The role of genetic variability of the host on the resistance to Neospora caninum infection in cattle.

Neospora caninum is one of the most frequently diagnosed abortifacient pathogens in cattle. There is abundant genomic information about the parasite itself, but very little is known about the genetic variability of resistance in the most common intermediate host. The aim of this review was to compile all the available information about the genetic variability associated with the resistance to N. caninum both between and within cattle breeds. We systematically searched for published studies that investigated the influence of genetics of the host on the prevalence of N. caninum and risk of abortion. Beyond the potential confounding effects of feeding systems, management and animal density, some lines of evidence suggest that Holstein, the most popular breed for milk production, has a comparatively higher risk of abortion due to infections by N. caninum, whereas some beef breeds from Continental Europe seem to be more resistant. It is still not clear if different genetic mechanisms of resistance are involved in the two known routes of infection: postnatal ingestion of oocysts or transplacental transmission from the infected dam to the fetus. Genomic information associated with susceptibility to infection and risk of abortion in different cattle breeds is still scarce. The information reported here could be useful to identify new research alternatives and to define novel strategies to deal with this major problem of animal production.

Plant virology in the 21st century in China: Recent advances and future directions.

Plant viruses are a group of intracellular pathogens that persistently threaten global food security. Significant advances in plant virology have been achieved by Chinese scientists over the last 20 years, including basic research and technologies for preventing and controlling plant viral diseases. Here, we review these milestones and advances, including the identification of new crop-infecting viruses, dissection of pathogenic mechanisms of multiple viruses, examination of multilayered interactions among viruses, their host plants, and virus-transmitting arthropod vectors, and in-depth interrogation of plant-encoded resistance and susceptibility determinants. Notably, various plant virus-based vectors have also been successfully developed for gene function studies and target gene expression in plants. We also recommend future plant virology studies in China.

Breeding for disease resistance is an effective way to solve PRRSV.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is one of the major diseases restricting the development of large-scale pig breeding industry, which has brought huge economic losses to pig industry. Although a lot of work has been done in vaccine development, biosafety and pig health, PRRSV is characterized by easy mutation and recombination of genome, immunosuppression, enhanced antibody dependence, persistent infection, etc., making the prevention, control and elimination of PRRSV very difficult. With the deepening of PRRSV research, it is gradually realized that screening and identifying PRRSV susceptibility/resistance genes and implementing PRRSV disease resistance breeding are long-term and effective strategies for fundamental prevention and control, which has important practical significance for the prevention and control of pig herds.

Ovine PAPPA2 gene coding variants are linked to decreased fecal egg shedding in native Turkish sheep naturally infected with gastrointestinal nematodes.

In this study, the association between PAPPA2 coding variants and gastrointestinal (GI) nematode fecal egg count (FEC) score in adult Turkish sheep was investigated. For this purpose, the FEC score was determined in adult sheep from six breeds: Karacabey Merino (n = 137), Kivircik (n = 116), Cine capari (n = 109), Karakacan (n = 102), Imroz (n = 73), and Chios (n = 50). Sheep were classified as shedders or non-shedders within breeds and flocks. The first group was the fecal egg shedders (> 50 per gram of feces), and the second group was the no fecal egg shedders (≤ 50 per gram of feces). The exon 1, exon 2, exon 5, exon 7, and a part of 5'UTR of the ovine PAPPA2 gene were genotyped by Sanger sequencing of these two groups. Fourteen synonymous and three non-synonymous single-nucleotide polymorphisms (SNPs) were found. The non-synonymous SNPs, D109N, D391H, and L409R variants, are reported for the first time. Two haplotype blocks were constructed on exon 2 and exon 7. The specific haplotype, C391G424G449T473C515A542 on the exon 2 that carries the 391H variant, was tested against four other common haplotypes. Our results indicate that C391G424G449T473C515A542 haplotype was significantly associated with fecal egg shedding status in adult Turkish sheep (p-value, 0.044).

Association between ovine Toll-like receptor 4 (TLR4) gene coding variants and presence of Eimeria spp. in naturally infected adult Turkish native sheep.

