The phytopathogen Xanthomonas campestris senses and effluxes salicylic acid via a sensor HepR and an RND family efflux pump to promote virulence in host plants.

Salicylic acid (SA) plays an essential role in plant defense against biotrophic and semi-biotrophic pathogens. Following pathogen recognition, SA biosynthesis dramatically increases at the infection site of the host plant. The manner in which pathogens sense and tolerate the onslaught of SA stress to survive in the plant following infection remains to be understood. The objective of this work was to determine how the model phytopathogen Xanthomonas campestris pv. campestris (Xcc) senses and effluxes SA during infection inside host plants. First, RNA-Seq analysis identified an SA-responsive operon Xcc4167-Xcc4171, encoding a MarR family transcription factor HepR and an RND (resistance-nodulation-cell division) family efflux pump HepABCD in Xcc. Electrophoretic mobility shift assays and DNase I footprint analysis revealed that HepR negatively regulated hepABCD expression by specifically binding to an AT-rich region of the promoter of the hepRABCD operon, Phep. Second, isothermal titration calorimetry and further genetic analysis suggest that HepR is a novel SA sensor. SA binding released HepR from its cognate promoter Phep and then induced the expression of hepABCD. Third, the RND family efflux pump HepABCD was responsible for SA efflux. The hepRABCD cluster was also involved in the regulation of culture pH and quorum sensing signal diffusible signaling factor turnover. Finally, the hepRABCD cluster was transcribed during the XC1 infection of Chinese radish and was required for the full virulence of Xcc in Chinese radish and cabbage. These findings suggest that the ability of Xcc to co-opt the plant defense signal SA to activate the multidrug efflux pump may have evolved to ensure Xcc survival and virulence in susceptible host plants.

Exploiting the roles of nitrogen sources for HEA increment in Cordyceps cicadae.

Cordyceps cicadae, as a new food ingredient, is a valuable edible and medicinal fungi. However, its resources are severely depleted due to environmental limitations and excessive harvesting practices. N6-(2-hydroxyethyl) adenosine (HEA), as an important product of Cordyceps cicadae, has the potential to be used in medical industry due to its diverse disease curing potential. However, the disclosure of HEA synthesis still severely limited its application until now. In this study, the kinetic curves for adenosine and HEA under shaker fermentation were explored. The kinetics of HEA and adenosine production exhibited a competitive pattern, implicating a possibility of sharing a same step during their synthesis. Due to HEA as a derivative of nitrogen metabolism, the effect of different nitrogen sources (peptone, yeast extract, ammonium sulfate, diammonium oxalate monohydrate, ammonium citrate dibasic, and ammonium citrate tribasic) on HEA production in Cordyceps cicadae strain AH 10-4 had been explored under different incubation conditions (shaker fermentation, stationary fermentation, and submerged fermentation). Our results indicated that the complex organic nitrogen sources were found to improve the accumulation of HEA content under shaker fermentation. In contrast, the optimal nitrogen source for the accumulation of HEA under stationary fermentation and submerged fermentation was ammonium citrate tribasic. But submerged fermentation obviously shortened the incubation time and had a comparable capacity of HEA accumulation by 2.578 mg/g compared with stationary fermentation of 2.535 mg/g, implicating a possibility of scaled-up production of HEA in industry by submerged fermentation. Based on the dramatic HEA production by ammonium sulfate as nitrogen resources between stationary and shaker fermentations, alanine, aspartate and glutamate as well as arginine metabolic pathway were related to the production of HEA by comparative transcriptome. Further investigation indicated that glutamic acid, which is an analog of Asp, showed an optimum production of HEA in comparison with other amino acids.

Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.

