 4.3 t/ha and the average yield gap ranged from 0.8 to 3.4 t/ha. Across rice-growing environments, major causes for yield variability were straw management, weeding frequency, growth duration of the variety, weed cover, fertilizer (mineral and organic) application frequency, levelling and iron toxicity. Land levelling increased the yield by 0.74 t/ha, bird control increased the yield by 1.44 t/ha, and sub-optimal management of weeds reduced the yield by 3.6 to 4.4 t/ha. There is great potential to reduce the current rice yield gap in ESA, by focusing on improvements of those crop management practices that address the main site-specific causes for sub-optimal yields.     

Prevalence, bacterial causes, and antimicrobial susceptibility profile of mastitis isolates from cows in large-scale dairy farms of Northern Ethiopia

The study was undertaken to determine the prevalence of bovine mastitis, isolate mastitis causing bacteria, assess the association of some risk factors, and determine the antibiotic resistance pattern of bacterial isolates in cows in large-scale dairy farms of Northern Ethiopia. A total of 305 lactating and nonlactating cows were included in the present study. The overall prevalence of clinical and subclinical mastitis was 3.6 and 33.8?%, respectively. The quarter level prevalence was 15.4?%; from which, 11.9 and 1.1?% were subclinical form and blind teat, respectively, while the remaining 2.4?% were of clinical form. Staphylococcus aureus accounted for 36?% of the isolates followed by Escherichia coli (27.3?%). Risk factors including age (p?9?years (OR?=?3.63, 95?% CI?=?1.42-9.25)), cows with four to seven calves (OR?=?3.39, 95?% CI?=?2.06-5.60), and cows in late lactation stage (OR?=?3.79, 95?% CI?=?1.64-8.75). In multivariable logistic regression analysis, age (p?=?0.005) and lactation stage (p?=?0.027) showed statistically significant association with the occurrence of mastitis. The antimicrobial susceptibility pattern showed high susceptibility of S. aureus to nalidixic acid (82.4?%) followed by chloramphenicol (58.8?%); however, these species were resistant to the rest of the antimicrobials tested. Highest resistance was observed against clindamycin and ampicillin. Coliform bacteria (E. coli and Klebsiella pneumoniae) showed resistance to most of the antimicrobials used. Detailed investigation is needed to identify the interplay of managemental and environmental risk factors to design appropriate control measures.     

Prevalence and species composition of ixodid ticks infesting horses in three agroecologies in central Oromia, Ethiopia

A cross-sectional study was conducted to determine the species composition and prevalence of ixodid ticks infesting horses in three agroecological zones in central Ethiopia. For this purpose, a total of 1,168 horses were examined for tick infestation. An overall prevalence of 39.04% of tick infestation on horses was recorded. A total of 917 adult ticks were collected from infested horses. Amblyomma, Boophilus, Rhipicephalus, and Hyalomma genera with the respective prevalence of 3.2%, 1.8%, 29.2%, and 4.7% were identified. In the study, Rhipicephalus evertsi evertsi was encountered with the highest prevalence (15.8%) whereas Amblyomma gemma was with lowest prevalence (1.5%). From the highland, Hyalomma marginatum rufipes (3.1%), Hyalomma truncatum (1.0%), and Boophilus decoloratus (0.3%) were identified. From the midland, R. evertsi evertsi (27.5%), Rhipicephalus pulchellus (18%), Amblyomma variegatum (3.6%), B. decoloratus (2.8%), H. marginatum rufipes (2.6%), H. truncatum (1.8%), and A. gemma (1.5%) were identified. R. evertsi evertsi, 107 (27.5%), was with the highest prevalence in the midland. From the lowland, R. pulchellus (22.3%), R. evertsi evertsi (20%), H. truncatum (3.6%), A. gemma (3.1%), B. decoloratus (2.3%), H. marginatum rufipes (2.1%), and A. variegatum (1.5%) were identified. In the lowland, R. pulchellus, 87 (22.3%), was the most abundant tick species. The overall prevalence of tick infestation on horses was significantly (P < 0.05) higher both in the midland, 225 (57.8%), and the lowland, 214 (54.87%), than the highland, 17 (4.4%). This suggests that horses in midland and lowland are at higher risk of tick infestation than those horses in the highland. Further studies on the role of ticks in transmission of diseases to equines and the importance of horses as alternative hosts in different parts of Ethiopia are needed.     

