Astrocyte Control of Zika Infection Is Independent of Interferon Type I and Type III Expression

Zika virus (ZIKV) is a pathogenic neurotropic virus that infects the central nervous system (CNS) and results in various neurological complications. Astrocytes are the dominant CNS cell producer of the antiviral cytokine IFN-β, however little is known about the factors involved in their ability to mediate viral infection control. Recent studies have displayed differential responses in astrocytes to ZIKV infection, and this study sought to elucidate astrocyte cell-specific responses to ZIKV using a variety of cell models infected with either the African (MR766) or Asian (PRVABC59) ZIKV strains. Expression levels of pro-inflammatory (TNF-α and IL-1β) and inflammatory (IL-8) cytokines following viral infection were low and mostly comparable within the ZIKV-resistant and ZIKV-susceptible astrocyte models, with better control of proinflammatory cytokines displayed in resistant astrocyte cells, synchronising with the viral infection level at specific timepoints. Astrocyte cell lines displaying ZIKV-resistance also demonstrated early upregulation of multiple antiviral genes compared with susceptible astrocytes. Interestingly, pre-stimulation of ZIKV-susceptible astrocytes with either poly(I:C) or poly(dA:dT) showed efficient protection against ZIKV compared with pre-stimulation with either recombinant IFN-β or IFN-λ, perhaps indicating that a more diverse antiviral gene expression is necessary for astrocyte control of ZIKV, and this is driven in part through interferon-independent mechanisms.

Systemic infection of SARS-CoV-2 in free ranging Leopard (Panthera pardus fusca) in India

We report patho-morphological and virological characterization of SARS-CoV-2 in naturally infected, free ranging Indian Leopard (Panthera pardus fusca). Whole genome sequence analysis confirmed infection of Delta variant of SARS-CoV-2, possibly spill over from humans, but the case was detected when infection level had dropped significantly in human population. This report underlines the need for intensive screening of wild animals for keeping track of the virus evolution and development of carrier status of SARS-CoV-2 among wildlife species.

Lactiplantibacillus plantarum 1201 Inhibits Intestinal Infection of Salmonella enterica subsp. Enterica serovar Typhimurium Strain ATCC 13311 in Mice with High-Fat Diet

Salmonella Typhimurium is widely distributed in food. It can colonise the gastrointestinal tract after ingestion, causing lamina propria edema, inflammatory cell infiltration, and mucosal epithelial decomposition. A high-fat diet (HFD) can induce an inflammatory response, but whether HFD can increase the infection level of S. Typhimurium is unknown. We established a model of Salmonella enterica subsp. enterica serovar Typhimurium strain ATCC 13311 ATCC 13311 infection in healthy adult mice with a maintenance diet (MD) or HFD to explore the effect of Lactiplantibacillus plantarum 1201 intervention on S. Typhimurium ATCC 13311 colonization and its protective effects on mice. HFD exacerbated the infection of S. Typhimurium ATCC 13311, while the intervention of L. plantarum 1201 effectively mitigated this process. L. plantarum 1201 can reduce the colonies of S. ATCC 13311 in the intestines and tissues; and reduce intestinal inflammation by down-regulating the level of TLR4/NF-κB pathway related proteins in serum and the expression of related inflammatory factors in the colon and jejunum. Since L. plantarum 1201 can inhibit the colonization of S. Typhimurium ATCC 13311 and relieve inflammation in HFD, current research may support the use of L. plantarum 1201 to prevent S. Typhimurium infection.

Preliminary study of homeopathic treatment of subclinical mastitis evaluated through somatic cells count (SCC) and California mastitis test (CMT)

