Unravelling the secrets of non-host resistance in plant-insect interactions.

Despite the existence of over half million species of plant-eating insects, our planet remains predominantly green. In fact, susceptibility to herbivory is the exception, as plants are resistant to most insect species. This phenomenon is known as nonhost resistance (NHR), where every individual of a plant species is resistant to all variants of a pest or pathogen. While NHR represents the most common and durable outcome of the plant immune system, several aspects of this type of plant defence remains elusive, particularly in plant-insect interactions. In this review, we clarify the concepts of NHR in plant-insect interaction. We emphasize that NHR is a phenomenon arising as a consequence of effective plant defences providing invulnerability to most insect herbivores. This underscores that NHR is one of the main ecological features delimiting the range of plant-insect interactions on Earth. We further highlight the traits and molecular components of the plant immune system known to participate in NHR against insects. Finally, we discuss how NHR can be leveraged as a tool to develop pest resilient crops. Given the significant threat insects pose to global food security, research in plant NHR represents a crucial focal point with immense potential for ensuring food security worldwide.

Screening of CIMMYT and South Asian Bread Wheat Germplasm Reveals Marker-Trait Associations for Seedling Resistance to Septoria Nodorum Blotch.

Wheat (Triticum aestivum L.) production is adversely impacted by Septoria nodorum blotch (SNB), a fungal disease caused by Parastagonospora nodorum. Wheat breeders are constantly up against this biotic challenge as they try to create resistant cultivars. The genome-wide association study (GWAS) has become an efficient tool for identifying molecular markers linked with SNB resistance. This technique is used to acquire an understanding of the genetic basis of resistance and to facilitate marker-assisted selection. In the current study, a total of 174 bread wheat accessions from South Asia and CIMMYT were assessed for SNB reactions at the seedling stage in three greenhouse experiments at CIMMYT, Mexico. The results indicated that 129 genotypes were resistant to SNB, 39 were moderately resistant, and only 6 were moderately susceptible. The Genotyping Illumina Infinium 15K Bead Chip was used, and 11,184 SNP markers were utilized to identify marker-trait associations (MTAs) after filtering. Multiple tests confirmed the existence of significant MTAs on chromosomes 5B, 5A, and 3D, and the ones at Tsn1 on 5B were the most stable and conferred the highest phenotypic variation. The resistant genotypes identified in this study could be cultivated in South Asian countries as a preventative measure against the spread of SNB. This work also identified molecular markers of SNB resistance that could be used in future wheat breeding projects.

Comparative analysis of Rhipicephalus microplus (Canestrini, 1888) infestation in pure breed (Hereford) and cross breed (Braford) cattle herds subjected to the same chemical treatments.

The aim of this work was to analyze the R. microplus (Canestrini, 1888) infestation in two bovine herds with different degrees of natural resistance (i.e., Hereford and Braford) to ticks subjected to an identical chemical treatment scheme to ticks at the same farm, to demonstrate the impact on tick control of the incorporation of a more resistant bovine breed. Two groups of ten Hereford and Braford cows each were subjected to eleven chemical treatments between August 2022 and October 2023 (four fluazuron, two fipronil 1%, one ivermectin 3.15% and four immersion in a dipping vat with a combination of cypermethrin 10% and ethion 40%). Tick population was shown to be susceptible to ivermectin, fluazuron and the mix cypermethrin 10%-ethion 40% and resistant to fipronil according to in vitro tests. Tick infestation was significantly greater in the Hereford cows than in the Braford cows. Tick infestation in both Hereford and Braford breeds was similar when treatment with functional drugs was applied, but when a block of the treatments was done with drugs with decreased functionality due to resistance (i.e. fipronil), treatment failure was manifested more strongly in the most susceptible breed. The incorporation of cattle breeds with moderate or high resistance to R. microplus is instrumental to optimize the efficacy and sustainability of chemical control of ticks in a scenario where resistance to one or more chemical groups is almost ubiquitous, because it favors the biological control of this parasite.

Molecular Interplay between Non-Host Resistance, Pathogens and Basal Immunity as a Background for Fatal Yellowing in Oil Palm (Elaeis guineensis Jacq.) Plants.

