Seroprevalence of bovine leukemia virus in dairy cattle in Argentina: comparison of sensitivity and specificity of different detection methods

Bovine leukemia virus (BLV) is a retrovirus that induces a chronic infection in cattle, which develop in three possible pathological forms: asymptomatic course, persistent lymphocytosis (PL) and lymphosarcoma. Once infected, cattle remain virus carriers for life and start to show a serological reaction within a few weeks after infection. Eradication and control of the disease is based on early diagnostic and segregation of the carriers. The agar gel immunodiffusion (AGID) test has been the serological test of choice for routine diagnosis of serum samples. Nevertheless, in more recent years, the enzyme-linked immunosorbent assay (ELISA) has replaced the AGID for large scale testing. Although Argentina has over 60 million cattle population, no nationwide studies have been conducted yet to determine the prevalence of the infection. To estimate the rate of BLV infection in dairy cattle in Argentina, a survey for specific antibodies in >10,000 serum samples from animals over 18 months old, belonging to 363 different herds from the largest dairy production areas of the country, was carried out in our laboratory, along 1999. For this purpose, we developed an ELISA to detect serum antibodies against the BLV virus. The cut-off of the ELISA was established over 339 serum samples, using polymerase chain reaction and southern blot (PCR-SB) as confirmatory test. The sensitivity and specificity of the ELISA was of 97.2 and 97.5%, respectively, while the local official AGID test showed a sensitivity of 79.7% and specificity of 99.0%. To know the seroprevalence of BLV on dairy herds, and also the incidence of the infection within the herd, the serological survey was based on individual serum samples. The results show that the prevalence of infected individuals is of 32.85%, while the percentage of infected herds, harboring one or more infected animals, is of 84%. These results indicate a medium level of seropositive animals when taken individually, but a high prevalence of infected farms, which has been notoriously increased in the last 15 years as shown when compared with previous data from particular geographic areas, indicating that BLV constitutes a serious sanitary problem for dairy producers in Argentina. They also indicate the poor sensitivity of the official AGID test used in the country.     

Effects of diet and feeding time on daily variations in plasma insulin,hepatic c-AMP and other metabolites in a teleost fish,Dicentrarchus labrax L.

Juvenile sea bass, 1.5 years old, of mixed sex, held on long photoperiods were fed early on the photoperiod and late on the photoperiod, using different diets. Fish fed natural diets showed a daily rhythmicity of plasma insulin, liver c-AMP, plasma glucose, liver glycogen and muscle glycogen content, however, fish fed a commercial diet did not show this daily rhythmicity except for plasma insulin levels. In addition, these fish had significantly lower levels of plasma insulin, liver c-AMP and plasma glycerol than the group fed on the natural diets at similar feeding times. The time of feeding also induced different rhythmicity patterns in hormones and metabolites as well as a significant change in their mean levels. These facts are discussed in relation with the pre-feeding activity and increased appetite exhibited by the fish fed late on the photoperiod and with their implications on fish culture.     

Annual cycle of plasma insulin and glucose of sea bass.Dicentrarchus labrax,L.

Annual plasma insulin and glucose cycles were studied inDicentrarchus labrax maintained in either seawater (37.8) or brackish water (3.5). In both media, the highest insulin levels were found during the prespawning period (August–November) coincident to increases in weight and a decrease in plasma glucose. During spawning (December–April) and postspawning (May–July) periods, the decrease in insulin occurred at the same time as a reduction in growth and an increase of plasma glucose. Temperature and salinity conditions impeded spawning in the brackish water group, in which a minor weight loss was regained more quickly than in the sea water group; insulin levels were also higher.     

Linkage map of prolamin loci Gli-D4 and Gli-D5 in hexaploid wheat

By electrophoretic analysis of F2 progenies from crosses among the hexaploid wheat varieties ‘Cajeme 71’, ‘Ablaca’, ‘Anza’ and ‘Pané 247’, two new prolamin loci Gli-D4 and Gli-D5, were mapped on the short arm of chromosome ID. The Gli-D4 locus controls gliadins of type γ and is situated on the short arm of chromosome ID between the centromere and the Gli-D1 locus with a genetic distance of 10.1±2.4 cM from this locus. The Gli-D5 locus controls gliadin type ω and was mapped 3.7 ± 0.8 cM from Gli-Dl and located between Gli-Dl and the telomere.     