The present paper presents the results of homeopathic treatment of 25 Holstein breed cows aged 3 to 8 years old diagnosed with subclinical mastitis through California Mastitis Test (CMT). Animals were divided into 3 groups according with infection level. A homeopathic complex was developed on the grounds of clinical aspects, including Phosphorus 30x, Phytolacca 30x, Silicea 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Pulsatilla 30x, Calendula 30x and biotherapic of Staphylococcus aureus 200x. The remedy was added to salt and was administered to cattle 100g/cow/day for 75 days. CMT were carried out every 2 weeks to control incidence and severity of mastitis; somatic cells count (SCC) was performed at the beginning and the end of treatment. CMT showed significant improvement in regression of infection level all throughout the study; final SCC showed decrease in 82% of animals, signaling thus efficacy of the homeopathic treatment. Keywords: Subclinical mastitis; Dairy cows; Homeopathy; California Mastitis Test, Somatic Cells Count.  Estudo preliminar do tratamento homeopático da mastite subclínica avaliado por contagem de células somáticas (SCC) e California Mastitis Test (CMT) Resumo Este artigo apresenta os resultados do tratamento homeopático de 25 vacas raça Holstein entre 3 e 8 anos de idade diagnosticadas com mastite subclínica através do California Mastitis Test (CMT). Os animais foram divididos em 3 grupos de acordo com o nível da infecção. Foi preparado um complexo homeopático com base nos achados clínicos, composto de: Phosphorus 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x s 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x e bioterápico de Staphylococcus aureus 200x. O complexo foi acrescentado no sal e administrado na dose de 100g/vaca/dia. O CMT foi realizado a cada 2 semanas a fim de monitorar a incidéncia e gravidade da mastitie; a contagem de células somáticas (SCC) foi realizada ao início e no final do tratamento. Os valores do CMT mostraram melhora significativa no sentido de regressão do nível da infecção ao longo do estudo; o valor final da SCC diminuiu em 82% dos animais, apontando para a eficácia do tratamento homeopático. Palavras-chave: Mastite subclínica; Vacas leiteiras; Homeopatia; CMT, contagem de células somáticas.  Estudio preliminar del tratamiento homeopático de la mastitis sublínica evaluado mediante recuento de células somáticas (SCC) y California Mastitis Test (CMT) Resumen Este artículo presenta los resultados del tratamiento homeopático de 25 vacas raza Holstein entre 3 e 8 años de edad diagnosticadas com mastitis sucblínica mediante California Mastitis Test (CMT). Los animales fueron divididos en 3 grupos según el nível de la infección. Fue compuesto un complejo homeopático con base en la presentación clínica, incluyendo Phosphorus 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x y bioterápico de Staphylococcus aureus 200x. El complejo fue agregado en sal y administrado a los animales en la dosis de 100g/vaca/día. CMT fue realizado cada 2 semanas para monitorizar la incidencia y gravedad de la mastitis; recuento de células somáticas (SCC) fue realizado al inicio y al final del tratamiento. CMT mostró mejoría significativa en el sentido de regresión del nivel de la infección a lo largo del estudio; el valor final del SCC disminuyó en 82% de los animales, apuntando para la eficacia del tratamiento homeopático. Palabras-clave: Mastitis subclínica; Vacas lecheras; Homeopatía; CMT, recuento de células somáticas.  Correspondence author: Zilda Cristiani Gazim, [email protected] How to cite this article: Barzon CD, Medeiros F, Moraes RE, Silva LCM, Massambani C, Takemura OS, Gazim ZC. Preliminary study of homeopathic treatment of subclinical mastitis evaluated through somatic cells count and California mastitis test. Int J High Dilution Res [online]. 2008 [cited YYYY Mmm DD]; 7(24): 147-151. Available from: http://journal.giri-society.org/index.php/ijhdr/article/view/292/364. ÂÂÂ

Phytopathological and nutraceutical evaluation of cauliflower plants treated with high dilutions of arsenic trioxide

Introduction: This research aimed at verifying the effects of highly diluted (HD) treatments on cauliflower (Brassica oleracea L.) plants both healthy and inoculated by the fungus Alternaria brassicicola, causing the dark leaf spot disease. In vitro spore germination assays (A), growth chamber experiments (B) and field trials (C) were performed. Material and Methods: (A): spore suspensions were prepared in HD treatments and their inhibiting effect on germination was recorded microscopically after incubation at 25°C for 5 h. (B): the same treatments were tested in plants artificially inoculated with the fungus. The infection level on leaves was blindly evaluated by a previously defined infection scale. (C): the field was divided into plots according to a complete randomized block design. In the first trial (i), plants were artificially inoculated and weekly treated; the infection level was evaluated on cauliflower heads. The second trial (ii) was performed on the same field with the aim to induce a natural infection, mediated by infected crop residues. Measurement endpoints concerned the evaluation of some physiological parameters along with the glucosinolate content on cauliflower heads. Results: (A): arsenic trioxide (As 35x and 35x diluted 1:5000) and Cuprum 5x induced highly significant inhibition of germination rate (-60%) vs. control. (B): As 35x and Cu 3 g/l induced a significant decrease of mean infection level (-50%). (C): in (i), a significant reduction of disease symptoms on heads was recorded for As 35x and Cu 3 g/l (-45%). In (ii) natural fungal infection did not occur due to dry weather conditions; physiological and nutraceutical analyses of healthy heads demonstrated that As 35x induced a significant increase of both head size and glucosinolate content. Discussion: Some evidences on the efficacy of arsenic, at different decimal and centesimal HD, in fungal and viral disease control were previously reported [1]. In the present study the efficacy of HD arsenic in dark leaf spot control in field has been shown for the first time: since fungal inoculation was performed on the leaves before flowering, we can hypothesize that this treatment induced an increase of plant resistance to fungal infection. Conclusions: This research showed the possibility of using HD arsenic in agriculture (“agrohomeopathy”), as it increased both plant resistance to fungal infection and the content of glucosinolates, ie secondary metabolites involved in plant resistance mechanisms [2] and considered as “plant food protection agents” [3]. Acknowledgments: Authors declare there is no conflict of interest. This research has been supported by Marche Region. A grateful acknowledgement to Dr. Leonardo Valenti for his support to this research. The authors thank Laboratoires Boiron srl for the grant awarded to one of the author Dr. Grazia Trebbi. Finally, authors are grateful to Cemon srl for financial support of glucosinolate analyses. We had full access to all the data in this study and we take responsibility for the integrity of the data and the accuracy of the data analysis Keywords: cauliflower, arsenic trioxide, Alternaria brassicicola, glucosinolates References [1] Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jäger T, et al. Use of homeopathic preparations in phytopathological models and in field trials: a critical review. Homeopathy 2009; 98: 244-266. [2] Ménard R, Larue J-P, Silué D, Thouvenot D. Glucosinolates in cauliflower as biochemical markers for resistance against downy mildew. Phytochemistry 1999; 52: 29-35. [3] Talalay P, Fahey JW. Phytochemicals from Cruciferous plants protect against cancer by modulating carcinogen metabolism. J Nutr. 2001; 131:3027S- 3033S.

