Prototyping a Knowledge-Based System to Identify Botanical Extracts for Plant Health in Sub-Saharan Africa.

Replacing synthetic pesticides and antimicrobials with plant-based extracts is a current alternative adopted by traditional and family farmers and many organic farming pioneers. A range of natural extracts are already being marketed for agricultural use, but many other plants are prepared and used empirically. A further range of plant species that could be effective in protecting different crops against pests and diseases in Africa could be culled from the large volume of knowledge available in the scientific literature. To meet this challenge, data on plant uses have been compiled in a knowledge base and a software prototype was developed to navigate this trove of information. The present paper introduces this so-called Knomana Knowledge-Based System, while providing outputs related to Spodoptera frugiperda and Tuta absoluta, two invasive insect species in Africa. In early October 2020, the knowledge base hosted data obtained from 342 documents. From these articles, 11,816 uses-experimental or applied by farmers-were identified in the plant health field. In total, 384 crop pest species are currently reported in the knowledge base, in addition to 1547 botanical species used for crop protection. Future prospects for applying this interdisciplinary output to applications under the One Health approach are presented.

Application of the FTA elute card coupled with visual colorimetric loop-mediated isothermal amplification for the rapid diagnosis of Streptococcus agalactiae in farmed tilapia (Oreochromis niloticus).

A method combining the FTA Elute card and visual colorimetric loop-mediated isothermal amplification (FTA-e/LAMP) was tested to diagnose Streptococcus agalactiae infections in vitro and in vivo. FTA-e/LAMP consists of two main steps: first, the FTA card is used to extract DNA and then a colorimetric loop-mediated isothermal amplification (LAMP) reaction is carried out on the extracted DNA. In vitro sensitivity was 1.9 x 102  CFU/mL, and regarding specificity, all nine S. agalactiae strains tested positive. All Streptococcus spp. tested negative, except for S. dysgalactiae, thereby indicating the need for another set of primers to distinguish this species from S. agalactiae. To diagnose S. agalactiae infections using FTA-e/LAMP in vivo, two experimental trials on juvenile Oreochromis niloticus infected with bovine or piscine strains were carried out. Sensitivity in symptomatic fish was 100%, and 50.7% of fish without signs were positive. All negative control fish tested negative (n = 28). No bacteria were detected after 16 days post-infection (dpi). Accuracy during the first week (1-7 dpi) was 89% and decreased to 44% thereafter (10-22 dpi). FTA-e/LAMP results suggest that this method is a promising tool for early and fast diagnosis of S. agalactiae on tilapia farms.

Aquaculture at the crossroads of global warming and antimicrobial resistance.

In many developing countries, aquaculture is key to ensuring food security for millions of people. It is thus important to measure the full implications of environmental changes on the sustainability of aquaculture. We conduct a double meta-analysis (460 articles) to explore how global warming and antimicrobial resistance (AMR) impact aquaculture. We calculate a Multi-Antibiotic Resistance index (MAR) of aquaculture-related bacteria (11,274 isolates) for 40 countries, of which mostly low- and middle-income countries present high AMR levels. Here we show that aquaculture MAR indices correlate with MAR indices from human clinical bacteria, temperature and countries' climate vulnerability. We also find that infected aquatic animals present higher mortalities at warmer temperatures. Countries most vulnerable to climate change will probably face the highest AMR risks, impacting human health beyond the aquaculture sector, highlighting the need for urgent action. Sustainable solutions to minimise antibiotic use and increase system resilience are therefore needed.

Characterization of Endophytic Streptomyces griseorubens MPT42 and Assessment of Antimicrobial Synergistic Interactions of its Extract and Essential Oil from Host Plant Litsea cubeba.