Biochemical and molecular characterisation of Glu-1 loci in Argentinean wheat cultivars

The high molecular weight glutenin subunit (HMW-GS) composition of acollection of 107 Argentinean bread wheat cultivars was analysed bysodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE).Allelic variation at the Glu-1 loci was identified and its frequencycalculated. Eleven alleles were detected, three encoded at the Glu-A1locus, six at the Glu-B1 locus and two at the Glu-D1 locus. Alow frequency of the null allele at the Glu-A1 locus and a highfrequency of subunits 5+10 at the Glu-D1 locus were observed.Reversed phase-high performance liquid chromatography (RP-HPLC)analysis was used to further characterise HMW-GS. Two different types ofBx subunit 8 (named subunits 8 and 8) were detected, the latterhaving shorter elution time. Subunit 8 was not identifiable bySDS-PAGE. According to quantification by RP-HPLC analysis, two groupsof subunit 7 were observed. One group, with a relatively high proportionof subunit 7 (approximately 39% of the total amount of HMW-GS)appeared in cultivars with allele 7+8 at the Glu-B1 locus; asecond group of subunit 7 (around 24% of the total amount ofHMW-GS), was found in alleles 7+8, 7+8 and 7+9. Restrictionfragment length polymorphisms (RFLP) analyses of HMW-GS genes werealso carried out after digestion of genomic DNA with HindIII andTaqI restriction enzymes. The relationship between DNA fragment sizeand glutenin subunits, as estimated by electrophoretic mobility inSDS-PAGE, was also examined. The restriction enzyme TaqIdemonstrated to be a useful tool to detect homozygous plants in selectionprograms against the Glu-A1 null allele.     

Waxy proteins and amylose content in diploid Triticeae species with genomes A, S and D

A collection of 89 accessions of diploid species of wheat was analysed for waxy protein in the grain: 39 accessions of Einkorn wheats, 41 accessions of Sitopsis section wheat and nine accessions of Triticum tauschii. The electrophoretic patterns showed low polymorphism. In each group of wheat, a single and different allele was detected. In accessions of Einkorn wheats that allele had a similar electrophoretical mobility to the Wx‐A1a allele of the bread wheat ‘Chinese Spring’, in accessions of the Sitopsis section it had a similar mobility to that of the Wx‐B1f allele of tetraploid wheat, and in the accessions of T. tauschii, it was similar to the Wx‐D1a allele of the bread wheat ‘Chinese Spring’. The accessions were also analysed for apparent amylose content. Results showed that amylose content ranged from 22 to 35% in Einkorn wheats, from 28 to 41% in the Sitopsis section and from 26 to 35% in accessions of T. tauschii.     

Polymorphism of high molecular weight glutenin subunits in three species of Aegilops

A collection of 136 accessions of Aegilops umbellulata (39), Ae. comosa (75) and Ae. markgrafii (22) was analysed for high-molecular-weight (HMW) glutenin subunits composition. The homogeneity of the accessions was studied and 55.1% of the collection was homogeneous for HMW glutenin subunits (29 Ae. umbellulata, 33 Ae. comosa and 14 Ae. markgrafii). The HMW glutenin subunits of Ae. umbellulata are encoded by the Glu-U1 locus; in Ae. comosa results showed that this proteins are encoded at the 1M chromosome, and the locus was named Glu-M1. In Ae. markgrafii it was assumed that HMW glutenin subunits were encoded by an homoeologous locus and it was named Glu-C1. All the accessions of Ae. umbellulata and Ae. markgrafii expressed both, x-type and y-type subunits. Among the Ae. comosa accessions, only one expressed an x-type subunit alone. All the accessions of Ae. umbellulata and some of Ae. comosa had x-type glutenins of higher molecular weights than those commonly present in bread wheat. A total of 8 alleles were detected at the Glu-U1 locus, 11 at the Glu-M1 and 4 at the Glu-C1. The new HMW glutenin variation found in this work suggests their possible utilisation in breeding for wheat quality.     

Cumulative and interaction effects of prolamin allelic variation and of 1BL/1RS translocation on flour quality in bread wheat

The allelic variation of prolamin loci was studied in three F₂ progenies from three crosses between the 1BL/1RS cultivar Triana and Yécora Rojo, Pavón and Florence Aurora, cultivars without the translocation. According to the 1:2:1 theoretical proportions observed in the allelic variants of the Glu-B3/Gli-B1 loci of the parent without the translocation, the inheritance as a block of the rye chromosome arm was confirmed. A group of F₃-F₄ recombinant lines, developed from these crosses was evaluated using the SDS-sedimentation test and the mixograph and alveograph tests. The presence of the 1BL/1RS translocation was not associated with significantly lower grain protein content values or with the optimum mixing time in the mixograph of the genotypes. The effect of the 1BL/1RS translocation on most of the quality parameters was highly dependent on the genetic pool. Significant increases in gluten strength and better mixing properties associated with the presence of some alleles of the Glu-A1, Glu-A3/ Gli-A1 and Gli-D2 loci were detected. The additivity and the interaction of prolamin gene effects with the rye translocation in the 1BL/1RS lines and its possible use in plant breeding are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.