REP Groups Research Articles

Short- and mid-term effects on performance, health and qualitative behavioural assessment of Romane lambs in different milk feeding conditions

The common practice of artificially rearing lambs from prolific meat breeds of sheep constitutes a welfare issue due to increased mortality rates and negative health issues. In this multidisciplinary study, we investigated the possible short- and mid-term advantages of artificially feeding fresh ewe's milk instead of commercial milk replacer on lambs' growth, health and welfare. Romane lambs were either separated from their mothers on D3 and fed with Lacaune ewes' milk (LAC, n = 13) or milk replacer (REP, n = 15), or they were reared by their mothers (MOT, n = 15). On D45, they were weaned, gathered in single-sex groups until the end of the study on D150. Lamb performance and biomarkers of overall health were assessed by measuring: growth, dirtiness of the perianal area, enteric pathogens in the faeces, total antioxidant status and redox status assessed by plasma reduced glutathione/oxidised glutathione ratio, and immune response after vaccination against chlamydiosis. As an exploratory approach, blood cell transcriptomic profiles were also investigated. Last, qualitative behaviour assessment (QBA) was performed as an integrated welfare criterion. Lacaune ewes' milk and REP never differed in their average daily gain but grew less than MOT lambs in the early suckling period and just after weaning. No effect was detected afterwards. On D30, LAC and REP lambs had lower total antioxidant and higher redox status than MOT lambs but did not differ among themselves. Lacaune ewes' milk and MOT had a cleaner perianal area than REP lambs on D21, while faecal pathogen infection did not vary between the treatment groups. After vaccination, LAC also had a stronger immune response on D90 compared to REP lambs. Transcriptome analysis performed on D150 showed differential gene expression, mainly in relation to inflammatory, immune and cell cycle response, between male lambs of the LAC group and those of the MOT and REP groups. Based on QBA, LAC lambs never differed from MOT lambs in their general activity and varied from REP only on D21; REP lambs were always more agitated than MOT lambs. In conclusion, artificial milk feeding impaired early growth rate, health and emotional state mainly during the milk feeding period and at weaning. Feeding artificially reared lambs with fresh ewe's milk partly mitigated some of the negative effects induced by milk replacer but without achieving the full benefit of being reared by the mother.

Analysis of Dextromethorphan and Three Metabolites in Decomposed Skeletal Tissues by UPLC-QToF-MS: Comparison of Acute and Repeated Drug Exposures.

Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS) analysis of dextromethorphan (DXM) and its metabolites-dextrorphan, 3-methoxymorphinan (3-MEM) and 3-hydroxymorphinan-in skeletal remains of rats exposed to DXM under different dosing patterns is described. Rats (n=20) received DXM in one of four dosing patterns: acute (ACU1 or ACU2-100 or 200mg/kg, i.p.; n=5, respectively) or repeated (REP1 or REP2-3 doses of 25 or 50mg/kg, i.p., 30min apart; n=5, respectively). Drug-free animals (n=5) served as negative controls. Following euthanasia, the animals decomposed to skeleton outdoors. Bones were sorted by animal and skeletal element (vertebra, femur, pelvis, tibia, rib and skull), washed, air-dried and pulverized prior to dynamic methanolic drug extraction, filtration/pass-through extraction and analysis by UPLC-QToF-MS in positive electrospray ionization mode. Analyte levels (expressed as mass-normalized response ratios, RR/m) differed significantly between ACU1 and ACU2 (Mann-Whitney (MW), P<0.05) in all skeletal elements for all analytes investigated, and between REP1 and REP2 in most skeletal elements for 3-MEM and 3-HOM, but in all skeletal elements for DXM. Between ACU1 and ACU2, and between REP1 and REP2, analyte level ratios (RRi/RRj) differed significantly (MW, P<0.05) in 3/6 to 6/6 skeletal elements, depending on the ratios concerned, with no analyte level ratio differing significantly between both ACU1 vs ACU2 and REP1 vs REP2. Kruskal-Wallis (KW) analysis showed skeletal element to be a main effect for all analyte levels and analyte level ratios in all ACU and REP groups examined (P<0.05). For data pooled only according to exposure pattern, KW analysis showed dose pattern to be a main effect for both analyte levels and analyte level ratios (P<0.05). These data illustrate a dependence of these measures on dose, dose pattern and skeletal element, suggesting that some exposure patterns may be distinguished by toxicological analysis of bone.