Filamentous fungi play a crucial role in environmental pollution control, protein secretion, and the production of active secondary metabolites. The evolution of gene editing technology has significantly improved the study of filamentous fungi, which in the past was laborious and time-consuming. But recently, CRISPR-Cas systems, which utilize small guide RNA (sgRNA) to mediate clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas), have demonstrated considerable promise in research and application for filamentous fungi. The principle, function, and classification of CRISPR-Cas, along with its application strategies and research progress in filamentous fungi, will all be covered in the review. Additionally, we will go over general matters to take into account when editing a genome with the CRISPR-Cas system, including the creation of vectors, different transformation methodologies, multiple editing approaches, CRISPR-mediated transcriptional activation (CRISPRa) or interference (CRISPRi), base editors (BEs), and Prime editors (PEs).

Differences in Toxoplasma gondii distribution in different muscle and viscera of naturally infected sheep.

Toxoplasma gondii (T. gondii) is a zoonotic parasite that can cause serious pathology in intermediate hosts such as humans and animals. Eating undercooked or raw meat is the most important route of infection by T. gondii. Sheep are an important source of meat worldwide, and they are also susceptible to T. gondii. Mutton infected with T. gondii poses a serious threat to the food safety of consumers. At present, studies have mainly focused on the infection ratio of T. gondii in livestock; however, systematic studies have not been performed on differences in the distribution of this parasite in different muscle and viscera tissues of animals. In this study, the differences in the distribution of T. gondii in naturally infected Small-tailed Han sheep was studied. By amplifying the B1 gene of the parasite via real-time fluorescence quantification PCR (RT‒qPCR), we found that the parasite burden of T. gondii differed among different parts of the sheep, with the highest burden observed in the heart among the viscera and the external ridge among the muscle. The relative expression was ranked from high to low in our study as follows: heart, spleen, external ridge, tenderloin, lung, liver, kidney, neck meat, forelegs, cucumber strips, hind leg, lamb belly, and lamb chops. This study provided important guidance for monitoring the food safety of mutton products.

Study on the effect of koumiss on reactivation of Toxoplasma gondii infection.

Toxoplasma gondii is an obligate intracellular parasite that infects nucleated cells of all warm-blooded animals, and most patients have latent infections. The latent infection will be reactivated in the immunocompromised or immunocompromised individuals, which will lead to severe toxoplasmosis. At present, less research has been focused on the reactivation of T. gondii infection. Koumiss is a kind of fermented milk made from fresh mare's milk through natural fermentation that can be applied to clinical and rehabilitation medicine to mitigate the development of various diseases due to its unique functional characteristics. In this study, we explored the antagonistic effect of koumiss on reactivation of T. gondii infection. Mice were treated with dexamethasone to establish a reactivation model after infection with T. gondii and then treated with koumiss. The survival rate, SHIRPA test, serum cytokine levels, organ parasite burden and intestinal microbiota were measured, respectively. Our results showed that koumiss treatment improved the clinical symptoms of mice, significantly reduced the organ parasite burden of mice, and improved the composition and structure of intestinal flora. This study provides new evidence for the alleviation and treatment of toxoplasmosis and provides a novel idea for the development and utilization of koumiss.

Study on the antagonistic effects of koumiss on Toxoplasma gondii infection in mice.

Toxoplasma gondii is an important food-borne zoonotic parasite, and approximately one-third of people worldwide are positive for T. gondii antibodies. To date, there are no specific drugs or vaccines against T. gondii. Therefore, developing a new safe and effective method has become a new trend in treating toxoplasmosis. Koumiss is rich in probiotics and many components that can alleviate the clinical symptoms of many diseases via the functional characteristics of koumiss and its regulation of intestinal flora. To investigate the antagonistic effect of koumiss on T. gondii infection, the model of acute and chronic T. gondii infection was established in this study. The survival rate, SHIRPA score, serum cytokine levels, brain cyst counts, ß-amyloid deposition and intestinal flora changes were measured after koumiss feeding. The results showed that the clinical symptoms of mice were improved at 6 dpi and that the SHIRPA score decreased after koumiss feeding (P < 0.05). At the same time, the levels of IL-4, IFN-γ and TNF-α decreased (P < 0.001, P < 0.001, P < 0.01). There was no significant difference of survival rate between koumiss treatment and the other groups. Surprisingly, the results of chronic infection models showed that koumiss could significantly reduce the number of brain cysts in mice (P < 0.05), improve ß-amyloid deposition in the hippocampus (P < 0.01) and decrease the levels of IFN-γ and TNF-α (P < 0.01, P < 0.05). Moreover, koumiss could influence the gut microbiota function in resisting T. gondii infection. In conclusion, koumiss had a significant effect on chronic T. gondii infection in mice and could improve the relevant indicators of acute T. gondii infection in mice. The research provides new evidence for the development of safe and effective anti-T. gondii methods, as well as a theoretical basis and data support for the use of probiotics against T. gondii infection and broadened thoughts for the development and utilization of koumiss.