The present study aimed to evaluate the synergistic effects of the crude ethyl acetate extract (CEAE) from endophytic actinomycete MPT42 and essential oil (EO) of the same host plant Litsea cubeba. The isolate MPT42, exhibiting broad-spectrum antimicrobial activities and harboring all three antibiotic-related biosynthetic genes pks-I, pks-II, and nrps, was identified as Streptomycete griseorubens based on an analysis of the morphology, physiology, and 16S rDNA sequence. Minimum inhibitory concentrations (MICs) and the fractional inhibitory concentration index were used to estimate the synergistic effects of various combined ratios between CEAE or antibiotics (erythromycin, vancomycin) and EO toward 13 microbial strains including pathogens. L. cubeba fruit EO, showing the main chemical constituents of 36.0% citral, 29.6% carveol, and 20.5% limonene, revealed an active-low against tested microbes (MICs ≥ 600 µg/mL). The CEAE of S. griseorubens culture exhibited moderate-strong antimicrobial activities against microbes (MICs = 80-600 µg/mL). Analysis of the mechanism of action of EO on Escherichia coli ATCC 25922 found that bacterial cells were dead after 7 h of the EO treatment at 1 MIC (5.5 mg/mL), where 62% cells were permeabilized after 2 h and 3% of them were filament (length ≥ 6 µm). Combinations of CEAE, erythromycin, or vancomycin with EO led to significant synergistic antimicrobial effects against microbes with 4-16 fold reduction in MIC values when compared to their single use. Interestingly, the vancomycin-EO combinations exhibited a strong synergistic effect against five Gram-negative bacterial species. This could assume that the synergy was possibly due to increasing the cell membrane permeability by the EO acting on the bacterial cells, which allows the uptake and diffusion of antimicrobial substances inside the cell easily. These findings in the present study therefore propose a possible alternative to combat the emergence of multidrug-resistant microbes in veterinary and clinics.

Endophytic Actinobacteria Associated with Dracaena cochinchinensis Lour.: Isolation, Diversity, and Their Cytotoxic Activities.

Dracaena cochinchinensis Lour. is an ethnomedicinally important plant used in traditional Chinese medicine known as dragon's blood. Excessive utilization of the plant for extraction of dragon's blood had resulted in the destruction of the important niche. During a study to provide a sustainable way of utilizing the resources, the endophytic Actinobacteria associated with the plant were explored for potential utilization of their medicinal properties. Three hundred and four endophytic Actinobacteria belonging to the genera Streptomyces, Nocardiopsis, Brevibacterium, Microbacterium, Tsukamurella, Arthrobacter, Brachybacterium, Nocardia, Rhodococcus, Kocuria, Nocardioides, and Pseudonocardia were isolated from different tissues of D. cochinchinensis Lour. Of these, 17 strains having antimicrobial and anthracyclines-producing activities were further selected for screening of antifungal and cytotoxic activities against two human cancer cell lines, MCF-7 and Hep G2. Ten of these selected endophytic Actinobacteria showed antifungal activities against at least one of the fungal pathogens, of which three strains exhibited cytotoxic activities with IC50-values ranging between 3 and 33 µg·mL-1. Frequencies for the presence of biosynthetic genes, polyketide synthase- (PKS-) I, PKS-II, and nonribosomal peptide synthetase (NRPS) among these 17 selected bioactive Actinobacteria were 29.4%, 70.6%, and 23.5%, respectively. The results indicated that the medicinal plant D. cochinchinensis Lour. is a good niche of biologically important metabolites-producing Actinobacteria.

Detection of molecular changes induced by antibiotics in Escherichia coli using vibrational spectroscopy.

This study aimed to test Raman (400-1800cm-1) and Infra-red (1900-500cm-1) spectroscopies followed by statistical analysis (principal component analysis) to detect molecular changes induced by antibiotics (ampicillin, cefotaxime - cell wall synthesis inhibitors, tetracycline - protein synthesis inhibitor, ciprofloxacin - DNA synthesis inhibitor) against Escherichia coli TOP10. In case of ampicillin and cefotaxime, a decrease in protein bands in both Raman (1240, 1660cm-1), and IR spectra (1230, 1530, 1630cm-1), and an increase in carbohydrate bands (1150, 1020cm-1) in IR spectra were observed. Tetracycline addition caused an increase in nucleic acid bands (775, 1478, 1578cm-1), a sharp decrease in phenylalanine (995cm-1) in Raman spectra and the amide I and amide II bands (1630, 1530cm-1) in IR spectra, an increase in DNA in both Raman (1083cm-1) and IR spectra (1080cm-1). Regarding ciprofloxacin, an increase in nucleic acids (775, 1478, 1578cm-1) in Raman spectra and in protein bands (1230, 1520, 1630cm-1), in DNA (1080cm-1) in IR spectra were detected. Clear discrimination of antibiotic-treated samples compared to the control was recorded, showing that Raman and IR spectroscopies, coupled to principal component analysis for data, could be used to detect molecular modifications in bacteria exposed to different classes of antibiotics. These findings contribute to the understanding of the mechanisms of action of antibiotics in bacteria.

Effect of steam and lactic acid treatments on the survival of Salmonella Enteritidis and Campylobacter jejuni inoculated on chicken skin.