Are exercise professionals fit to provide nutrition advice? An evaluation of general nutrition knowledge

ObjectivesRegistered exercise professionals (REP) are trained to provide structured exercise recommendations and general nutrition advice to healthy clients. However REP provide specific nutrition advice beyond their scope of practice, including diet-disease advice. The present study aims to investigate the level of general nutrition knowledge of REP, and compare this to a sample of community members (CTM), and university trained dietitians (DN). DesignAge-matched REP, CTM and DN were recruited to complete the previously validated revised-general nutrition knowledge questionnaire. MethodsTotal nutrition knowledge score and section scores were compared between REP, CTM and DN. The impact of sex, age, level of education, and years’ experience on nutrition knowledge was investigated. ResultsA total of 554 participants completed the questionnaire (REP, n=161; CTM, n=357; DN, n=36). The DN group performed significantly better overall (DN, 91.2±4.6; REP, 78.4±9.6; CTM, 75.4±11.3) and for knowledge of dietary guidelines (DN, 82.3±20.7; REP, 80.5±15.5; CTM, 80.0±14.1), nutrient content of foods (DN, 92.6±4.4; REP, 80.9±9.9; CTM, 75.7±12.0), and diet-disease relationships (DN, 91.4±9.7; REP, 65.4±18.0; CTM, 68.6±11.1) compared to the REP and CTM groups (p<0.001). Sex, education and age were all significant predictors of total nutrition knowledge (p<0.0005). ConclusionsTotal nutrition knowledge and knowledge of diet-disease relationships is limited in REP. Encouraging REP to work collaboratively with a multidisciplinary team, including DN, will assist in providing optimal client care in achieving health and body composition related goals.

Pervasive Chimerism in the Replication-Associated Proteins of Uncultured Single-Stranded DNA Viruses.

Numerous metagenomic studies have uncovered a remarkable diversity of circular replication-associated protein (Rep)-encoding single-stranded (CRESS) DNA viruses, the majority of which are uncultured and unclassified. Unlike capsid proteins, the Reps show significant similarity across different groups of CRESS DNA viruses and have conserved domain organization with the N-terminal nuclease and the C-terminal helicase domain. Consequently, Rep is widely used as a marker for identification, classification and assessment of the diversity of CRESS DNA viruses. However, it has been shown that in certain viruses the Rep nuclease and helicase domains display incongruent evolutionary histories. Here, we systematically evaluated the co-evolutionary patterns of the two Rep domains across classified and unclassified CRESS DNA viruses. Our analysis indicates that the Reps encoded by members of the families Bacilladnaviridae, Circoviridae, Geminiviridae, Genomoviridae, Nanoviridae and Smacoviridae display largely congruent evolutionary patterns in the two domains. By contrast, among the unclassified CRESS DNA viruses, 71% appear to have chimeric Reps. Such massive chimerism suggests that unclassified CRESS DNA viruses represent a dynamic population in which exchange of gene fragments encoding the nuclease and helicase domains is extremely common. Furthermore, purging of the chimeric sequences uncovered six monophyletic Rep groups that may represent new families of CRESS DNA viruses.

Isolation and molecular characterization of genetically diverse antagonistic, diazotrophic red-pigmented vibrios from different mangrove rhizospheres

Genetic diversity of red-pigmented vibrios from different mangrove rhizospheres (Porteresia coarctata, Avicennia marina and Rhizophora mucronata) collected from Pichavaram mangrove, India was investigated. Twenty red-pink pigmented strains were isolated, 16S rRNA gene analyses indicted that these isolates belong to the genus Vibrio and were phylogenetically closely related to the type strains of Vibrio rhizosphaerae and Vibrio ruber. The rep-PCR analysis using GTG(5) and BOX primers had similar groupings, and segregated these pigmented Vibrio isolates including two type strains into seven unique genotypic groups (REP groups A1-A7). The rhizosphere of P. coarctata harbors highly genetically diverse groups of red-pigmented vibrios compared to other plants. Multilocus sequence analysis using four genetic loci (pyrH, recA, rpoA, 16S rRNA) clearly supported the hypothesis that strains MSSRF38 (REP group A5) and MSSRF39 (REP group A6) could represent new Vibrio species. Biological functions of these vibrios were also determined and it was found that all these isolates have antagonistic activity against phytopathogens, and isolates belonging to REP groups A5 and A6 were positive for nifH gene by PCR. In conclusion, this study for the first time demonstrates the occurrence of genetically diverse groups of antagonistic, diazotrophic red-pigmented vibrios from different mangrove plants and suggests a new ecological role for vibrios as heterotrophic plant associated rhizobacteria.