An oil palm (Elaeis guineensis Jacq.) bud rod disorder of unknown etiology, named Fatal Yellowing (FY) disease, is regarded as one of the top constraints with respect to the growth of the palm oil industry in Brazil. FY etiology has been a challenge embraced by several research groups in plant pathology throughout the last 50 years in Brazil, with no success in completing Koch's postulates. Most recently, the hypothesis of having an abiotic stressor as the initial cause of FY has gained ground, and oxygen deficiency (hypoxia) damaging the root system has become a candidate for stress. Here, a comprehensive, large-scale, single- and multi-omics integration analysis of the metabolome and transcriptome profiles on the leaves of oil palm plants contrasting in terms of FY symptomatology-asymptomatic and symptomatic-and collected in two distinct seasons-dry and rainy-is reported. The changes observed in the physicochemical attributes of the soil and the chemical attributes and metabolome profiles of the leaves did not allow the discrimination of plants which were asymptomatic or symptomatic for this disease, not even in the rainy season, when the soil became waterlogged. However, the multi-omics integration analysis of enzymes and metabolites differentially expressed in asymptomatic and/or symptomatic plants in the rainy season compared to the dry season allowed the identification of the metabolic pathways most affected by the changes in the environment, opening an opportunity for additional characterization of the role of hypoxia in FY symptom intensification. Finally, the initial analysis of a set of 56 proteins/genes differentially expressed in symptomatic plants compared to the asymptomatic ones, independent of the season, has presented pieces of evidence suggesting that breaks in the non-host resistance to non-adapted pathogens and the basal immunity to adapted pathogens, caused by the anaerobic conditions experienced by the plants, might be linked to the onset of this disease. This set of genes might offer the opportunity to develop biomarkers for selecting oil palm plants resistant to this disease and to help pave the way to employing strategies to keep the safety barriers raised and strong.

Phenotypic evaluation of genetic resistance to the tick Rhipicephalus (Boophilus) microplus in Argentine Creole cattle.

The objective of this work was to characterize the Argentine Creole cattle breed through the identification of individual phenotypic variations in the levels of infestation with Rhipicephalus (Boophilus) microplus. We evaluated 179 heifers exposed to successive artificial infestations from 2015 to 2018, achieving a total of 663 observations. Tick counts were assessed with the linear mixed model, considering year of evaluation, time of infestation, dam's age and nutritional status during the evaluated period as fixed effects. The average tick count value obtained allowed to classify the breed as highly resistant to the tick charge (99.3%). Although the previous nutritional condition of the animals did not affect the individual charge response, weight gain during the trial showed a significantly negative correlation. We conclude that the Argentine Creole breed is an attractive genetic alternative for cattle breeding in endemic regions, either as a pure breed or a cross-breed.

The potential of PGPR and Trichoderma-based bioproducts and resistant cultivars as tools to manage clubroot disease in cruciferous crops.

The objective of this research was to determine the potential use of eco-friendly technologies to reduce the clubroot disease caused by Plasmodiophora brassicae, the main constraint of cruciferous crops worldwide. Two commercial bioproducts were evaluated in susceptible broccoli, one based on the PGPR consortium (Bacillus amyloliquefaciens, Bacillus pumilus, and Agrobacterium radiobacter K84) and the other one based on Trichoderma koningiopsis Th003 (Tricotec® WG). Additionally, the resistant broccoli cv. Monclano® was tested under two concentrations of resting spores (RS) of P. brassicae, 1 × 103 and 1 × 105 RS g-1 of soil. The first phase of evaluations with broccoli was carried out under a greenhouse, while susceptible broccoli, cauliflower, and red cabbage were included in a subsequent field phase. Tebuconazole + Trifloxystrobin mixture and Fluazinam were included as positive controls. The effectiveness of the bioproducts depended on the nature of the biocontrol agent, the concentration of P. brassicae, and the dose of treatment. Tricotec® showed consistent plant growth promotion but no biocontrol effect against clubroot, and the rhizobacteria-based bioproduct significantly reduced the disease in both greenhouse and field experiments. Higher disease severity was observed with the higher dose of Tricotec®. Under field conditions, the rhizobacteria reduced the incidence progress by 26%, 39%, and 57% under high, medium, and low pressure of the pathogen, respectively. However, no reduction of clubroot severity under high pressure of the pathogen was observed. Complete inhibition of club formation in roots was achieved via the fungicide, but a phytotoxic effect was observed under greenhouse conditions. Fungicides reduced the incidence progress of clubroot, but not the severity under high inoculum pressure in the field. The fungicides, the bacterial treatment, and the combination of bioproducts tended to delay the progress of the disease compared with the negative control and Tricotec alone. The resistant broccoli showed a low level of disease under high concentrations of P. brassicae (less than 10% incidence and up to 2% severity). These results suggested the overall potential of commercial tools based on the PGPR consortium and plant resistance to control P. brassicae. The integration of control measures, the role of Trichoderma spp. in P. brassicae-cruciferous pathosystems, and the need to recover highly infested soils will be discussed.

