Comparison of polyacrylamide gel electrophoresis, an enzyme-linked-immunosorbent assay, and an agglutination test for the direct identification of bovine rotavirus from feces and coelectrophoresis of viral RNAs

The dsRNA concentrated polyacrylamide gel electrophoresis (CPAGE) detected rotavirus directly from 19% of 77 stool specimens from diarrheic calves. A commercial enzyme-linked immunosorbent assay (ELISA) detected 25%, latex agglutination test, 23%, and polyacrylamide gel electrophoresis (PAGE), 19%. Establishing CPAGE as the "standard," the commercial ELISA and the latex agglutination test both had higher sensitivity (84%) than PAGE (79%). However, PAGE produced the highest specificity (100%), followed by agglutination (88%) and ELISA (84%). The commercial ELISA had a slightly higher sensitivity than agglutination, PAGE, and CPAGE, but the ELISA specificity was generally lower. The latex agglutination test had a lower sensitivity than ELISA, but specificity was higher. Agglutination had similar negative predictive values (94%), compared with agglutination and PAGe, but had the lowest positive predictive value (a measure of accuracy) (70%). Agreement with CPAGE was highest for PAGE (94.8%), followed by agglutination (87%) and ELISA (84.4%). The calculated percentages of total disagreement with all other tests indicated that ELISA differed from the other rotavirus detection assays in 10.4% of the cases, agglutination in 7.8%, PAGE in 2.6%, and CPAGE in 1.3%. The 2 PAGE assays allowed the detection of atypical rotaviruses from feces based on the characteristic "super-short" migration pattern of the 11 genomic segments of rotaviruses and of other members of the Reoviridae.     

Spatial distribution and habitat selection of culicoides imicola: The potential vector of bluetongue virus in Tunisia

The increasing threat of vector-borne diseases (VBDs) represents a great challenge to those who manage public and animal health. Determining the spatial distribution of arthropod vector species is an essential step in studying the risk of transmission of a vector-borne pathogen (VBP) and in estimating risk levels of VBD. Risk maps allow better targeting surveillance and help in designing control measures. We aimed to study the geographical distribution of Culicoides imicola, the main competent vector of Bluetongue virus (BTV) in sheep in Tunisia. Fifty-three records covering the whole distribution range of C.imicola in Tunisia were obtained during a 2-year field entomological survey (August 2017 – January 2018 and August 2018 – January 2019). The ecological niche of C. imicola is described using ecological-niche factor analysis (ENFA) and Mahalanobis distances factor analysis (MADIFA). An environmental suitability map (ESM) was developed by MaxEnt software to map the optimal habitat under the current climate background. The MaxEnt model was highly accurate with a statistically significant area under curve (AUC) value of 0.941. The location of the potential distribution of C. imicola is predicted in specified regions of Tunisia. Our findings can be applied in various ways such as surveillance and control program of BTV in Tunisia.     

Epistasis and Maternal Effect in Resistance to Puccinia coronata Cda. f. sp. avenae Eriks in Oats （Avena sp.）

The objective of this paper was to investigate the mode of heredity for resistance in oats （Avena sp.） to crown rust caused by Puccinia coronata Cda. f. sp. avenae Eriks, Eight generations of 2 crosses were used to estimate genetic effects and narrow-sense heritability （NSH）. Separate generation means analysis （GMA） indicated a complex gene action controlling this trait with additive, dominance, epistatic and maternal effects （ME）. The genetic model which assumed no epistasis and no ME did not accurately describe the resistance to P. coronata. In both crosses, the digenic epistatic model with ME was sufficient to explain variation in generation means for isolate CRec58 and isolate CRec46. Additive dominance and epistatic components were negative in most cases, suggesting that gene effects contributed more to the resistance than to the susceptibility. The estimated values of NSH were 15-99% depending upon the cross and isolates. The results indicated that appropriate choice of maternal parent and recurrent selection would increase resistance to crown rust in oats.     

Analysis of genetic variation of epizootic hemorrhagic disease virus and bluetongue virus field isolates by coelectrophoresis of their double-stranded RNA.

Thirty-two bovine field isolates of bluetongue virus (BTV), 6 field isolates of epizootic hemorrhagic disease virus (EHDV) from deer, 4 BTV prototype serotypes (10, 11, 13, and 17), and 2 EHDV prototype serotypes (1 and 2) were coelectrophoresed, using polyacrylamide gels. Field isolates were obtained from various regions of the United States. Analysis of polyacrylamide gels and scattered plots generated for comparison of migration patterns for different isolates within each serotype of BTV revealed wide variation among the individual segments. The BTV serotypes 10 and 11 had more variation, compared with BTV serotypes 13 and 17, especially for migration of genome segment 5. A definitive correlation was not seen between the double-stranded RNA migration profiles on polyacrylamide gel electrophoresis, geographic origin, herd of origin, or year of collection. One BTV field isolate contained more than 1 electropherotype, with 2 bands at the segment-7 position, and it was further characterized as BTV serotype 11. Segments 2 and 5 of EHDV isolates were more variable in their migration than were the other gene segments. Generally, migration profiles for EHDV double-stranded RNA were more variable, compared with those of BTV isolates. Although a correlation was found between migration profiles and serotype of 2 isolates of EHDV, a study of additional EHDV isolates is required before the diversity of electrophoretic patterns of EHDV can be determined.     

