Efficacy of Noni (Morinda citrifolia L.) Ethanolic Leaf Extract Against German Cockroach (Blattella germanica L.).

<b>Background and Objective:</b> The ethanol extract of noni leaves (<i>Morinda citrifolia</i> L.) can be used as insecticides to control populations of German cockroaches that have been resistant to synthetic insecticides. This study aimed to determine the potency of the ethanol extract of noni leaves to kill and repel German cockroaches and affect the amount of food consumed. <b>Materials and Methods:</b> The methods used in this study were the contact toxicity test, the repellency test and the food consumption test. The noni leaves extract and German cockroach populations were provided in the laboratory. <b>Results:</b> The noni leaves extract concentration of 20% (residue of 3.14 mg cm² ¹) was very effective in killing the standard population and effective in killing the field population of German cockroaches. The sub-lethal concentration noni leaves extract of 0.36% (residue 0.056 mg cm² ¹) and 1.08% (residue 0.169 mg cm² ¹) was very high grade as repellent of German cockroaches. The sub-lethal concentration of noni leaves extract did not inhibit the amount of food consumption in German cockroach populations. <b>Conclusion:</b> Leaves of noni plants can be used as bioinsecticides to control German cockroach populations that have been resistant to commercial insecticides.

Chloroplast DNA analysis of the invasive weed, Himalayan balsam (Impatiens glandulifera), in the British Isles.

Impatiens glandulifera or Himalayan balsam (HB), is an invasive alien weed throughout the British Isles (BI). Classical biological control of HB in the BI using a rust fungus from the Himalayan native range was implemented in 2014. However, not all HB populations are susceptible to the two rust strains currently released. Additional strains are needed that infect resistant populations in order to achieve successful control. These are best sourced from the historical collecting sites. A molecular analysis was conducted using six chloroplast DNA sequences from leaf material from across the BI and the native range. Herbarium samples collected in the Himalayas between 1881 and 1956 were also included. Phylogenetic analyses resulted in the separation of two distinct groups, one containing samples from the BI and the native range, and the other from the BI only; suggesting that HB was introduced into the BI on at least two occasions. The former group is composed of two subgroups, indicating a third introduction. Ten and 15 haplotypes were found in the introduced and native range respectively, and with two of these found in both regions. Results show where to focus future surveys in the native range to find more compatible rust strains.

Advances in the control of phytopathogenic fungi that infect crops through their root system.

Productivity and economic sustainability of many herbaceous and woody crops are seriously threatened by numerous phytopathogenic fungi. While symptoms associated with phytopathogenic fungal infections of aerial parts (leaves, stems and fruits) are easily observable and therefore recognizable, allowing rapid or preventive action to control this type of infection, the effects produced by soil-borne fungi that infect plants through their root system are more difficult to detect. The fact that these fungi initiate infection and damage underground implies that the first symptoms are not as easily noticeable, and therefore both crop yield and plant survival are frequently severely compromised by the time the infection is found. In this paper we will review and discuss recent insights into plant-microbiota interactions in the root system crucial to understanding the beginning of the infectious process. We will also review different methods for diminishing and controlling the infection rate by phytopathogenic fungi penetrating through the root system including both the traditional use of biocontrol agents such as antifungal compounds as well as some new strategies that could be used because of their effective application, such as nanoparticles, virus-based nanopesticides, or inoculation of plant material with selected endophytes. We will also review the possibility of modeling and influencing the composition of the microbial population in the rhizosphere environment as a strategy for nudging the plant-microbiome interactions toward enhanced beneficial outcomes for the plant, such as controlling the infectious process.

Soil fungi enable the control of gastrointestinal nematodes in wild bovidae captive in a zoological park: a 4-year trial.

The control of gastrointestinal nematodes among ruminants maintained in zoological parks remains difficult due to infective stages develop in the soil. For the purpose to improve the possibilities of the control of gastrointestinal nematodes (genera Trichostrongylus, Nematodirus, Chabertia and Haemonchus) affecting wild captive bovidae ruminants belonging to the subfamilies Antilopinae, Caprinae, Bovinae and Reduncinae, commercial pelleted feed enriched with a blend of 104-105 spores of both filamentous fungi Mucor circinelloides + Duddingtonia flagrans per kg meal was provided for a period of 3.5 years. All animals were dewormed at the beginning of the trial and also when exceeding a cut-off point of 300 eggs per gram of feces (EPG). The anthelmintic efficacy ranged between 96 and 100%. The need for repeating the administration of parasiticide treatment disappeared at the 24th month of study in the Antilopinae individuals, and at the 8th month in the Caprinae, Bovinae and Reduncinae. No side-effects were observed on the skin or in the digestive, respiratory or reproductive system. It was concluded that this strategy provides a sustainable tool for preventing the contamination of paddocks where captive ruminants are maintained, decreasing the risk of infection by gastrointestinal nematodes and consequently the need of frequent deworming.