165. Decreased Antimicrobial Consumption and Decreased Rates of Multi-drug Resistant Organisms Following Onset of the COVID-19 Pandemic: Experience from an Australian Tertiary Hospital

Background Current guidelines recommend empiric antibiotics be used only for severe cases of coronavirus disease 2019 (COVID-19) or in cases where there is high clinical suspicion for bacterial co-infection. Level of adherence to guideline-recommended prescribing is unknown and high rates of antimicrobial prescribing may lead to increased development of resistance. Methods We reviewed antimicrobial prescribing patterns for patients with COVID-19 managed at The Alfred Hospital in Melbourne, Australia in 2020. Adherence to World Health Organization (WHO) guideline-based prescribing was assessed by manual review of case notes. Monthly hospital-wide antibacterial consumption April-Dec 2020 (post-pandemic period) was compared to Jan 2019-Mar 2020 (pre-pandemic period), measured as days of therapy (DOT) per 1000 patient-days. Rates of multi-drug resistant organisms (MRO) (including MRSA, VRE, CPE, ESBL) were compared between months in 2019 and 2020 after pandemic onset (April 2020) and expressed as isolates per 1000 patient-days. Results 147 patients were managed for COVID-19 in 2020 at our centre. 101 patients required hospital admission and 58 (39%) were classified as either severe or critical in severity. 80 (54%) patients received empiric antimicrobial treatment, including 78/101 (77%) of hospital inpatients and 24/26 (92%) of ICU-admitted patients. 59 (73%) of antimicrobial prescriptions were adherent to WHO guidelines. Monthly antibacterial consumption was significantly lower post-pandemic than in the pre-pandemic period (mean 853 vs 902 DOT/1000 patient-days, P=0.0065). Antimicrobial use patterns varied, with significant decreases in commonly used antibiotics such as ceftriaxone, piperacillin-tazobactam, azithromycin and ciprofloxacin but no change in vancomycin or meropenem (Figure 1). There was a mean decrease of 0.77 MRO isolates/1000 patient-days (P=0.026) when each month in 2020 was compared with the corresponding month in 2019 (Figure 2). Antibacterial consumption in 2019 and 2020 by month, expressed as days of therapy/1000 patient-days. Rates of isolated multi-drug resistant organisms in 2019 and 2020 by month, expressed as isolates/1000 patient-days. Conclusion A high proportion of admitted patients with COVID-19 received empiric antibiotics. In spite of this, we observed a significant reduction in total antimicrobial consumption and reduced rates of MRO isolation in the post-pandemic period. Disclosures All Authors: No reported disclosures

Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China

Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.