Zymogram patterns of α-amylase isozymes inEthiopian tetraploid wheat landraces: insight into their evolutionaryhistory and evidence for gene flow

The variation for -amylaseisozymes was analyzed in 71 tetraploid wheat (AABB genome)landraces from Ethiopia, including accessions of Triticumdicoccon Shrank, T.turgidum L., T.durum Desf., T.pyramidale Percival and T.aethiopicum Jakubz., by thin-layerpolyacrylamide gel isoelectric focusing. Four zymogram phenotypeswere obtained from all the materials studied. Except inT. dicoccon, however, thestandard pattern of the durum wheat variety, LD222, was predominant.T. dicoccon showed twozymogram types that differed for the absence or consistently weakactivity of band 18(-Amy-B1)of the malt type. Band 1(-Amy-B3)of the malt type was fixed in T.dicoccon and present in only 10% of thefree-threshing (FT) types. Generally, the resultsindicated that the variation for-amylase isozymes in cultivatedtetraploid wheats, including landraces from secondary centers, is lowpossibly due to the founder effect or as a result of selection. Thedata were useful for inferences about the evolutionary history ofEthiopian wheat landraces. It is speculated thatT. dicoccon was the firstwheat to arrive in the Ethiopian highlands ca.5000 years ago. However, it is not known whether the present day FTEthiopian tetraploid wheat landraces are direct descendants ofT. dicoccon, or whetherthey were introduced independently. The clear differences in-amylase zymogram patterns favor the latterhypothesis. However, the presence of band1 in some of the FT types indicated the occurrence of geneflow between the FT types and T.dicoccon, which may also explain thequantitative nature of spike threshability in these landraces.Possible implications for durum wheat breeding arediscussed.     

A survey of seasonal patterns in strongyle faecal worm egg counts of working equids of the central midlands and lowlands, Ethiopia

A study was conducted for two consecutive years (1998–1999) to determine the seasonal patterns of strongyle infection in working donkeys of Ethiopia. For the purpose 2385 donkeys from midland and lowland areas were examined for the presence of parasitic ova. A hundred percent prevalence of strongyle infection with similar seasonal pattern of strongyle faecal worm egg output was obtained in all study areas. However, seasonal variations in the number of strongyle faecal worm egg output were observed in all areas. The highest mean faecal worm egg outputs were recorded during the main rainy season (June to October) in both years in all areas. Although an increase in the mean strongyle faecal egg output was obtained in the short rainy season (March–April) followed by a drop in the short dry season (May), there was no statistically significant difference between the short rainy season and long dry season (Nov–Feb) (P > 0.05). A statistically significant difference however, was obtained between the main rainy season and short rainy season, and between the main rainy season and dry season (P < 0.05). Based on the results obtained it is suggested that the most economical and effective control of strongyles can be achieved by strategic deworming programme during the hot dry pre-main rainy season (May), when the herbage coverage is scarce and helminthologically ‘sterile’, and the arrested development of the parasites is suppose to be terminating. This could insure the greatest proportion of the existing worm population to be exposed to anthelmintic and also reduces pasture contamination and further infection in the subsequent wet season.     

Cropping sequence and nitrogen fertilizer effects on the productivity and quality of malting barley and soil fertility in the Ethiopian highlands

The productivity and quality of malting barley were evaluated using factorial combinations of four preceding crops (faba bean, field pea, rapeseed, and barley) as main plots and four nitrogen fertilizer rates (0, 18, 36, and 54 kg N ha^?1) as sub-plots with three replications at two sites on Nitisols of the Ethiopian highlands in 2010 and 2011 cropping seasons. Preceding crops other than barley and N fertilizer significantly improved yield and quality of malting barley. The highest grain yield, kernel plumpness, protein content, and sieve test were obtained for malting barley grown after faba bean, followed by rapeseed and field pea. Nitrogen fertilizer significantly increased yield, protein content, and sieve test of malting barley. All protein contents were within the acceptable range for malting quality. Inclusion of legumes in the rotation also improved soil fertility through increases in soil carbon and nitrogen content. We conclude that to maximize yield and quality of malting barley, it is critical to consider the preceding crop and soil nitrogen status. Use of appropriate break crops may substitute or reduce the amount of mineral N fertilizer required for the production of malting barley at least for one season without affecting its quality.     

In vitro pre-exposure of Haemonchus contortus L3 to blood eosinophils reduces their establishment potential in sheep

Different authors have reported that eosinophils are capable of immobilising infective larvae of different species of nematodes in vitro. However, classifying larvae as mobile or immobile is so subjective that it does not always mean all apparently immobile larvae are dead or those that are mobile are capable of surviving further immune responses if administered to their natural hosts. The objective of this experimental study was therefore to substantiate the role of eosinophils in the killing of Haemonchus contortus infective larvae by comparing the infectivity in sheep of larvae that had been incubated with eosinophil-enriched cell suspensions with control larvae. Since it was not possible to isolate pure eosinophils from sheep blood, we were compelled to evaluate the effects of other blood cells contaminating our eosinophil-enriched suspensions. Although eosinophils and neutrophils were the only cells found adherent to H. contortus infective larvae in vitro, induced eosinophils in the presence of immune serum were primarily responsible for the drastic reduction in larval motility compared to the minor effects of neutrophils and mononuclear cells. Corresponding reductions in faecal egg count and worm numbers were observed when the incubated larvae were transferred intra-abomasally to sheep. Interestingly, the proportion of larvae that failed to establish was much higher following incubation with induced eosinophils compared with other cells or with immune serum alone. Although this study did not address the in vivo role of eosinophils in sheep, the results strongly indicate that sheep blood eosinophils have a larval killing potential in vitro, and a larval mobility test alone may not fully explain the level of damage inflicted on the larvae.