Impact of the Paper by Allen and Humphreys (1979) on Anti-Tick Vaccine Research.

The classic paper by Allen and Humphreys "Immunisation of guinea pigs and cattle against ticks" Nature, 1979, 280: 491-493 led to a surge in the development of tick vaccines as a nonchemical method for prevention of tick infestations in susceptible hosts living in tick-endemic regions. Although observations of host resistance to ticks had been documented since the beginning of the last century, it was not until publication of this paper that the proof of concept of anti-tick vaccines was developed. The described experimental methods directly impacted further investigations on the discovery and evaluation of new anti-tick vaccines.

Host Serum Proteins as Potential Biomarkers of Bovine Tuberculosis Resistance Phenotype.

Eradication of bovine tuberculosis (bTB) continues to be a worldwide challenge. The lack of reliable vaccines dampens the control and eradication programs of Mycobacterium bovis infection and spread. Selection and breeding of cattle resistant to M. bovis infection would greatly enhance the effectiveness of bTB eradication programs. Here, we have evaluated the potential of serum proteins as biomarkers of cattle resistance to bTB in Holstein-Friesian cows, 6-8-year-old, born and raised in similar conditions in herds with bTB prevalence >30%. Serum proteins obtained from uninfected cows (bTB-resistant; R) were compared to those from infected cows (bTB-susceptible; S), defined by a negative or positive bTB diagnosis, respectively. bTB diagnosis included: (i) single intradermal (caudal fold) tuberculin test, (ii) whole blood IFN-gamma test, (iii) gross visible lesions in lymph nodes and lungs by inspection at the abattoir, and (iv) a bacteriological culture for M. bovis. Using 2D-GE and LC-ESI-MS/MS, we found higher expression levels of primary amine oxidase (AO), complement component 5 (C5), and serotransferrin (TF) in R cattle than S cattle. In-house developed and standardized ELISAs for these novel biomarkers showed the best sensitivities of 72, 77, 77%, and specificities of 94, 94, 83%, for AO, C5, and TF, respectively. AUC-ROC (95% CI) values of 0.8935 (0.7906-0.9964), 0.9290 (0.8484-1.010), and 0.8580 (0.7291-0.9869) were obtained at cut-off points of 192.0, 176.5 ng/ml, and 2.1 mg/ml for AO, C5, and TF, respectively. These proteins are involved in inflammatory/immunomodulatory responses to infections and may provide a novel avenue of research to determine the mechanisms of protection against bTB. Overall, our results indicate that these proteins could be novel biomarkers to help identify cattle resistant to bTB, which in turn could be used to strengthen the effectiveness of existing eradication programs against bTB.

Host Susceptibility Modulates Escovopsis Pathogenic Potential in the Fungiculture of Higher Attine Ants.

Health and disease emerge from intricate interactions between genotypes, phenotypes, and environmental features. The outcomes of such interactions are context-dependent, existing as a dynamic continuum ranging from benefits to damage. In host-microbial interactions, both the host and environmental conditions modulate the pathogenic potential of a microorganism. Microbial interactions are the core of the agricultural systems of ants in the subtribe Attina, which cultivate basidiomycete fungi for food. The fungiculture environment harbors a diverse microbial community, including fungi in the genus Escovopsis that has been studied as damage-causing agent. Here, we consider the ant colony as a host and investigate to what extent its health impacts the dynamics and outcomes of host-Escovopsis interactions. We found that different ant fungal cultivars vary in susceptibility to the same Escovopsis strains in plate-assays interactions. In subcolony-Escovopsis interactions, while healthy subcolonies gradually recover from infection with different concentrations of Escovopsis conidia, insecticide-treated subcolonies evidenced traits of infection and died within 7 days. The opportunistic nature of Escovopsis infections indicates that diseases in attine fungiculture are a consequence of host susceptibility, rather than the effect of a single microbial agent. By addressing the host susceptibility as a major modulator of Escovopsis pathogenesis, our findings expand the understanding of disease dynamics within attine colonies.

Multiple Country and Breed Genomic Prediction of Tick Resistance in Beef Cattle.