Experimental model of Border Disease Virus infection in lambs: comparative pathogenicity of pestiviruses isolated in France and Tunisia

Pestiviruses have been isolated from live sheep pox Tunisian vaccines. Vaccination with these vaccines caused outbreaks of Border Disease in Tunisia. In order to study more precisely the pathogenicity of these isolates, three groups of eight four month old lambs from a pestivirus-free flock were infected by the intratracheal route with a French strain (AV) and two Tunisian isolates (SN3G and Lot21). Clinical, hematological, immunological and virological parameters were evaluated. The three groups developed mild fever and leucopaenia by day 3 to 6 post infection (pi). The differences in the weight curves were not significant. Viruses were isolated from the peripheral blood buffy coat cells by day 4 to 9 pi. Antibodies were present on day 16 pi following infection by the French strain and on day 21 pi with the Tunisian isolates. The results demonstrated that SN3G and Lot21 are almost similar to the French strain used as the reference strain. In field conditions, they could induce economical losses in naive flocks, alone or in association with other pathogens.     

Additional considerations to the use of single‐step genomic predictions in a dominance setting

Recent publications indicate that single‐step models are suitable to estimate breeding values, dominance deviations and total genetic values with acceptable quality. Additive single‐step methods implicitly extend known number of allele information from genotyped to non‐genotyped animals. This theory is well derived in an additive setting. It was recently shown, at least empirically, that this basic strategy can be extended to dominance with reasonable prediction quality. Our study addressed two additional issues. It illustrated the theoretical basis for extension and validated genomic predictions to dominance based on single‐step genomic best linear unbiased prediction theory. This development was then extended to include inbreeding into dominance relationships, which is a currently not yet solved issue. Different parametrizations of dominance relationship matrices were proposed. Five dominance single‐step inverse matrices were tested and described as C¹ , C² , C³ , C⁴ and C⁵ . Genotypes were simulated for a real pig population (n = 11,943 animals). In order to avoid any confounding issues with additive effects, pseudo‐records including only dominance deviations and residuals were simulated. SNP effects of heterozygous genotypes were summed up to generate true dominance deviations. We added random noise to those values and used them as phenotypes. Accuracy was defined as correlation between true and predicted dominance deviations. We conducted five replicates and estimated accuracies in three sets: between all ( S₁ ), non‐genotyped ( S₂ ) and inbred non‐genotyped ( S₃ ) animals. Potential bias was assessed by regressing true dominance deviations on predicted values. Matrices accounting for inbreeding ( C³ , C⁴ and C⁵ ) best fit. Accuracies were on average 0.77, 0.40 and 0.46 in S₁ , S₂ and S₃ , respectively. In addition, C³ , C⁴ and C⁵ scenarios have shown better accuracies than C¹ and C² , and dominance deviations were less biased. Better matrix compatibility (accuracy and bias) was observed by re‐scaling diagonal elements to 1 minus the inbreeding coefficient ( C⁵ ).     

Regional and maturational effects on essential oils yields and composition of coriander (Coriandrum sativum L.) fruits

The composition of volatile components of the essential oils extracted from fruits of coriander (Coriandrum sativum L.) growing in two different Tunisian regions in both Menzel Temime and Borj El Ifaa was studied by GC and GC–MS. The highest oil yields were observed at final stages of maturity: 0.324 ± 0.09% and 0.327 ± 0.08%, in Menzel Temime and Borj El Ifaa, respectively. Essential oil yields were highly (P < 0.001) affected by the growing region, stages of maturity and their interaction. Essential oil composition changed significantly (P < 0.05) among the different stages of maturity and growing region. Linalool and geranyl acetate were the main compounds at full fruit maturity in the two studied regions. Growing region, stages of maturity and their interaction had a strong effect (P < 0.001) on 35 compounds. α-Pinene, sabinene, limonene, γ-terpinene, cis-dihydrocarvone and geranial were insensible to the regional factor.     

Complete sequencing of Tunisian redspotted grouper nervous necrosis virus betanodavirus capsid gene and RNA‐dependent RNA polymerase gene

Finfish nodaviruses (betanodaviruses) can cause highly destructive infections in numerous species of farmed marine fish larvae and juveniles worldwide. The betanodavirus genome consists of two single‐stranded positive‐sense RNA molecules (RNA1 and RNA2). The virus can be classified into four genotypes based on the partial sequences of the coat protein (CP) gene (T2 and T4 regions). Currently, genomic sequence information for RNA1 regions of RNA2 outside of T2 and T4 is less well documented. This study reports on the characterization of the full RNA2 sequence of a Tunisian betanodavirus with a length of 1433 nt, containing a 339 amino acid open‐reading frame encoding the CP, and typing to the redspotted grouper nervous necrosis virus Ia genotype following phylogenetic analysis. The homology of the capsid protein to other betanodaviruses or alphanodaviruses was compared. In addition, a full length RNA1 sequence of 3104 nt encoding a 982 amino acid RNA‐dependent RNA polymerase was obtained.     

Vaccination of Tunisian dogs with the lyophilised SAG2 oral rabies vaccine incorporated into the DBL2 dog bait

The protective effect of the lyophilised SAG2 oral vaccine bait DBL2, already demonstrated on laboratory dogs, needed to be verified on common Tunisian dogs. Seven Tunisian dogs consumed totally or partially one DBL2 bait containing 10(8.3) TCID50 of the highly attenuated rabies vaccine strain, SAG2. Five of the seven vaccinated animals survived a challenge administered 33 days later with a Tunisian canine street rabies virus to which five of the six controls that were not vaccinated and had no specific antibodies succumbed. The partial or total consumption of a single DBL2 bait thus conferred a protective immune response similar to that observed in laboratory dogs to dogs of poor health status. The sero-antibody response was, however, weak: only two vaccinated dogs exhibited a significant neutralising antibody response after vaccination and before the challenge, and four after the challenge.