Effect of Poor Nutritional Status and Comorbidities on the Occurrence and Outcome of Pneumonia in Elderly Adults

Background: Malnutrition and comorbidity are two common geriatric syndromes. The pathology of pneumonia is multifactorial, making its diagnosis and management a great challenge. Hospital-acquired pneumonia (HAP) and community-acquired pneumonia (CAP) are two main types of pneumonia. However, the effect of geriatric syndromes on pneumonia and its prognosis have not been clearly explored.Methods: We collected the relevant electronic data of inpatients aged over 65 years and diagnosed with pneumonia in the Geriatrics Department Building of the First Affiliated Hospital with Nanjing Medical University between December 2018 and December 2019, and further divided them into HAP group and CAP group. The correlations of age, age-adjusted Charlson Comorbidity Index (aCCI), basic diseases and nutritional indexes (i.e., albumin, electrolyte, hemoglobin) with pneumonia and prognosis were analyzed. We analyzed the associations between infection prognosis/infection level and age, nutritional status, aCCI and underlying diseases, using linear regression model. The box plot was applied to present infection outcome, and the nomogram was built for predicting infection outcomes. We utilized the heat map to show the associations between nutritional status and infection level/outcome in all infected patients, HAP, and CAP.Results: The final study comprised samples of 669 pneumonia patients divided into HAP group (n = 517) and CAP group (n = 152). In all patients, the infection outcome was negatively correlated with age (P = 0.013). The level of albumin was negatively correlated with infection prognosis in all patients (P = 0.03), and negatively correlated with neutrophil count and CRP (P = 0.008, P < 0.001). ACCI was positively correlated with CRP (P = 0.003). The prognosis was negatively associated with age and albumin level. In the patients with basic dementia/Alzheimer's disease and chronic obstructive pulmonary disease/asthma, the prognosis was worse.Conclusion: There was a correlation between poor nutritional status-related indexes and inflammatory indexes. A poor nutritional status might predict a high risk of pneumonia in elderly adults. Advanced age and comorbidities were risk factors for the occurrence and poor prognosis of pneumonia. Therefore, comorbidities should be well-treated in the elderly with pneumonia.

366 Impact of Haemonchus Contortus on Feed Intake in Katahdin Sheep Selected for Parasite Resistance

Haemonchus contortous (Hc) infections have decreased profitability of flocks worldwide through loss of production efficiency and in severe cases death. Countless research efforts have been made in order to further understand the extensive impact of this gastrointestinal nematode (GIN). Quantifying the impact feeding behavior after Hc infection may prove vital, in understanding internal effect on growing lambs. The aim of this study was to evaluate Katahdin lambs with extensive genetic evaluation for feed efficiency and behavior utilizing a Growsafe™ system. Katahdin ewes from the Virginia Tech Southwest Agriculture Research and Extension Center in Glade Springs Virginia were divergently selected based on fecal egg count (FEC) EBV and mated to rams (n = 8) of similar genotype. A subset of lambs was selected for feedlot evaluation and transported to West Virginia University. Lambs were fed a complete pellet (16% CP) ad libitum while individual intake was measured in pen-fed groups with a GrowSafe™ system. Half of the lambs from each sire FEC EBV group were infected with 5,000 Hc L3 larvae and observed for 6 weeks alongside uninfected contemporaries. Artificial Hc infection resulted in lambs becoming modestly infected with FEC peaking in week 5 at 324 eggs/g. Infection level was not significant between all 4 groups (P < 0.05). Lambs administered H.C. had observably lower feed intake 2 days after infection. Both infected groups returned to normal intake with their control groups shortly after. Feed intake dropped again at day 10 and remained low until day 16 for LoFEC sired lambs and day 20 for HiFEC sired lambs. Offspring of low FEC rams exhibited an ability to return to normal intake levels earlier than lambs with high FEC EBV sires. Further analysis is needed with greater levels of parasite infection to confirm negative FEC EBV have a shorter duration of lower feed intake.

Effects of Agaricus bisporus Mushroom Extract on Honey Bees Infected with Nosema ceranae

Agaricus bisporus water crude extract was tested on honey bees for the first time. The first part of the cage experiment was set for selecting one concentration of the A. bisporus extract. Concentration of 200 µg/g was further tested in the second part of the experiment where bee survival and food consumption were monitored together with Nosema infection level and expression of five genes (abaecin, hymenoptaecin, defensin, apidaecin, and vitellogenin) that were evaluated in bees sampled on days 7 and 15. Survival rate of Nosema-infected bees was significantly greater in groups fed with A. bisporus-enriched syrup compared to those fed with a pure sucrose syrup. Besides, the anti-Nosema effect of A. bisporus extract was greatest when applied from the third day which coincides with the time of infection with N. ceranae. Daily food consumption did not differ between the groups indicating good acceptability and palatability of the extract. A. bisporus extract showed a stimulative effect on four out of five monitored genes. Both anti-Nosema and nutrigenomic effects of A. bisporus extract were observed when supplementation started at the moment of N. ceranae infection or preventively (before or simultaneously with the infection).