Lactobacillus salivarius A3iob Reduces the Incidence of Varroa destructor and Nosema Spp. in Commercial Apiaries Located in the Northwest of Argentina.

Lactobacillus salivarius A3iob was administered to productive colonies belonging to commercial apiaries of small beekeepers (around 30-50 hives each one), from four departments of the province of Jujuy (Argentina): Yala, Tilquiza, El Carmen, and Los Alisos. The incidence of Varroa destructor and Nosema spp., before and after winter, was monitored during 2 years of study (2014-2015). Depending on the geographical location of each apiary and the application time, a monthly dose of the bacteria (105 CFU/mL) reduced the levels of varroasis between 50 and 80%. Interestingly, L. salivarius A3iob cells remitted the percentage of the mites to undetectable values in an apiary treated with flumethrin (at Yala, Yungas region).On the other hand, the spore levels of Nosema spp. in the lactobacilli-treated colonies also depended on the apiary and the year of application, but a significant decrease was mainly observed in the post-winter period. However, at Rivera (El Carmen's department), no significant changes were detected in both parameters.These results obtained after 2 years of work suggest that delivering L. salivarius A3iob cells to the bee colonies can become a new eco-friendly tool to cooperate with the control of these bees' pests.

Potential Use of Biotherapeutic Bacteria to Target Colorectal Cancer-Associated Taxa.

The role of the gut microbiome in human health and disease is the focus of much attention. It has been widely agreed upon that our gut bacteria play a role in host immunity, nutrient absorption, digestion, metabolism, and other key drivers of health. Furthermore, certain microbial signatures and specific taxa have also been associated with the development of diseases, such as obesity; inflammatory bowel disease; and, indeed, colorectal cancer (CRC), which is the focus of this review. By extension, such taxa represent potential therapeutic targets. In particular, the emerging human pathogen Fusobacterium nucleatum represents an important agent in CRC development and its control within the gastrointestinal tract is desirable. This paper reviews the principal bacterial pathogens that have been associated with CRC to date and discusses the in vitro and human studies that have shown the potential use of biotherapeutic strains as a means of targeting CRC-associated bacteria.

Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of nematodiosis in extensive bovine production system.

Cattle production in extensive systems favours the occurrence of gastrointestinal nematodes, and the use of nematophagous fungi complements the control strategies for these nematodes. The aim of this study was to evaluate the effectiveness of the fungi Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of gastrointestinal parasitic nematodes in grazing cattle. Twenty-four calves were randomly divided into four groups and allocated to independent paddocks from February 2018 to January 2019. In the first group, the animals received pellets containing P. chlamydosporia. In the second group, the animals received pellets containing A. cladodes. In the third group, the animals received pellets containing a combination of the fungi A. cladodes and P. chlamydosporia. In the control group, the animals received pellets without fungus. The combined use of A. cladodes and P. chlamydosporia showed greater efficacy in the biological control of bovine gastrointestinal parasitic nematodes than the same fungi used separately. The parasite load was lower and weight gain was greater (P ⩽ 0.05) in the groups of cattle treated with nematophagous fungi. Therefore, the use of A. cladodes and P. chlamydosporia is promising in the biological control of nematodiosis in cattle.

The impact of RHDV-K5 on rabbit populations in Australia: an evaluation of citizen science surveys to monitor rabbit abundance.

The increasing popularity of citizen science in ecological research has created opportunities for data collection from large teams of observers that are widely dispersed. We established a citizen science program to complement the release of a new variant of the rabbit biological control agent, rabbit haemorrhagic disease virus (RHDV), known colloquially as K5, across Australia. We evaluated the impact of K5 on the national rabbit population and compared citizen science and professionally-collected spotlight count data. Of the citizen science sites (n = 219), 93% indicated a decrease in rabbit abundance following the release of K5. The overall finite monthly growth rate in rabbit abundance was estimated as 0.66 (95%CI, 0.26, 1.03), averaging a monthly reduction of 34% at the citizen science sites one month after the release. No such declines were observed at the professionally monitored sites (n = 22). The citizen science data submissions may have been unconsciously biased or the number of professional sites may have been insufficient to detect a change. Citizen science participation also declined by 56% over the post-release period. Future programs should ensure the use of blinded trials to check for unconscious bias and consider how incentives and/or the good will of the participants can be maintained throughout the program.

Infection model for analyzing biological control of coffee rust using bacterial anti-fungal compounds.