Ticks cause substantial production losses for beef and dairy cattle. Cattle resistance to ticks is one of the most important factors affecting tick control, but largely neglected due to the challenge of phenotyping. In this study, we evaluate the pooling of tick resistance phenotyped reference populations from multi-country beef cattle breeds to assess the possibility of improving host resistance through multi-trait genomic selection. Data consisted of tick counts or scores assessing the number of female ticks at least 4.5 mm length and derived from seven populations, with breed, country, number of records and genotyped/phenotyped animals being respectively: Angus (AN), Brazil, 2,263, 921/1,156, Hereford (HH), Brazil, 6,615, 1,910/2,802, Brangus (BN), Brazil, 2,441, 851/851, Braford (BO), Brazil, 9,523, 3,062/4,095, Tropical Composite (TC), Australia, 229, 229/229, Brahman (BR), Australia, 675, 675/675, and Nguni (NG), South Africa, 490, 490/490. All populations were genotyped using medium density Illumina SNP BeadChips and imputed to a common high-density panel of 332,468 markers. The mean linkage disequilibrium (LD) between adjacent SNPs varied from 0.24 to 0.37 across populations and so was sufficient to allow genomic breeding values (GEBV) prediction. Correlations of LD phase between breeds were higher between composites and their founder breeds (0.81 to 0.95) and lower between NG and the other breeds (0.27 and 0.35). There was wide range of estimated heritability (0.05 and 0.42) and genetic correlation (-0.01 and 0.87) for tick resistance across the studied populations, with the largest genetic correlation observed between BN and BO. Predictive ability was improved under the old-young validation for three of the seven populations using a multi-trait approach compared to a single trait within-population prediction, while whole and partial data GEBV correlations increased in all cases, with relative improvements ranging from 3% for BO to 64% for TC. Moreover, the multi-trait analysis was useful to correct typical over-dispersion of the GEBV. Results from this study indicate that a joint genomic evaluation of AN, HH, BN, BO and BR can be readily implemented to improve tick resistance of these populations using selection on GEBV. For NG and TC additional phenotyping will be required to obtain accurate GEBV.

Local and systemic immune mediators of Morada Nova lambs with divergent Haemonchus contortus resistance phenotypes.

AIMS: Local and systemic immune mediators of Morada Nova lambs with divergent Haemonchus contortus resistance phenotypes were evaluated. METHODS AND RESULTS: Lambs were ranked through faecal egg counts (FEC) after two parasitic challenges with 4,000 H.contortus L3 . After the second challenge, the lambs underwent a third artificial infection and were euthanized 7 days later. Immune-related genes were quantified locally in abomasal mucosa and lymph nodes (CD4, IFNγ, IL4, IL5, IL13, IL2RA and MS4A2) and systemically in the whole blood (IL4 and IL13). Anti-H. contortus IgG and IgA antibodies and eosinophils and mast cells counts were also investigated. Resistant animals presented higher systemic IgG and IgA titres, both negatively correlated with FEC. Susceptible animals had higher blood levels of IL4 transcripts. At the local level, resistant lambs had higher eosinophils counts and superior MS4A2 levels in abomasal fundic mucosa, besides higher IgA levels in abomasal mucus, while susceptible lamb had superior IL4 expression in abomasal lymph nodes. CONCLUSION: These data indicate that resistant lambs had an immune response mediated by antibody-mediated cytotoxicity. Also, the systemic humoral profile, particularly IgA isotype, seems to be a good resistance marker for Morada Nova sheep, as we found differences between groups even when FEC did not differ.

Four single nucleotide polymorphisms (SNPs) are associated with resistance and resilience to Haemonchus contortus in Brazilian Morada Nova sheep.

Gastrointestinal nematodes are a major constraint in sheep production. Breeding for resistance has proven to be an effective and feasible approach to address this problem. The use and investigation of genetic markers for resistance traits could accelerate genetic progress and lead to a better understanding of underlying molecular mechanisms. Thus, the aim of this study was to evaluate if five single nucleotide polymorphisms SNPs OAR2_14765360, OAR6_81718546, OAR11_62887032, OAR12_69606944 and OAR15_59871543 are associated with resistance and resilience traits in a flock of the Morada Nova sheep breed. Lambs were submitted to two consecutive parasite challenges by oral infection with 4000 infective larvae L3) of Haemonchus contortus. Fecal egg counts (FEC), packed cell volume (PVC) and body weight were measured every one or two weeks for 42 days in each trial. DNA samples from 287 lambs, 131 ewes and 4 rams were amplified by ARMS-PCR or PCR-RFLP and genotypes were determined. Analysis of variance (ANOVA) was used for association analyses between genotypes and phenotypes. In case of significant association, the allele substitution effect was calculated based on a linear model. OAR2_14765360 and OAR12_69606944 were associated with FEC, and OAR12_69606944 also had significant effects on PCV and weight gain, showing favourable associations of the CC genotype with all evaluated traits. Both OAR6_81718546 and OAR11_62887032 were associated with weight gain, and OAR6_81718546 had an additional effect on PCV. OAR15_59871543 was not polymorphic in the population. OAR6_81718546 and OAR12_69606944 presented significant allele substitution effects of -1.06 ±â€¯0.52 kg for the T allele on final body weight and 0.74 ±â€¯0.32 for the C allele in PCV of the same sampling date, respectively. This is the first report of SNPs associated with gastrointestinal nematode resistance in this sheep breed. Our findings support the existence of quantitatice trait loci (QTL) for resistance and resilience in linkage disequilibrium with the polymorphic SNPs and suggest their future use for explorations of these traits in Morada Nova sheep.

