PRRSV2 genetic diversity defined by RFLP patterns in the United States from 2007 to 2019

The genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) increases over time. In 1998, restriction-fragment length polymorphism (RFLP) pattern analysis was introduced to differentiate PRRSV wild-type strains from VR2332, a reference strain from which a commercial vaccine (Ingelvac PRRS MLV) was derived. We have characterized here the PRRSV genetic diversity within selected RFLP families over time and U.S. geographic space, using available ISU-VDL data from 2007 to 2019. The 40,454 ORF5 sequences recovered corresponded to 228 distinct RFLPs. Four RFLPs [2-5-2 (21.2%), 1-7-4 (15.6%), 1-4-4 (11.8%), and 1-8-4 (9.9%)] represented 58.5% of all ORF5 sequences and were used for cluster analysis. Over time, there was increased detection of RFLPs 2-5-2, 1-7-4, 1-3-4, 1-3-2, and 1-12-4; decreased detection of 1-4-2, 1-18-4, 1-18-2, and 1-2-2; and different detection trends for 1-8-4, 1-4-4, 1-26-1, 1-22-2, and 1-2-4. An over-time cluster analysis revealed a single cluster for RFLP 2-5-2, supporting that sequences within RFLP 2-5-2 are still relatively conserved. For 1-7-4, 1-4-4, and 1-8-4, there were multiple clusters. State-wise cluster analysis demonstrated 4 main clusters for RFLP 1-7-4 and 1-8-4, and 6 for RFLP 1-4-4. For the other RFLPs, there was a significant genetic difference within them, particularly between states. RFLP typing is limited in its ability to discriminate among different strains of PRRSV. Understanding the magnitude of genetic divergence within RFLPs helps develop PRRSV regional control programs, placement, herd immunization strategies, and design of appropriate animal movements across borders to minimize the risk of PRRSV transmission.     

Identification and isolation of a new x-type HMW glutenin subunit 1Dx1.6t gene from Aegilops tauschii

A new x-type HMW glutenin subunit, designated as 1Dx1.6^t from Aegilops tauschii was identified and characterized by SDS-PAGE and MALDI-TOF-MS. This subunit is located between 1Dx2 and 1Dx1.5^t and possesses a molecular mass ( M _r) of 88565.8 Da. Its complete coding sequence was amplified via allele-specific PCR (AS-PCR), and the amplified product was cloned and sequenced. The authenticity of the cloned 1Dx1.6 ^t gene was confirmed by successful expression of its open reading frame in Escherichia coli. The molecular characterization of 1Dx1.6 ^t gene showed that its coding region consisted of 2541 bp encoding a polypeptide of 845 amino acid residues. Sequence comparison to previously characterized 1Dx1.5^t subunit which is related to good dough quality of bread wheat indicated that the 1Dx1.6^t subunit displayed high homology, but possesses 14 residue substitutions and a nonapeptide insertion. A total of 12 single-nucleotide polymorphisms (1 per 212 bp) was identified in the 1Dx1.6 ^t allele (11 in repetitive domain and 1 in the C-terminal domain), which could facilitate the design of AS-PCR markers.     

Sorte,Boden und die Mineralstoffe im Weizen

Zusammenfassung Seit mehreren Jahren sind an der Wiener Landw.-chemischen Bundesversuchsanstalt in Wien Untersuchungen im Gange, die zeigen sollen, ob und wie der Mineralstoffgehalt von Weizen sortenabhängig ist und wie der Mineralstoffgehalt verschiedener Sorten vom Düngungszustand und sonstigen bodenchemischen Faktoren abhängt. Auch wurde geprüft, wie sich der Mineralstoffgehalt in verschiedenen Teilen der Pflanze und in der ganzen Pflanze im Laufe der Vegetationsperiode verändert. Hierbei ist es notwendig, die Ergebnisse auf eine während der ganzen Vegetationszeit gleichbleibende Bezugsgröße zu beziehen. Trockengewicht oder Aschegehalt kommen daher nicht in Frage. Als geeignete Bezugsgrößen erwiesen sich die durchschnittliche Einzelpflanze, der Gesamtentzug pro Flächeneinheit, aber auch der Mineralstoffgehalt eines Saatkornes scheint brauchbar zu sein.Die Untersuchungen sind noch keineswegs abgeschlossen, ein Teil des vorhandenen außerordentlich umfangreichen Zahlenmaterials, das sich sowohl auf die Massenelemente als zum Teil auch auf die Spurenelemente bezieht, konnte aber schon ausgewertet werden und wird in übersichtlicher Form dargestellt. Der verhältnismäßig geringe Informationsgehalt der auf Trockensubstanz bezogenen Zahlen kommt hierbei deutlich zum Ausdruck und Luxuskonsum und sortenabhängiges Speicher- und Aufnahmevermögen werden deutlich.