Coccidiosis caused by Eimeria spp. is a protozoan disease prevalent in farm animals, and it is responsible for serious economic losses especially in young animals. It has been popular to breed disease-resistant animals due to the concern about food safety, animal welfare, and public health. Toll-like receptor (TLR) gene family plays a key role in the innate immune system participating in host-antigen interaction, therefore, they are candidate genes for breeding disease-resistant animals. In the present study, possible genetic associations between TLR4 gene coding variants and the presence of Eimeria spp. in adult Turkish sheep were investigated. For this purpose, the presence of Eimeria spp. in fecal samples from six native Turkish sheep were determined, and approximately 1450 bp region in the 3rd exon of the ovine TLR4 gene was sequenced. Ten nonsynonymous and four synonymous single nucleotide polymorphisms (SNPs) were detected in the targeted region. Statistical analyses revealed that the SNP at the codon at 356th position encoding Leucine instead of Phenylalanine (F356L) was significantly associated with the presence of Eimeria spp. It was found that the individuals carrying at least one Leucine amino acid sequence at this position have 2.3-fold more risk for the presence of Eimeria spp.

The Novel Avian Leukosis Virus Subgroup K Shares Its Cellular Receptor with Subgroup A.

Avian leukosis virus subgroup K (ALV-K) is composed of newly emerging isolates, which, in sequence analyses, cluster separately from the well-characterized subgroups A, B, C, D, E, and J. However, it remains unclear whether ALV-K represents an independent ALV subgroup with regard to receptor usage, host range, and superinfection interference. In the present study, we examined the host range of the Chinese infectious isolate JS11C1, an ALV-K prototype, and we found substantial overlap of species that were either resistant or susceptible to ALV-A and JS11C1. Ectopic expression of the chicken tva gene in mammalian cells conferred susceptibility to JS11C1, while genetic ablation of the tva gene rendered chicken DF-1 cells resistant to infection by JS11C1. Thus, tva expression is both sufficient and necessary for JS11C1 entry. Receptor sharing was also manifested in superinfection interference, with preinfection of cells with ALV-A, but not ALV-B or ALV-J, blocking subsequent JS11C1 infection. Finally, direct binding of JS11C1 and Tva was demonstrated by preincubation of the virus with soluble Tva, which substantially decreased viral infectivity in susceptible chicken cells. Collectively, these findings indicate that JS11C1 represents a new and bona fide ALV subgroup that utilizes Tva for cell entry and binds to a site other than that for ALV-A.IMPORTANCE ALV consists of several subgroups that are particularly characterized by their receptor usage, which subsequently dictates the host range and tropism of the virus. A few newly emerging and highly pathogenic Chinese ALV strains have recently been suggested to be an independent subgroup, ALV-K, based solely on their genomic sequences. Here, we performed a series of experiments with the ALV-K strain JS11C1, which showed its dependence on the Tva cell surface receptor. Due to the sharing of this receptor with ALV-A, both subgroups were able to interfere with superinfection. Because ALV-K could become an important pathogen and a significant threat to the poultry industry in Asia, the identification of a specific receptor could help in the breeding of resistant chicken lines with receptor variants with decreased susceptibility to the virus.

OXI1 kinase plays a key role in resistance of Arabidopsis towards aphids (Myzus persicae).

Plants have co-evolved with a diverse array of pathogens and insect herbivores and so have evolved an extensive repertoire of constitutive and induced defence mechanisms activated through complex signalling pathways. OXI1 kinase is required for activation of mitogen-activated protein kinases (MAPKs) and is an essential part of the signal transduction pathway linking oxidative burst signals to diverse downstream responses. Furthermore, changes in the levels of OXI1 appear to be crucial for appropriate signalling. Callose deposition also plays a key role in defence. Here we demonstrate, for the first time, that OXI1 plays an important role in defence against aphids. The Arabidopsis mutant, oxi1-2, showed significant resistance both in terms of population build-up (p < 0.001) and the rate of build-up (p < 0.001). Arabidopsis mutants for ß-1,3-glucanase, gns2 and gns3, showed partial aphid resistance, significantly delaying developmental rate, taking two-fold longer to reach adulthood. Whilst ß-1,3-glucanase genes GNS1, GNS2, GNS3 and GNS5 were not induced in oxi1-2 in response to aphid feeding, GNS2 was expressed to high levels in the corresponding WT (Col-0) in response to aphid feeding. Callose synthase GSL5 was up-regulated in oxi1-2 in response to aphids. The results suggest that resistance in oxi1-2 mutants is through induction of callose deposition via MAPKs resulting in ROS induction as an early response. Furthermore, the results suggest that the ß-1,3-glucanase genes, especially GNS2, play an important role in host plant susceptibility to aphids. Better understanding of signalling cascades underpinning tolerance to biotic stress will help inform future breeding programmes for enhancing crop resilience.