Differential Accumulation of Innate- and Adaptive-Immune-Response-Derived Transcripts during Antagonism between Papaya Ringspot Virus and Papaya Mosaic Virus.

Papaya ringspot virus (PRSV), a common potyvirus infecting papaya plants worldwide, can lead to either antagonism or synergism in mixed infections with Papaya mosaic virus (PapMV), a potexvirus. These two unrelated viruses produce antagonism or synergism depending on their order of infection in the plant. When PRSV is inoculated first or at the same time as PapMV, the viral interaction is synergistic. However, an antagonistic response is observed when PapMV is inoculated before PRSV. In the antagonistic condition, PRSV is deterred from the plant and its drastic effects are overcome. Here, we examine differences in gene expression by high-throughput RNA sequencing, focused on immune system pathways. We present the transcriptomic expression of single and mixed inoculations of PRSV and PapMV leading to synergism and antagonism. Upregulation of dominant and hormone-mediated resistance transcripts suggests that the innate immune system participates in synergism. In antagonism, in addition to innate immunity, upregulation of RNA interference-mediated resistance transcripts suggests that adaptive immunity is involved.

Shifts in the diversity of an amphibian community from a premontane forest of San Ramón, Costa Rica

Biological communities are experiencing rapid shifts of composition in Neotropical ecosystems due to several factors causing population declines. However, emerging evidence has provided insights on the adaptive potential of multiple species to respond to illnesses and environmental pressures. In Costa Rica, the decline of amphibian populations is a remarkable example of these changes. Here we provide evidence of variation in the amphibian richness of a premontane forest of San Ramón (Costa Rica) across a ~30 year period. We also quantified changes in the composition and abundance of the leaf-litter frog community occurring in the same premontane forest, by comparing diversity data with a difference of ~18 years. We evaluated the similarity of species richness from 1980s to 2010s based on several sources, and the dissimilarity of species diversity in the site comparing 28 standardized surveys from 1994-1995 and 2011-2012. We compared the relative abundance of some frogs that inhabit the leaf-litter layer between these same periods. Our results show that there is more similarity in amphibian richness between 1980s and 2010s (~ 52 %) than between 1980s and 1990s (~ 40 %). The richness of leaf-litter anurans was ~ 65 % similar between 1990s and 2010s. The diversity of leaf-litter anuran was clearly different between 1994-1995 and 2011-2012, and it was clustered among those periods. We determined that the amphibian community in this premontane forest drastically changed: many species have disappeared, or gradually declined through the decades (e.g. Pristimantis ridens, P. cruentus, Craugastor bransfordii) as in other well studied localities of Costa Rica, while some few species flourished after being almost absent from the site in the 1990s (e.g. Craugastor crassidigitus, Lithobates warszewistchii). Currently dominant species such as C. crassidigitus would be using developed resistance against Bd-fungus as an advantage (apparent competition) in the premontane forest where the disease is more virulent than in lowlands. Our analysis supports the hypothesis of individualized responses of anuran populations under distinct site and elevations. We suggest to continue monitoring the amphibian communities of premontane tropical forests to understand how this ecosystem gradually resist and adapts to this catastrophic time of biodiversity loss.