Effect of collagen casing on the quality characteristics of fermented sausage.

OBJECTIVE: Fermented sausage is popular all over the world for its rich nutrition and unique flavor. Sausage casing is one of the key factors affecting the quality of fermented sausage. However, there is little information involved in this field. METHODS: In this study, collagen casings were used as a wrapping material, and natural casings (pig casings) were used as a control. The effects of the two types of casings on biogenic amine content and other quality characteristics of fermented sausage were analyzed with increasing the storage time. RESULTS: The results showed that with storage time increasing, the hardness and gumminess of fermented sausage in collagen casing (CC) group were higher than those in pig casing (PC) group (P<0.05), while the elasticity in CC group was lower than that in PC group (P<0.05). In the processing and storage period, there was no significant difference in the type and content of flavor substances between the two groups. More importantly, the contents of tryptamine, putrescine, cadaverine, histamine, tyramine and phenyethylamine in fermented sausage of CC group were lower than those in PC group (P<0.05). CONCLUSION: In conclusion, this study revealed that CC could improve the quality characteristics of fermented sausage and reduce the content of biogenic amines in fermented sausage, which provides a theoretical basis for the choice of casings in industrial production in the future.

Study on the effect of koumiss on the intestinal microbiota of mice infected with Toxoplasma gondii.

Toxoplasma gondii is a worldwide food-borne parasite that can infect almost all warm-blooded animals, including humans. To date, there are no effective drugs to prevent or eradicate T. gondii infection. Recent studies have shown that probiotics could influence the relationship between the microbiota and parasites in the host. Koumiss has been used to treat many diseases based on its probiotic diversity. Therefore, we explored the effect of koumiss on T. gondii infection via its effect on the host intestinal microbiota. BALB/c mice were infected with T. gondii and treated with PBS, koumiss and mares' milk. Brain cysts were counted, and long-term changes in the microbiota and the effect of koumiss on gut microbiota were investigated with high-throughput sequencing technology. The results suggested that koumiss treatment significantly decreased the cyst counts in the brain (P < 0.05). Moreover, T. gondii infection changed the microbiota composition, and koumiss treatment increased the relative abundance of Lachnospiraceae and Akkermansia muciniphila, which were associated with preventing T. gondii infection. Moreover, koumiss could inhibit or ameliorate T. gondii infection by increasing the abundance of certain bacteria that control unique metabolic pathways. The study not only established a close interaction among the host, intracellular pathogens and intestinal microbiota but also provided a novel focus for drug development to prevent and eradicate T. gondii infection.

An Investigation of the Relationship between Cyniclomyces guttulatus and Rabbit Diarrhoea.