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Summary The Federal Institute for Agricultural Chemistry in Vienna has for a number of years investigated the mineral composition of different varieties of wheat and the influence of the nutritional status and other chemical factors of the soil on the minerals in the wheat plant. In addition, the mineral composition of the wheat plants has been investigated as to how it is dependent on the developmental state and growth of the plants. To this end it was necessary to use a point of reference which is constant during all of the growing period. Dry matter and ash are therefore not suitable for this purpose. Useful points of reference were found to be the average plant, the total nutrient uptake per unit area or the mineral content of the seed used.The investigations are by no means yet completed but some of the many data so far obtained, have been analysed. They concern not only the major but also a number of minor elements and they are presented in a summarized form. It can be seen that comparatively little information can be obtained from data relating the mineral content of the plants to their dry matter content and that luxury consumption and varietal differences of nutrient uptake and nutrient storage come to light, using the suggested points of reference.

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Résumé Depuis plusieurs années à la station fédérale de chimie agricole de Vienne des recherches sont en cours pour déterminer dans quelle mesure la teneur en substances minérales du blé depend des variétés, de la fumure et des facteurs agropédologiques. Les variations taux de substances minérales dans les diverses parties de la plante, pendant de la phase de la développement de la végétation, ont également été suivies. Il est nécessaire pour cela de rapporter les résultats à une valeur de référence qui reste constante durant toute la période de végétation. Le poids sec le taux de cendres, ne peuvent servir de référence. Il s'est révélé objectif de rapporter les résultats à la plantule moyenne, et de ramener les prélèvements de minéraux à la surface cultivée; le taux de substances minérales d'un grain de semence est aussi une référence utilisable.Nos recherches ne sont nullement terminées; une partie de nos très nombreux résultats numériques, qui se rapportent aussi bien aux macroéléments qu'à divers oligoéléments, ont été exploités et présentés de facon synthétique. L'information pauvre de la valeur relative des taux rapportés au poids sec apparaît ainsi; on peut mettre en évidence une consommation de luxe, ainsi qu'une différence dans le pouvoir d'absorption et de stockage selon la variété.

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Vortrag gehalten auf der gemeinsamen Tagung EUCARPIA-CIQ in Lund, Schweden (14. – 17. Juli 1965) am 16.7.1965.     

Phenotypic Characterization of Canine Intestinal Intraepithelial Lymphocytes in Dogs with Inflammatory Bowel Disease

Background

Many dogs suffering from inflammatory bowel disease (IBD) are presented to veterinary clinics. These patients are diagnosed based on a history of chronic gastrointestinal signs and biopsy‐confirmed histopathologic intestinal inflammation. Intestinal intraepithelial lymphocytes (IEL) are part of the first line of defense in the gastrointestinal immune system. Alterations in IEL subsets may play a role in the pathogenesis of IBD.

Hypothesis

The aim of this study was to characterize the phenotypes of IEL in dogs with IBD compared with healthy control dogs.

Animals

Intestinal intraepithelial lymphocytes subpopulations of control dogs (n = 5) obtained from endoscopic biopsies (EB) were compared to those obtained from full thickness biopsies (FTB) on the same day. In addition, the phenotypes of IEL from FTB of control dogs (n = 10) were compared with EB of IBD dogs (n = 10). Each participant was scored clinically using the canine inflammatory bowel disease activity index (CIBDAI), and all samples were graded histopathologically. Three‐color flow cytometry of isolated IEL was performed using monoclonal antibodies against T‐ and B‐lymphocyte subpopulations.

Results

No significant differences in the composition of IEL subpopulations were found in control dogs based on method of biopsy. The IBD dogs had significantly higher CIBDAI and histopathologic scores compared with control dogs and their IEL contained a significantly higher frequency TCRγδ T‐cells.

Conclusions and Clinical Importance

Endoscopic biopsies provide suitable samples for 3‐color flow cytometry when studying canine intestinal IEL and IBD patients show significant changes of major T‐cell subsets compared to healthy control dogs.     