Las comunidades biológicas están experimentando rápidos cambios de composición en los ecosistemas neotropicales debido a varios factores que causan disminuciones de las poblaciones. Sin embargo, la evidencia emergente ha proporcionado información sobre el potencial de adaptación de múltiples especies para responder a enfermedades y presiones ambientales. En Costa Rica, el declive de las poblaciones de anfibios es un ejemplo notable de estos cambios. Aquí proporcionamos evidencia de variación en la riqueza de anfibios de un bosque premontano de San Ramón (Costa Rica) en un período de ~ 30 años. También cuantificamos los cambios en el ensamblaje, y las oscilaciones poblacionales, de la comunidad de ranas de hojarasca que ocurren en el mismo bosque premontano, tras comparar datos de diversidad con una diferencia de ~ 18 años. Evaluamos la similitud de la riqueza de especies desde la década de 1980 hasta la de 2010 basados en varias fuentes, así como la disimilitud de la diversidad de especies en el sitio comparando 28 muestreos estandarizados entre 1994-1995 y 2011-2012. Comparamos la abundancia relativa de algunas ranas que habitan la hojarasca entre 1994-1995 y 2011-2012. Nuestros resultados muestran que existe una similitud un poco mayor en la riqueza de anfibios entre los años 1980 y 2010 que entre los 1980 y 1990 (~ 40 %). La riqueza de los anuros de la hojarasca fue aproximadamente un 65 % similar entre la década de 1990 y 2010. La diversidad de anuros de la hojarasca fue claramente diferente entre 1994-1995 y 2011-2012, y se agrupó entre esos períodos. Determinamos que la comunidad de anfibios en este bosque premontano cambió drásticamente: muchas especies han desaparecido o han disminuido gradualmente a lo largo de las décadas como en otras localidades bien estudiadas de Costa Rica (por ejemplo, Pristimantis ridens, P. cruentus, Craugastor bransfordii), mientras algunas pocas especies florecieron tras casi desaparecer de nuestro sitio en la década de 1990 (por ejemplo, Craugastor crassidigitus, Lithobates warszewistchii). Actualmente, las especies dominantes como C. crassidigitus podrían tener resistencia contra el hongo quitridio, desarrollada como una ventaja (competición aparente) en el bosque premontano donde la enfermedad es más virulenta que en las tierras bajas. Nuestro análisis apoya la hipótesis de las respuestas individualizadas de las poblaciones de anuros en distintos sitios y elevaciones. Sugerimos continuar con el monitoreo de las comunidades de anfibios de los bosques neotropicales premontanos para comprender cómo este ecosistema resiste gradualmente y se adapta a este momento catastrófico de pérdida de biodiversidad.

Morada Nova sheep breed: Resistant or resilient to Haemonchus contortus infection?

Morada Nova is a Brazilian hair sheep breed that is well adapted to the country's mainly tropical climate and has good potential for meat and leather production. This breed is reported to be resistant to Haemonchus contortus infection, a highly desired characteristic due to the large impact of this parasite on sheep farming. Therefore, the present study aimed to characterize 287 recently weaned Morada Nova lambs and 123 ewes in relation to their resistance against H. contortus. The animals were dewormed and 15 days later artificially infected with 4000 H. contortus L3 (D0). They were individually monitored by periodic assessment of fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). On D42, the sheep were again dewormed and submitted to a new parasitic challenge, following the same scheme. The animals of each category (lambs and ewes) were ranked according to individual mean FEC values, and classified as resistant (R, 20%), intermediate (I, 60%), or susceptible (S, 20%) to H. contortus infection. At weaning, high FEC were observed in all three phenotypes (P > 0.05). After the artificial infections, there was a significant difference (P < 0.05) among the three lamb phenotypes for the mean FEC (R < I < S), PCV (R > I > S), and LW (R = I > S). The infection levels (FEC) were negatively correlated with PCV (r = -0.66; P < 0.001), and LW (r = -0.30; P < 0.001). Despite this, the lambs were resilient, since more than 88% of these animals maintained the PCV above 24%, even when heavily infected. The importance of selective parasite control before weaning to reduce the negative impact on slaughter weight was evidenced, taking into account the high positive correlation between LW at weaning and final LW (r = 0.73; P < 0.001). The ewes, in turn, were strongly resistant to the parasite. Despite highly significant differences (P < 0.001) for mean FEC between phenotypes (R < I < S), 98% of the ewes maintained FEC below 4000 EPG. Their health was not affected, since PCV and LW did not differ between phenotypes, and these parameters were not significantly correlated with FEC (P > 0.05). With the phenotypic characterization performed here, it is possible to introduce procedures for parasite control in Morada Nova flocks, facilitating the target-selective treatment approach. The results of this study can also support improvement of meat production by the Morada Nova breed.

Morada Nova sheep breed: Resistant or resilient to Haemonchus contortus infection?