Cyniclomyces guttulatus is usually recognised as an inhabitant of the gastrointestinal (GI) tract in rabbits. However, large numbers of C. guttulatus are often detected in the faeces of diarrhoeic rabbits. The relationship of C. guttulatus with rabbit diarrhoea needs to be clearly identified. In this study, a C. guttulatus Zhejiang strain was isolated from a New Zealand White rabbit with severe diarrhoea and then inoculated into SPF New Zealand white rabbits alone or co-inoculated with Eimeriaintestinalis, another kind of pathogen in rabbits. Our results showed that the optimal culture medium pH and temperature for this yeast were pH 4.5 and 40-42 °C, respectively. The sequence lengths of the 18S and 26S ribosomal DNA fragments were 1559 bp and 632 bp, respectively, and showed 99.8% homology with the 18S ribosomal sequence of the NRRL Y-17561 isolate from dogs and 100% homology with the 26S ribosomal sequence of DPA-CGR1 and CGDPA-GP1 isolates from rabbits and guinea pigs, respectively. In animal experiments, the C. guttulatus Zhejiang strain was not pathogenic to healthy rabbits, even when 1 × 108 vegetative cells were used per rabbit. Surprisingly, rabbits inoculated with yeast showed a slightly better body weight gain and higher food intake. However, SPF rabbits co-inoculated with C. guttulatus and E. intestinalis developed more severe coccidiosis than rabbits inoculated with C. guttulatus or E. intestinalis alone. In addition, we surveyed the prevalence of C. guttulatus in rabbits and found that the positive rate was 83% in Zhejiang Province. In summary, the results indicated that C. guttulatus alone is not pathogenic to healthy rabbits, although might be an opportunistic pathogen when the digestive tract is damaged by other pathogens, such as coccidia.

Exogenous nitric oxide stimulates early egress of Eimeria tenella sporozoites from primary chicken kidney cells in vitro.

Egress plays a vital role in the life cycle of apicomplexan parasites including Eimeria tenella, which has been attracting attention from various research groups. Many recent studies have focused on early egress induced by immune molecules to develop a new method of apicomplexan parasite elimination. In this study, we investigated whether nitric oxide (NO), an immune molecule produced by different types of cells in response to cytokine stimulation, could induce early egress of eimerian sporozoites in vitro. Eimeria tenella sporozoites were extracted and cultured in primary chicken kidney cells. The number of sporozoites egressed from infected cells was analyzed by flow cytometry after treatment with NO released by sodium nitroferricyanide (II) dihydrate. The results showed that exogenous NO stimulated the rapid egress of E. tenella sporozoites from primary chicken kidney cells before replication of the parasite. We also found that egress was dependent on intra-parasitic calcium ion (Ca2+) levels and no damage occurred to host cells after egress. The virulence of egressed sporozoites was significantly lower than that of fresh sporozoites. The results of this study contribute to a novel field examining the interactions between apicomplexan parasites and their host cells, as well as that of the clearance of intracellular pathogens by the host immune system.

TITLE: L'oxyde nitrique exogène stimule in vitro la sortie précoce des sporozoïtes d'Eimeria tenella des cellules primaires de rein de poulet. ABSTRACT: La sortie des cellules joue un rôle vital dans le cycle de vie des parasites Apicomplexa, y compris Eimeria tenella, ce qui a attiré l'attention de plusieurs groupes de recherche. De nombreuses études récentes se sont concentrées sur la sortie précoce induite par des molécules immunitaires, pour développer une nouvelle méthode d'élimination des parasites Apicomplexa. Dans cette étude, nous avons examiné si l'oxyde nitrique (NO), une molécule immunitaire produite par différents types de cellules en réponse à la stimulation des cytokines, pouvait induire in vitro une sortie précoce des sporozoïtes des Eimeria. Les sporozoïtes d'E. tenella ont été extraits et cultivés dans des cellules primaires de rein de poulet. Le nombre de sporozoïtes sortant des cellules infectées a été analysé par cytométrie en flux après traitement avec du NO libéré par le nitroferricyanure de sodium (II) dihydraté. Les résultats ont montré que le NO exogène stimulait la sortie rapide des sporozoïtes d'E. tenella des cellules primaires de rein de poulet avant la réplication du parasite. Nous avons également constaté que la sortie dépendait des niveaux intra-parasitaires d'ions calcium (Ca2+) et qu'aucun dommage n'est survenu aux cellules hôtes après la sortie. La virulence des sporozoïtes sortis était significativement inférieure à celle des sporozoïtes frais. Les résultats de cette étude contribuent à un nouveau domaine d'étude des interactions entre les parasites Apicomplexa et leurs cellules hôtes, ainsi qu'à celui relatif à l'élimination des pathogènes intracellulaires par le système immunitaire de l'hôte.