Campylobacteriosis and salmonellosis are the most frequently reported zoonotic infectious diseases. The present work evaluated the effectiveness of steam treatment at 100 °C for 8s, a 5% lactic acid treatment for 1 min and their combination for inactivating Salmonella Enteritidis and Campylobacter jejuni inoculated on chicken skin. The impact of each treatment on the total aerobic mesophilic bacteria and the effect of rinsing after contact with lactic acid were also evaluated. Residual bacteria were counted immediately after treatment or after seven days of storage at 4 °C. Results demonstrated the immediate efficiency of the steam and the combined treatments with reductions of approximately 6 and 5 log cfu/cm2 respectively for S. Enteritidis and C. jejuni. They also showed significant reductions (equal to or >3.2 log cfu/cm2) in the total aerobic mesophilic plate count. Lactic acid had a persistent effect on pathogen growth during storage which was significantly higher when the skin was not rinsed, reaching reductions of 3.8 log cfu/cm2 for both S. Enteritidis and C. jejuni. Only the combined treatments significantly reduced the recovery of the total aerobic mesophilic bacteria during storage. The significant reductions in both pathogens and total aerobic mesophilic bacteria on treated chicken skins are possible ways to improve the safety and shelf life of the product although high levels of indigenous non-pathogenic bacteria may be beneficial due to their protective effect against potential re-contamination of chicken skin.

Antimicrobial effects of essential oils of Cinnamosma fragrans on the bacterial communities in the rearing water of Penaeus monodon larvae.

Farmed shrimps are vectors of various Vibrio species that are considered a potential health hazard. Previous study has shown that Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio alginolyticus dominated in the water and larval samples of shrimp hatchery (Randrianarivelo et al. 2010 ). The effects of two essential oils (EOs) of Cinnamosma fragrans, an endemic plant to Madagascar (B8: linalool-type and B143: 1,8-cineole-type), were determined on the total heterotrophic aerobic bacteria and the Vibrio concentrations in the rearing water of Penaeus monodon hatchery. The assays took place in OSO Farming's shrimp hatchery in Madagascar. EOs were directly added to the water tank. The bacterial concentrations of water tank were assessed on marine agar and thiosulfate citrate bile sucrose agar. The larvae culture corresponded to four replicates each of B8, B143, erythromycin (E), and control (oil and antibiotic free). The bacterial concentration of the rearing water in B8, B143, and antibiotic (E) tanks were significantly lower (p < 0.05) than in the control. Further, there was no significant difference (p > 0.05) between the three treatments B8, B143, and E. This study demonstrated that both EOs of C. fragrans, like antibiotic, inhibited bacterial growth in the rearing water of P. monodon larvae. The potential of C. fragrans EO to control the bacterial load in in vivo conditions of P. monodon hatchery makes it a relevant option for producers to minimize risk of Vibrio growth in the rearing water of larvae, which is the primary source of colonization of shrimp larvae.

Effects of steam and lactic acid treatments on inactivation of Listeria innocua surface-inoculated on chicken skins.

Effectiveness of combined steam (10 and 60 s, 70 degrees C and 98 degrees C) and chemical treatments, using concentrated solutions of lactic acid (1 and 30 min, 5% and 10% lactic acid), on the inactivation of Listeria innocua inoculated on the surface of chicken skins have been studied. Surviving bacteria on the skin were enumerated immediately after treatment, and after 7 days of storage at 4 degrees C. The most effective treatment was the combination of steam of 98 degrees C and 10% lactic acid with its immediate efficacy being mainly attributed to the applied heat treatment. However, after 7-day storage, the treatment's effectiveness was mainly due to the applied acid treatment, which prevented growth of the bacteria that survived the heat treatment. Milder treatments (70 degrees C steam, 5% lactic acid) revealed a genuine synergy between the heat and acid treatments, paving the way for an effective means of reducing bacterial load on the surface of poultry without affecting the product's "raw" appearance.

Chemiluminescent and bioluminescent assays as innovative prospects for mycotoxin determination in food and feed.

Mycotoxin contamination of food and feedstuffs is among the top priorities for human and animal safety. The currently used techniques for mycotoxin determination, either chromatography or ELISA, are unsuitable for routine in-field assessment. There is an urgent need for other accurate, simple and cost-effective techniques that can be used as a screening tool for a rapid estimation of mycotoxin contamination in commodity lots. This paper reviews the literature on the use of chemiluminescence (CL) and bioluminescence (BL) assays for direct or indirect mycotoxin assessment. The chemiluminescence immunoassays, adenosine triphosphate (ATP) bioluminescence and bioassays are reviewed and their advantages and limitations discussed. These techniques used in food testing and the pharmaceutical industry offer promise as rapid techniques for mycotoxin determination. Chemiluminescence and bioluminescence bioassays are the most innovative alternatives to the conventional techniques used for mycotoxin determination in food and feed.