Morada Nova is a Brazilian hair sheep breed that is well adapted to the country's mainly tropical climate and has good potential for meat and leather production. This breed is reported to be resistant to Haemonchus contortus infection, a highly desired characteristic due to the large impact of this parasite on sheep farming. Therefore, the present study aimed to characterize 287 recently weaned Morada Nova lambs and 123 ewes in relation to their resistance against H. contortus. The animals were dewormed and 15 days later artificially infected with 4000 H. contortus L3 (D0). They were individually monitored by periodic assessment of fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). On D42, the sheep were again dewormed and submitted to a new parasitic challenge, following the same scheme. The animals of each category (lambs and ewes) were ranked according to individual mean FEC values, and classified as resistant (R, 20%), intermediate (I, 60%), or susceptible (S, 20%) to H. contortus infection. At weaning, high FEC were observed in all three phenotypes (P>0.05). After the artificial infections, there was a significant difference (P<0.05) among the three lamb phenotypes for the mean FEC (RI > S), and LW (R=I > S). The infection levels (FEC) were negatively correlated with PCV (r = -0.66; P<0.001), and LW (r = -0.30; P<0.001). Despite this, the lambs were resilient, since more than 88% of these animals maintained the PCV above 24%, even when heavily infected. The importance of selective parasite control before weaning to reduce the negative impact on slaughter weight was evidenced, taking into account the high positive correlation between LW at weaning and final LW (r=0.73; P<0.001). The ewes, in turn, were strongly resistant to the parasite. Despite highly significant differences (P<0.001) for mean FEC between phenotypes (R0.05). With the phenotypic characterization performed here, it is possible to introduce procedures for parasite control in Morada Nova flocks, facilitating the target-selective treatment approach. The results of this study can also support improvement of meat production by the Morada Nova breed.

Identification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus.

Due to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results.

Effects of protein supplementation on resistance and resilience of lambs naturally infected with gastrointestinal parasites / Efeitos da suplementação de proteínas na resistência e resiliência de cordeiros naturalmente infectados com parasitas gastrointestinais

In this study, the objective was to evaluate the efficacy of protein supplementation on gastrointestinal nematode control in lambs naturally infected. Sixty weaned Ile de France and Texel crossbred lambs, with 60 days of age, were divided into three groups (n=20) according to the level of protein in the concentrate: low protein (LP; 8.5%), moderate protein (MP; 15%), and high protein (HP; 25%). The daily amount of concentrate offered was based on 3% of live weight, which was continued throughout the experiment of 98 days. Evaluations were conducted biweekly from days 0 (D0) to 98 (D98). There were no significant differences (p > 0.05) in the average weight among the LP, MP and HP groups, which was 22.0, 21.4, and 21.1 kg, respectively, on D0 and 46.4, 48.3, and 48.2 kg, respectively, on D98. The daily weight gain was not significantly different among the groups (LP, 250 g; MP and HP, 276 g; p > 0.05). The average egg count per gram of feces (epg) of the LP group on D98 (6,765; an increase of 5,690 epg from D0) was higher than that of the MP and HP groups (1,617.5, p < 0.05 and 3,435, p > 0.05, respectively), which remained constant throughout the study. The hematocrit values decreased (p < 0.05) from D0 to D98 only in the LP group. Only in the HP group (p < 0.05), the mean serum total protein concentration increased from D0 (5.3 g dL-1) to D98 (5.8 g dL-1)...(AU)

O objetivo deste estudo foi avaliar a eficácia da suplementação de proteínas no controle de nematoides gastrointestinais em cordeiros naturalmente infectados. Foram utilizados 60 cordeiros desmamados, com 60 dias de idade, mestiços Ile de France e Texel, divididos em três grupos (n=20) de acordo com o nível de proteína no concentrado: proteína baixa (PB; 8,5%), proteína moderada (PM; 15%) e alta proteína (AP; 25%). A quantidade diária de concentrado oferecida foi baseada em 3% do peso vivo, que se manteve ao longo do período experimental de 98 dias. As avaliações foram realizadas quinzenalmente a partir do dia 0 (D0) ao dia 98 (D98). Não houve diferença (p > 0,05) no peso médio dos grupos BP, MP e AP, iniciando em D0 com 22,0 kg, 21,4 kg e 21,1 kg, e finalizando em D98 com 46,4 kg, 48,3 kg, 48,2 kg, respectivamente. Não houve diferença significativa (p > 0,05) no ganho médio de peso diário entre os diferentes grupos (PB, 250 g; PM e AP, 276 g). A contagem média de ovos por grama de fezes (opg) do grupo PB no D98 (6.665 opg, aumento de 5.690 opg em relação ao D0) foi maior que a dos grupos PM e AP (1.617,5, p < 0,05 e 3.435, p > 0,05, respectivamente), que permaneceu constante ao longo do estudo. Os valores de hematócrito diminuíram (p < 0,05), do D0 para D98, apenas no grupo PB. Somente no grupo AP (p < 0,05), a concentração sérica de proteína total aumentou...(AU)