Combining a transcriptomic approach and a targeted metabolomics approach for deciphering the molecular bases of compatibility phenotype in the snail Biomphalaria glabrata toward Schistosoma mansoni.

Biomphalaria glabrata is a freshwater snail and the obligatory intermediate host of Schistosoma mansoni parasite, the etiologic agent of intestinal Schistosomiasis, in South America and Caribbean. Interestingly in such host-parasite interactions, compatibility varies between populations, strains or individuals. This observed compatibility polymorphism is based on a complex molecular-matching-phenotype, the molecular bases of which have been investigated in numerous studies, notably by comparing between different strains or geographical isolates or clonal selected snail lines. Herein we propose to decipher the constitutive molecular support of this interaction in selected non-clonal resistant and susceptible snail strain originating from the same natural population from Brazil and thus having the same genetic background. Thanks to a global RNAseq transcriptomic approach on whole snail, we identified a total of 328 differentially expressed genes between resistant and susceptible phenotypes among which 129 were up-regulated and 199 down-regulated. Metabolomic studies were used to corroborate the RNAseq results. The activation of immune genes and specific metabolic pathways in resistant snails might provide them with the capacity to better respond to parasite infection.

Prevalence and drug susceptibility of clinical Candida species in nasopharyngeal cancer patients in Vietnam.

In the nature, Candida species are normal inhabitants and can be observed in a wide variety of vertebrates. In humans, especially for cancer patients who fall prey to opportunistic pathogens, this group of susceptible multi-drug resistant and biofilm-forming yeasts, are among the commonest ones. In this study, Candida species in 76 oral lesion samples from Vietnamese nasopharyngeal-cancer patients were isolated, morphologically identified using CHROMagar™, germ tube formation, and chlamydospore formation tests, and molecularly confirmed by PCR-RFLP. The drug susceptibility of these isolates was then tested, and the gene ERG11 was DNA sequenced to investigate the mechanism of resistance. The results showed that Candida albicans remained the most prevalent species (63.16% of the cases), followed by Candida glabrata, Candida tropicalis, and Candida krusei. The rates of resistance of non-albicans Candida for tested drugs were 85.71%, 53.57%, and 57.14% to fluconazole, clotrimazole, and miconazole, respectively. Although the drug-resistance rate of Candida albicans was lower than that of non-albicans Candida, it was higher than expected, suggesting an emerging drug-resistance phenomenon. Furthermore, ERG11 DNA sequencing revealed different mutations (especially K128T), implying the presence of multiple resistance mechanisms. Altogether, the results indicate an alarming drug-resistance situation in Candida species in Vietnamese cancer patients and emphasize the importance of species identification and their drug susceptibility prior to treatment.

Role of genetic factors in different swine breeds exhibiting varying levels of resistance/susceptibility to PRRSV.

Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is an economically significant contagious disease. Traditional approaches based on vaccines or medicines were challenging to control PRRSV due to the diversity of viruses. Different breeds of pigs infected with PRRSV have been reported to have different immune responses. However, due to the complexity of interaction mechanism between host and PRRSV, the genetic mechanism leading to PRRSV susceptibility/resistance in various pig breeds is still unclear. Herein, the role of host genetic components in PRRSV susceptibility is systematically described, and the molecular mechanisms by which host genetic factors such as SNPs, cytokines, receptor molecules, intestinal flora, and non-coding RNAs regulate PRRSV susceptibility/resistance. Therefore, improving the resistance to disease of individual animals through disease-resistance breeding technology is of profound significance for uplifting the sustainable and healthy development of the pig industry.

Celastrol enhances tamoxifen sensitivity in the treatment of triple negative breast cancer via mitochondria mediated apoptosis pathway.