Coffee rust is one of the main diseases that affect coffee plantations worldwide (Cressey, 2013 [10]). This causes an important economic impact in the coffee production industry in countries where coffee is an important part of the economy. A common method for combating this disease is using copper hydroxide as a fungicide, which can have damaging effects both on the coffee tree and on human health (Haddad et al., 2013 [13]). A novel method for biological control of coffee rust using bacteria has been proven to be an effective alternative to copper hydroxide fungicides as anti-fungal compounds (Haddad et al., 2009 [12]). In this paper, we develop and explore a spatial stochastic model for this interaction in a coffee plantation. We analyze equilibria for specific control strategies, as well as compute the basic reproductive number, R0, of individual coffee trees, conditions for local and global stability under specific conditions, parameter estimation of key parameters, as well as sensitivity analysis, and numerical experiments under local and global control strategies for key scenarios.

In vitro efficacy of Duddingtonia flagrans against nematodes of sheep based on in vivo calculations / Eficácia in vitro de Duddingtonia flagrans contra nematoides de ovinos com base em cálculo in vivo

Abstract Duddingtonia flagrans has been tested as an alternative parasite control, but data from in vitro experiments based on in vivo calculations describing nematophagous fungi predation in nematodes are restricted. The objective of this work was to determine the efficacy of D. flagrans against sheep nematode larvae in vitro using in vivo calculations. Fecal samples were introduced to fungi in different concentrations: 0.0/control; 0.05; 0.1; 0.2; 0.4; 0.8; 1.6; 3.2; and 6.4 g corresponding, respectively, to 583.000; 1.166.000; 2.332.000; 4.664.000; 9.328.000; 18.656.000; 37.312.000 and 74.624.000 chlamydospores/kg of body weight. The material was incubated for 14 days, before the larvae recovery (Assay 1). Assay 2 was carried out with the doses of 0.00625; 0.0125; and 0.025 g. The results showed a negative correlation between fungal concentrations and larval numbers for both assays. The fungus demonstrated an efficacy above 89% in both assays. Thus, we consider that the data from in vitro studies based on in vivo calculations may optimize the fungi quantities for field experiments.

Resumo Duddingtonia flagrans tem sido testado como uma alternativa no controle de parasitos, entretanto, trabalhos in vitro da predação de nematoides por fungos nematófagos correlacionados com cálculos baseados para testes in vivo são restritos. O objetivo deste trabalho foi determinar a eficácia in vitro de D. flagrans contra larvas de nematoides de ovinos tendo como base cálculos in vivo. Amostras fecais receberam a adição do fungo em diferentes concentrações: 0.0/controle; 0,05; 0,1; 0,2; 0,4; 0,8; 1,6; 3,2 e 6,4 gramas correspondendo, respectivamente, às seguintes dosagens: 583.000; 1.166.000; 2.332.000; 4.664.000; 9.328.000; 18.656.000; 37.312.000 e 74.624.000 clamidósporos/Kg de peso vivo animal. O material foi incubado por 14 dias, para recuperação das larvas (Ensaio 1). O Ensaio 2 foi realizado com concentrações de 0,00625; 0,0125 e 0,025 g. Foi observada correlação negativa entre a concentração fúngica e o número de larvas, nos dois ensaios. O fungo demonstrou eficácia acima de 89% em ambos os ensaios. A partir destes dados, acreditamos que ensaios in vitro baseados em cálculos in vivo podem aprimorar as dosagens para a realização de experimentos a campo.

In vitro efficacy of Duddingtonia flagrans against nematodes of sheep based on in vivo calculations.

Duddingtonia flagrans has been tested as an alternative parasite control, but data from in vitro experiments based on in vivo calculations describing nematophagous fungi predation in nematodes are restricted. The objective of this work was to determine the efficacy of D. flagrans against sheep nematode larvae in vitro using in vivo calculations. Fecal samples were introduced to fungi in different concentrations: 0.0/control; 0.05; 0.1; 0.2; 0.4; 0.8; 1.6; 3.2; and 6.4 g corresponding, respectively, to 583.000; 1.166.000; 2.332.000; 4.664.000; 9.328.000; 18.656.000; 37.312.000 and 74.624.000 chlamydospores/kg of body weight. The material was incubated for 14 days, before the larvae recovery (Assay 1). Assay 2 was carried out with the doses of 0.00625; 0.0125; and 0.025 g. The results showed a negative correlation between fungal concentrations and larval numbers for both assays. The fungus demonstrated an efficacy above 89% in both assays. Thus, we consider that the data from in vitro studies based on in vivo calculations may optimize the fungi quantities for field experiments.

Huanglongbing Control: Perhaps the End of the Beginning.