CA‐125 levels in ruptured and unruptured tubal ectopic pregnancies

This prospective, single-blind and controlled clinical study aimed to research if CA-125 levels could be a useful test in the differential diagnosis of intact and ruptured tubal ectopic pregnancy. Sixty-five women with tubal ectopic pregnancy of 5-10 weeks' duration (27 women with ruptured tubal ectopic pregnancy [REP] and 38 women with unruptured tubal ectopic pregnancy [UREP]) and 65 women with normal intrauterine pregnancy (NIUP) of the same gestational age were studied prospectively. Serum CA-125 levels were measured in all women and these levels were compared among the REP, UREP, and NIUP groups. The mean CA-125 levels didn't show any significant difference between the REP and NIUP groups (P > 0.05). The mean CA-125 levels of these two groups were higher than that in the UREP group (P < 0.01, P < 0.001, respectively). The dispersion ratios of the CA-125 levels had a statistically significant difference between the REP and UREP groups (chi(2): 42.44, P < 0.0001). CA-125 levels weren't correlated with gestational weeks in the REP and UREP groups (r: 0.005, P > 0.05; r: 0.008, P > 0.05, respectively). In intact tubal ectopic pregnancies, expectant or managed with medical treatment, the increase of CA-125 levels in the serial measurements could be a supplementary test for an early diagnosis of tubal rupture.

Muscular adaptations in response to three different resistance-training regimens: specificity of repetition maximum training zones.

Thirty-two untrained men [mean (SD) age 22.5 (5.8) years, height 178.3 (7.2) cm, body mass 77.8 (11.9) kg] participated in an 8-week progressive resistance-training program to investigate the "strength-endurance continuum". Subjects were divided into four groups: a low repetition group (Low Rep, n = 9) performing 3-5 repetitions maximum (RM) for four sets of each exercise with 3 min rest between sets and exercises, an intermediate repetition group (Int Rep, n = 11) performing 9-11 RM for three sets with 2 min rest, a high repetition group (High Rep, n = 7) performing 20-28 RM for two sets with 1 min rest, and a non-exercising control group (Con, n = 5). Three exercises (leg press, squat, and knee extension) were performed 2 days/week for the first 4 weeks and 3 days/week for the final 4 weeks. Maximal strength [one repetition maximum, 1RM), local muscular endurance (maximal number of repetitions performed with 60% of 1RM), and various cardiorespiratory parameters (e.g., maximum oxygen consumption, pulmonary ventilation, maximal aerobic power, time to exhaustion) were assessed at the beginning and end of the study. In addition, pre- and post-training muscle biopsy samples were analyzed for fiber-type composition, cross-sectional area, myosin heavy chain (MHC) content, and capillarization. Maximal strength improved significantly more for the Low Rep group compared to the other training groups, and the maximal number of repetitions at 60% 1RM improved the most for the High Rep group. In addition, maximal aerobic power and time to exhaustion significantly increased at the end of the study for only the High Rep group. All three major fiber types (types I, IIA, and IIB) hypertrophied for the Low Rep and Int Rep groups, whereas no significant increases were demonstrated for either the High Rep or Con groups. However, the percentage of type IIB fibers decreased, with a concomitant increase in IIAB fibers for all three resistance-trained groups. These fiber-type conversions were supported by a significant decrease in MHCIIb accompanied by a significant increase in MHCIIa. No significant changes in fiber-type composition were found in the control samples. Although all three training regimens resulted in similar fiber-type transformations (IIB to IIA), the low to intermediate repetition resistance-training programs induced a greater hypertrophic effect compared to the high repetition regimen. The High Rep group, however, appeared better adapted for submaximal, prolonged contractions, with significant increases after training in aerobic power and time to exhaustion. Thus, low and intermediate RM training appears to induce similar muscular adaptations, at least after short-term training in previously untrained subjects. Overall, however, these data demonstrate that both physical performance and the associated physiological adaptations are linked to the intensity and number of repetitions performed, and thus lend support to the "strength-endurance continuum".

Taxonomic characterization of Ochrobactrum sp. isolates from soil samples and wheat roots, and description of Ochrobactrum tritici sp. nov. and Ochrobactrum grignonense sp. nov.