OBJECTIVE: To investigate the enhancement of chemosensitivity of tamoxifen (TAM) in triple negative breast cancer (TNBC) by celastrol (CEL) through mitochondrial mediated pathway. METHODS: Human TNBC MDA-MB-231 cells were divided to the control group (medium treatment), TAM-L group (low concentration TAM treatment, 0.5 µM TAM treatment), TAM-H group (1 µM TAM treatment), CEL-L group (low concentration CEL treatment, 1 µM CEL), CEL-H group (high concentration CEL treatment, 3 µM CEL), CEL-L+TAM group (low concentration CEL and TAM co-treatment) and CEL-H+TAM group (high concentration of CEL co-treated with TAM). The proliferation and invasion of cells in each cell group were detected by MTT assay and Transwell assay, respectively. Changes in mitochondrial membrane potential were determined by JC-1 staining. 2'-7'-dichlorofluorescein diacetate (DCFH-DA) fluorescence probe combined with flow cytometry was used to measure the levels of reactive oxygen species (ROS) in cells. The GSH/(GSSG+GSH) level in the cells was detected with glutathione (GSH)/oxidized glutathione (GSSG) enzyme-linked immunosorbent assay (ELISA) kit. The expression levels of the apoptosis-related proteins B lymphocytoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), sheared cysteine-containing aspartate protein hydrolase 3 (Cleaved Caspase-3), and cytochrome C (Cytc) in each group were measured by Western blot. The tumor model of subcutaneous transplantation of TNBC cells in nude mice was established. After administration, the volume and mass of tumors in each group were measured, and the tumor inhibition rate was calculated. RESULTS: Compared with the Control group, cell proliferation inhibition rate (24 h, 48 h), apoptosis rate, ROS level, Bax, cleaved caspase-3 and Cytc protein expression were conspicuously increased in the TAM, CEL-L, CEL-H, CEL-L+TAM and CEL-H+TAM groups (all P < 0.05), and cell migration and invasion, mitochondrial membrane potential, GSH level, and Bcl-2 protein expression were conspicuously decreased (all P < 0.05). Compared with the TAM group, the cell proliferation inhibition rate (24 h, 48 h), apoptosis rate, ROS level, Bax, cleaved caspase-3 and Cytc protein expression were increased in the CEL-H+TAM group (all P < 0.05), and cell migration rate, cell invasion number, mitochondrial membrane potential, GSH level and Bcl-2 protein expression were decreased in the CEL-H+TAM group (all P < 0.05). Compared with the CEL-L group, the cell proliferation inhibition rate (24 h, 48 h), apoptosis rate, ROS level, Bax, cleaved caspase-3 and Cytc protein expression were significantly increased in the CEL-H group (all P < 0.05), and cell migration rate, cell invasion number, mitochondrial membrane potential, GSH level and Bcl-2 protein expression were decreased in the CEL-H group (all P < 0.05). Compared with the model group, the tumor volume of TAM group, CEL-H group, CEL-L+TAM group and CEL-H+TAM group decreased (all P < 0.05). Compared with TAM group, the tumor volume in CEL-H+TAM group decreased significantly (P < 0.05). CONCLUSION: CEL can promote apoptosis and enhance the TAM sensitivity in TNBC treatment through a mitochondria-mediated pathway.

Mechanisms of plant cell wall surveillance in response to pathogens, cell wall-derived ligands and the effect of expansins to infection resistance or susceptibility.

Cell wall integrity is tightly regulated and maintained given that non-physiological modification of cell walls could render plants vulnerable to biotic and/or abiotic stresses. Expansins are plant cell wall-modifying proteins active during many developmental and physiological processes, but they can also be produced by bacteria and fungi during interaction with plant hosts. Cell wall alteration brought about by ectopic expression, overexpression, or exogenous addition of expansins from either eukaryote or prokaryote origin can in some instances provide resistance to pathogens, while in other cases plants become more susceptible to infection. In these circumstances altered cell wall mechanical properties might be directly responsible for pathogen resistance or susceptibility outcomes. Simultaneously, through membrane receptors for enzymatically released cell wall fragments or by sensing modified cell wall barrier properties, plants trigger intracellular signaling cascades inducing defense responses and reinforcement of the cell wall, contributing to various infection phenotypes, in which expansins might also be involved. Here, we review the plant immune response activated by cell wall surveillance mechanisms, cell wall fragments identified as responsible for immune responses, and expansin's roles in resistance and susceptibility of plants to pathogen attack.

Biochemical and molecular diagnosis of different tomato cultivars susceptible and resistant to Tuta absoluta (Meyrick) infestation.