Influence of elevated protein and tannin-rich peanut skin supplementation on growth performance,blood metabolites,carcass traits and immune-related gene expression of grazing meat goats

The aim of the present study was to define whether elevated rumen-undegradable protein (RUP) and tannin-rich peanut skin (PS) supplementation would affect animal growth performance, average daily gain (ADG), blood metabolites, carcass traits associated with lipogenic and immune-related gene expressions in meat goats grazing winter wheat (WW). Thirty-six Kiko-crossbreed male goats at approximately 6 months of age were blocked by body weight (BW; 25.6 ± 1.1 kg) and randomly assigned to one of the four treatments with two replicates based on a 2 × 2 factorial design. Diets contained PS replacing alfalfa meal (ALM), without or with RUP supplementation. Both PS and ALM were incorporated into grain mix portion of the diet and pelletized, with remaining diets fed ad libitum of WW forage for a period of 51 days. Lipogenic genes examined included SCD, ACLY, YWHAZ, PPIA and FABP4, while immune-related genes examined included ACTB (as a control gene), H3F3A, PPIA, IRF3, STAT2, HERC3 and IFIT3 antibody genes. The meat goats on PS-pellet-supplemented group with or without RUP supplementation grew 38.5% faster ADG (p < .001) when compared to control-supplemented group. When goats received PS diet, empty body weight, hot carcass, cold carcass, shoulder, hind shank, rack, loin and fat thickness were greater (p < .05) than control diet. Animals on PS-pellet had higher ACLY, YWHAZ, PPIA and FABP4 gene expression (p < .05) when compared to ALM-pellet control, with RUP by PS-pellet interactions (p < .01). Goats receiving additional RUP supplementation had increased (p < .05) STAT2 gene expression, whereas goats receiving PS-pellet supplementation showed increased STAT2 (p < .05) and a tendency to increase IRF3 (p = .07) gene expressions. In conclusion, the addition of PS-pellet or RUP supplementation has the potential to improve ADG and altered selected lipogenic and immune-related gene expressions.     

Characterization of a PCR‐based lymphocyte clonality assay as a complementary tool for the diagnosis of feline lymphoma

Differentiation between resident mature lymphocyte populations and small cell lymphoma cannot be made by cytological review alone and remains challenging in histopathological review. These cases warrant application of complementary tools like PCR‐based immunoglobulin (IG) and T‐cell receptor (TCR) clonality testing for confirmation. In this prospective study, diagnostic sensitivity and specificity of different primer sets for routine diagnosis of feline TCR gamma (TCRG) and complete IG heavy chain (IGH) gene rearrangements were assessed. Fine needle aspirates from 20 feline lymphoma cases and lymph node material from 10 cats without hematopoietic neoplasia were subjected to clonality testing. Feline lymphoma cell lines and previously confirmed patient material served as positive control. Detection limits for clonal populations within a polyclonal background was 90% for B‐cells and 50% for T‐cells. Diagnostic sensitivity and specificity of the clonality assay were 70% and 90%. Overall diagnostic accuracy was 77%, positive predictive value 93% and negative predictive value 60%.     

Amplified Fragment Length Polymorphism Diversity in Cephalosporium maydis from Egypt

ABSTRACT Cephalosporium maydis, the causal agent of late wilt of maize, was first described in Egypt in the 1960s, where it can cause yield losses of up to 40% in susceptible plantings. We characterized 866 isolates of C. maydis collected from 14 governates in Egypt, 7 in the Nile River Delta and 7 in southern (Middle and Upper) Egypt, with amplified fragment length polymorphism (AFLP) markers. The four AFLP primer-pair combinations generated 68 bands, 25 of which were polymorphic, resulting in 52 clonal haplotypes that clustered the 866 isolates into four phylogenetic lineages. Three lineages were found in both the Nile River Delta and southern Egypt. Lineage IV, the most diverse group (20 haplotypes), was recovered only from governates in the Nile River Delta. In some locations, one lineage dominated (up to 98% of the isolates recovered) and, from some fields, only a single haplotype was recovered. Under field conditions in Egypt, there is no evidence that C. maydis reproduces sexually. The nonuniform geographic distribution of the pathogen lineages within the country could be due to differences in climate or in the farming system, because host material differs in susceptibility and C. maydis lineages differ in pathogenicity.