A large collection of bacterial strains, immunotrapped from soil and from the wheat rhizoplane, was subjected to polyphasic taxonomy by examining various pheno- and genotypic parameters. Strains were grouped on (inter) repetitive extragenic palindromic DNA (REP) PCR profiles at the intraspecies level. Pheno- and genotypic characters were assessed for representatives from 13 different REP groups. Strains of nine REP groups constituting two physiological BIOLOG clusters fell in the coherent DNA-DNA reassociation group of Ochrobactrum anthropi. Strains of two REP groups constituting a separate BIOLOG cluster fell in the coherent DNA-DNA reassociation group of Ochrobactrum intermedium. Additional phenotypic characters differentiating O. anthropi and O. intermedium were found. REP group K strains constituted a different BIOLOG cluster, a separate DNA-DNA reassociation group and a distinct phylogenetic lineage in 165 rDNA homology analysis, indicating that REP group K strains represent a new species. Diagnostic phenotypic characters were found. Closest relatives were Ochrobactrum species. The name Ochrobactrum grignonense sp. nov. is proposed (type strain OgA9aT = LMG 18954T = DSM 13338T). REP group J strains again constituted a different BIOLOG cluster, a separate DNA-DNA reassociation group and showed, as a biological particularity, a strict preference for the rhizoplane as habitat. Diagnostic phenotypic characters were found. This indicated that REP group J strains represent a further new species, although phylogenetic analyses using 16S rDNA homology were not able to separate the cluster of REP group J sequences significantly from 16S rDNA sequences of Ochrobactrum anthropi. The name Ochrobactrum tritici sp. nov. is proposed (type strain SCII24T = LMG 18957T = DSM 13340T).

Genetic Diversity Among Rhizobial Isolates from Field-grown Astragalus sinicus of Southern China

Summary The genetic diversity among 204 rhizobial isolates from field-grown Astragalus sinicus collected from 52 ecological different sites of seven southern provinces of China was investigated by using rep-PCR fingerprinting with REP primers and 16S rDNA PCR-RFLP and sequence analyses. Most isolates (78.4%) belonged to different 16S rDNA genotypes from that of Mesorhizobium huakuii type strain CCBAU2609 T . Rep-PCR fingerprinting analysis of 136 randomly chosen isolates categorized 27 REP groups at a similarity level of 60%. Genetically diverse rhizobial strains could be detected at single site, and strains with the same genetic type could be found in different geographic locations. Twelve representative strains were selected for identification using 16S and 23S rDNA analyses, which resulted in four 16S and five 23S rDNA PCR-RFLP genotypes. When both 16S and 23S rDNA PCR-RFLP data were combined together, seven different genotypes were detected among the 12 representative strains, and no strain shared a genotype with the M. huakuii type strain. Both 16S and 23S rRNA genes analyses clustered the 12 representative stains within the Mesorhizobium branch. The 16S rRNA results obtained by PCR-RFLP were confirmed by sequencing the first 600 bp of the 16S rRNA genes.

Genetic diversity of rhizobial symbionts isolated from legume species within the genera Astragalus, Oxytropis, and Onobrychis.

The genetic diversity of 44 rhizobial isolates from Astragalus, Oxytropis, and Onobrychis spp. originating from different geographic locations was evaluated by mapped restriction site polymorphism (MRSP) analysis of 16S rRNA genes and by PCR DNA fingerprinting with repetitive sequences (REP-PCR). A comparison of tree topologies of reference strains constructed with data obtained by MRSP and by 16S rRNA gene sequence analyses showed that the topologies were in good agreement, indicating that the MSRP approach results in reasonable estimates of rhizobial phylogeny. The isolates were distributed into 14 distinct 16S rRNA gene types clustering into three major groups which corresponded with three of the genera within the legume symbionts. Most of the isolates were within the genus Mesorhizobium. Five were identified with different genomic species nodulating Lotus spp. and Cicer arietinum. Three Astragalus isolates were classified as Bradyrhizobium, one being similar to Bradyrhizobium elkanii and another being similar to Bradyrhizobium japonicum. Six of the isolates were related to species within the genus Rhizobium. Two were similar to Rhizobium leguminosarum, and the remainder were identified as Rhizobium gallicum. DNA fingerprinting by REP-PCR revealed a high level of diversity within single 16S ribosomal DNA types. The 44 isolates were distributed into 34 REP groups. Rhizobial classification at the genus and probably also the species levels was independent of geographic origin and host plant affinity.

Discrete-trial probability learning in rats: Effects of local contingencies of reinforcement

Three groups of rats were trained using a noncorrection procedure on a probability learning task in which responses on one lever produced reinforcement on 61% of the trials and responses on the other lever produced reinforcement on the remaining trials. For Group RAN, there were no sequential constraints, but for Groups REP and ALT, the lever on which reinforcement was produced on a given trial depended upon the lever that had been designated correct on the previous trial. For Group REP, the locus of reinforcement was repeated on approximately 80% of trials; Group ALT was required to switch to the other lever on approximately 80% of trials. The maximizing strategy for Group RAN was exclusive responding to the majority lever, for Group ALT, it was “win-shift, lose-stay,” and for Group REP, “win-stay, lose-shift.” Subjects from both the REP and ALT groups tended to maximize momentarily, thereby showing a lower relative frequency of majority responses than did subjects from the RAN group. The differences between groups cannot be accounted for by a molar analysis; a molecular analysis is needed.