Effects of protein supplementation on resistance and resilience of lambs naturally infected with gastrointestinal parasites / Efeitos da suplementação de proteínas na resistência e resiliência de cordeiros naturalmente infectados com parasitas gastrointestinais

In this study, the objective was to evaluate the efficacy of protein supplementation on gastrointestinal nematode control in lambs naturally infected. Sixty weaned Ile de France and Texel crossbred lambs, with 60 days of age, were divided into three groups (n=20) according to the level of protein in the concentrate: low protein (LP; 8.5%), moderate protein (MP; 15%), and high protein (HP; 25%). The daily amount of concentrate offered was based on 3% of live weight, which was continued throughout the experiment of 98 days. Evaluations were conducted biweekly from days 0 (D0) to 98 (D98). There were no significant differences (p > 0.05) in the average weight among the LP, MP and HP groups, which was 22.0, 21.4, and 21.1 kg, respectively, on D0 and 46.4, 48.3, and 48.2 kg, respectively, on D98. The daily weight gain was not significantly different among the groups (LP, 250 g; MP and HP, 276 g; p > 0.05). The average egg count per gram of feces (epg) of the LP group on D98 (6,765; an increase of 5,690 epg from D0) was higher than that of the MP and HP groups (1,617.5, p 0.05, respectively), which remained constant throughout the study. The hematocrit values decreased (p < 0.05) from D0 to D98 only in the LP group. Only in the HP group (p < 0.05), the mean serum total protein concentration increased from D0 (5.3 g dL-1) to D98 (5.8 g dL-1)...

O objetivo deste estudo foi avaliar a eficácia da suplementação de proteínas no controle de nematoides gastrointestinais em cordeiros naturalmente infectados. Foram utilizados 60 cordeiros desmamados, com 60 dias de idade, mestiços Ile de France e Texel, divididos em três grupos (n=20) de acordo com o nível de proteína no concentrado: proteína baixa (PB; 8,5%), proteína moderada (PM; 15%) e alta proteína (AP; 25%). A quantidade diária de concentrado oferecida foi baseada em 3% do peso vivo, que se manteve ao longo do período experimental de 98 dias. As avaliações foram realizadas quinzenalmente a partir do dia 0 (D0) ao dia 98 (D98). Não houve diferença (p > 0,05) no peso médio dos grupos BP, MP e AP, iniciando em D0 com 22,0 kg, 21,4 kg e 21,1 kg, e finalizando em D98 com 46,4 kg, 48,3 kg, 48,2 kg, respectivamente. Não houve diferença significativa (p > 0,05) no ganho médio de peso diário entre os diferentes grupos (PB, 250 g; PM e AP, 276 g). A contagem média de ovos por grama de fezes (opg) do grupo PB no D98 (6.665 opg, aumento de 5.690 opg em relação ao D0) foi maior que a dos grupos PM e AP (1.617,5, p 0,05, respectivamente), que permaneceu constante ao longo do estudo. Os valores de hematócrito diminuíram (p < 0,05), do D0 para D98, apenas no grupo PB. Somente no grupo AP (p < 0,05), a concentração sérica de proteína total aumentou...

Non-host Plant Resistance against Phytophthora capsici Is Mediated in Part by Members of the I2 R Gene Family in Nicotiana spp.

The identification of host genes associated with resistance to Phytophthora capsici is crucial to developing strategies of control against this oomycete pathogen. Since there are few sources of resistance to P. capsici in crop plants, non-host plants represent a promising source of resistance genes as well as excellent models to study P. capsici - plant interactions. We have previously shown that non-host resistance to P. capsici in Nicotiana spp. is mediated by the recognition of a specific P. capsici effector protein, PcAvr3a1 in a manner that suggests the involvement of a cognate disease resistance (R) genes. Here, we have used virus-induced gene silencing (VIGS) and transgenic tobacco plants expressing dsRNA in Nicotiana spp. to identify candidate R genes that mediate non-host resistance to P. capsici. Silencing of members of the I2 multigene family in the partially resistant plant N. edwardsonii and in the resistant N. tabacum resulted in compromised resistance to P. capsici. VIGS of two other components required for R gene-mediated resistance, EDS1 and SGT1, also enhanced susceptibility to P. capsici in N. edwardsonii, as well as in the susceptible plants N. benthamiana and N. clevelandii. The silencing of I2 family members in N. tabacum also compromised the recognition of PcAvr3a1. These results indicate that in this case, non-host resistance is mediated by the same components normally associated with race-specific resistance.