Seroprevalence of Toxoplasma gondii infection in sheep in Inner Mongolia Province, China.

Toxoplasma gondii is an important zoonotic parasite that can infect almost all warm-blooded animals, including humans, and infection may result in many adverse effects on animal husbandry production. Animal husbandry in Inner Mongolia is well developed, but data on T. gondii infection in sheep are lacking. In this study, we determined the seroprevalence and risk factors associated with the seroprevalence of T. gondii using an indirect enzyme-linked immunosorbent assay (ELISA) test. A total of 1853 serum samples were collected from 29 counties of Xilin Gol League (n = 624), Hohhot City (n = 225), Ordos City (n = 158), Wulanchabu City (n = 144), Bayan Nur City (n = 114) and Hulunbeir City (n = 588). The overall seroprevalence of T. gondii was 15.43%. Risk factor analysis showed that seroprevalence was higher in sheep ≥12 months of age (21.85%) than that in sheep <12 months of age (10.20%) (p < 0.01). Seroprevalence was higher in male sheep (18.76%) than females (12.80%) (p < 0.01). Barn-feeding sheep (23.13%) had higher prevalence than grazing sheep (10.94%) (p < 0.01). The seroprevalence was significantly different in different districts (p < 0.01). This study shows that sheep are exposed to T. gondii in Inner Mongolia, and provides a data reference for public health and disease control.

TITLE: Séroprévalence de l'infection par Toxoplasma gondii des moutons dans la province de Mongolie intérieure, Chine. ABSTRACT: Toxoplasma gondii est un parasite zoonotique important qui peut infecter presque tous les animaux à sang chaud, y compris les humains, et son infection peut entraîner de nombreux effets néfastes sur la production animale. L'élevage de Mongolie intérieure est développé, mais les données sur l'infection des moutons par T. gondii manquent. Dans cette étude, nous avons déterminé la séroprévalence et les facteurs de risque associés à la séroprévalence de T. gondii en utilisant un test immuno-enzymatique indirect (ELISA). Un total de 1853 échantillons de sérum ont été prélevés dans vingt-neuf comtés de la Ligue Xilin Gol (n = 624), Hohhot City (n = 225), Ordos City (n = 158), Wulanchabu City (n = 144), Bayan Nur City (n = 114) et la ville de Hulunbeir (n = 588). La séroprévalence globale de T. gondii était de 15,43 %. L'analyse des facteurs de risque a montré que le taux d'infection était plus élevé chez les ovins ≥ 12 mois (21,85 %) que chez les ovins <12 mois (10,20 %) (p < 0,01). Les moutons mâles (18,76 %) avaient une séroprévalence plus élevée que les femelles (12,80 %) (p < 0,01). Les moutons nourris à l'étable (23,13 %) avaient une séroprévalence plus élevée que ceux au pâturage (10,94 %) (p < 0,01). La séroprévalence était significativement différente entre les différents districts (p < 0,01). L'étude a montré que les moutons étaient exposés à T. gondii en Mongolie intérieure et a fourni une référence de données pour la santé publique et la lutte contre les maladies.

A Kinect-based upper limb rehabilitation system to assist people with cerebral palsy.

This study assessed the possibility of rehabilitating two adolescents with cerebral palsy (CP) using a Kinect-based system in a public school setting. The system provided 3 degrees of freedom for prescribing a rehabilitation program to achieve customized treatment. This study was carried out according to an ABAB reversal replication design in which A represented the baseline and B represented intervention phases. Data showed that the two participants significantly increased their motivation for upper limb rehabilitation, thus improving exercise performance during the intervention phases. Practical and developmental implications of the findings are discussed.