Resistant plant cultivars which used in breeding programs are considered one of the modern integrated management programs to reduce the usage of synthetic insecticides and environmental contamination the present study aimed to characterize the resistant and susceptible tomato cultivars to Tuta absoluta based on biochemical and molecular levels, in Egypt. The biochemical characters of the tested tomato cultivars (tomato- 86, tomato- Alissa, tomato- Fayarouz, tomato- Omniya, tomato- 036, tomato- GS) were determined colorimetrically and characterized by using native- polyacrylamide gel electrophoresis (PAGE) and agarose gel. Our results showed that there were variations highly significant in all biochemical constituents of the resistant tomato cultivar (tomato- 86) compared with the susceptible one (tomato- GS). Also, native-(PAGE) for peroxidase (POD) isoenzymes techniques of the tested tomato cultivars showed variations in protein band numbers and densities in tomato-86 resistant compared with tomato-GS susceptible to Tuta absoluta infestation. The correlation coefficient between total phenols and peroxidases in infested tomato leaves and percentages of damaged leaves with the tested insect pest was negative and highly significant, while in case of total proteins and reducing sugars in infested tomato leaves as well as lycopene contents in infested tomato fruits was positive, highly significant and significant, respectively. The correlation coefficient between tomato yield means and the infested fruit percentage with T. absoluta larvae was negative and highly significant. Respecting molecular diagnosis random amplified polymorphism DNA- polymerase chain reaction (RAPD- PCR), the results demonstrated that the presence of polymorphism in the resistant tomato cultivar (tomato- 86) compared with (tomato- GS), the most susceptible to the tested insect pest infestation.

Symptomless Resistance of Alfalfa to Meloidogyne incognita acrita.

Penetration, development and migration of the cotton root-knot nematode, Meloidogyne incognita acrita, in resistant and susceptible alfalfa varieties was compared. Larvae entered both resistant and susceptible plants in approximately the same numbers. After 3 to 4 days, the number of larvae in resistant roots decreased sharply until at 7 days fewer than 5 larvae/seedling and no nematode development could be found. In susceptible roots, larvae became sedentary and developed normally; egg production began as early as 18 days after penetration of the host.

Molecular basis for benzimidazole resistance from a novel ß-tubulin binding site model.

Benzimidazole-2-carbamate derivatives (BzCs) are the most commonly used antiparasitic drugs for the treatment of protozoan and helminthic infections. BzCs inhibit the microtubule polymerization mechanism through binding selectively to the ß-tubulin subunit in which mutations have been identified that lead to drug resistance. Currently, the lack of crystallographic structures of ß-tubulin in parasites has limited the study of the binding site and the analysis of the resistance to BzCs. Recently, our research group has proposed a model to explain the interaction between the BzCs and a binding site in the ß-tubulin. Herein, we report the homology models of two susceptible (Haemonchus contortus and Giardia intestinalis) parasites and one unsusceptible (Entamoeba histolytica) generated using the structure of the mammal Ovis aries, considered as a low susceptible organism, as a template. Additionally, the mechanism by which the principal single point mutations Phe167Tyr, Glu198Ala and Phe200Tyr could lead to resistance to BzCs is analyzed. Molecular docking and molecular dynamics studies were carried out in order to evaluate the models and the ligand-protein complexes' behaviors. This study represents a first attempt towards understanding, at the molecular level, the structural composition of ß-tubulin in all organisms, also suggesting possible resistance mechanisms. Furthermore, these results support the importance of benzimidazole derivative optimization in order to design more potent and selective (less toxic) molecules for the treatment of parasitic diseases.

Variability in the fecal egg count and the parasitic burden of hair sheep after grazing in nematode infected paddocks

This study aimed to evaluate the variability in the fecal egg count (FEC) and the parasitic burden of naive hair sheep after grazing in nematode infected paddocks. The research was carried out in Tabasco, Mexico, during two periods (August and December). In each period 32 lambs were grazed for one month on African star grass (Cynodon plectostachyus) contaminated with gastrointestinal parasitic nematodes. FEC, packed cell volume (PCV) and body weight (BW) were recorded. Gastrointestinal worms were recovered at necropsy. Data were analyzed with the MIXED procedure of SAS using a model of repeated measurements over time. A higher number of Haemonchus contortus worms was found in December (2814±838) than in August (1166±305). The opposite occurred with Cooperia curticei (2167±393 and 3638±441, respectively). The FEC and correlation coefficient in respect to the worm burden were higher in December (6516 ± 1599, r=0.83, respectively) than in August (4364±771, r=0.44, respectively). A high variability in resistance-susceptibility to gastrointestinal nematodes (GIN) occurred in Katahdin × Pelibuey lambs after grazing.