Huanglongbing (HLB) is one of the most destructive citrus plant diseases worldwide. It is associated with the fastidious phloem-limited α-proteobacteria 'Candidatus Liberibacter asiaticus', 'Ca. Liberibacter africanus' and 'Ca. Liberibacter americanus'. In recent years, HLB-associated Liberibacters have extended to North and South America. The causal agents of HLB have been putatively identified, and their transmission pathways and worldwide population structure have been extensively studied. However, very little is known about the epidemiologic relationships of Ca. L. asiaticus, which has limited the scope of HLB research and especially the development of control strategies. HLB-affected plants produce damaged fruits and die within several years. To control the disease, scientists have developed new compounds and screened existing compounds for their antibiotic and antimicrobial activities against the disease. These compounds, however, have very little or even no effect on the disease. The aim of the present review was to compile and compare different methods of HLB disease control with newly developed integrative strategies. In light of recent studies, we also describe how to control the vectors of this disease and the biological control of other citrus plant pathogens. This work could steer the attention of scientists towards integrative control strategies.

Sigma factor RpoN employs a dual transcriptional regulation for controlling twitching motility and biofilm formation in Lysobacter enzymogenes OH11.

Lysobacter is a Gram-negative genus comprising a group of environmental bacteria with abilities to produce abundant novel antibiotics, as well as adopting a unique type IV pilus (T4P)-mediated twitching motility (TM) that remains poorly understood. Here, we employ L. enzymogenes OH11 exhibiting significant antifungal activity as a working model to address this issue. Via mutating the 28 potential sigma factors in strain OH11, we have identified one protein RpoNOH11 (sigma 54) that is indispensable for T4P formation and TM. We further showed that RpoNOH11 not only regulates the transcription of pilA, but also another crucial gene chpA that encodes a hybrid two-component transduction system. The L. enzymogenes RpoNOH11 was found to directly bind to the promoter of chpA to control its transcription, which is found to be essential for the T4P-mediated TM. To our knowledge, such a transcriptional regulation performed by RpoN in control of bacterial TM has never been reported. Finally, we showed that L. enzymogenes OH11 could also produce biofilm that is likely employed by this strain to infect fungal pathogens. Mutation of rpoN OH11, pilA and chpA all led to a significant decrease in biofilm formation, suggesting that the dual transcriptional regulation of pilA and chpA by RpoNOH11 plays a key role for RpoNOH11 to modulate the biofilm formation in L. enzymogenes. Overall, this study identified chpA as a new target of RpoN for controlling the T4P-mediated twitching motility and biofilm formation in L. enzymogenes OH11.

Microbiota, Liver Diseases, and Alcohol.

Being overweight and obesity are the leading causes of liver disease in Western countries. Liver damage induced by being overweight can range from steatosis, harmless in its simple form, to steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. Alcohol consumption is an additional major cause of liver disease. Not all individuals who are overweight or excessively consume alcohol develop nonalcoholic fatty liver diseases (NAFLD) or alcoholic liver disease (ALD) and advanced liver disease. The role of the intestinal microbiota (IM) in the susceptibility to liver disease in this context has been the subject of recent studies. ALD and NAFLD appear to be influenced by the composition of the IM, and dysbiosis is associated with ALD and NAFLD in rodent models and human patient cohorts. Several microbial metabolites, such as short-chain fatty acids and bile acids, are specifically associated with dysbiosis. Recent studies have highlighted the causal role of the IM in the development of liver diseases, and the use of probiotics or prebiotics improves some parameters associated with liver disease. Several studies have made progress in deciphering the mechanisms associated with the modulation of the IM. These data have demonstrated the intimate relationship between the IM and metabolic liver disease, suggesting that targeting the gut microbiota could be a new preventive or therapeutic strategy for these diseases.

Bacteriophage Clinical Use as Antibacterial "Drugs": Utility and Precedent.

For phage therapy-the treatment of bacterial infections using bacterial viruses-a key issue is the conflict between apparent ease of clinical application, on the one hand, and on the other hand, numerous difficulties that can be associated with undertaking preclinical development. These conflicts between achieving efficacy in the real world versus rigorously understanding that efficacy should not be surprising because equivalent conflicts have been observed in applied biology for millennia: exploiting the inherent, holistic tendencies of useful systems, e.g., of dairy cows, inevitably is easier than modeling those systems or maintaining effectiveness while reducing such systems to isolated parts. Trial and error alone, in other words, can be a powerful means toward technological development. Undertaking trial and error-based programs, especially in the clinic, nonetheless is highly dependent on those technologies possessing both inherent safety and intrinsic tendencies toward effectiveness, but in this modern era we tend to forget that ideally there would exist antibacterials which could be thus developed, that is, with tendencies toward both safety and effectiveness, and which are even relatively inexpensive. Consequently, we tend to demand rigor as well as expense of development even to the point of potentially squandering such utility, were it to exist. In this review I lay out evidence that in phage therapy such potential, in fact, does exist. Advancement of phage therapy unquestionably requires effective regulation as well as rigorous demonstration of efficacy, but after nearly 100 years of clinical practice, perhaps not as much emphasis on strictly laboratory-based proof of principle.

Ecological Therapeutic Opportunities for Oral Diseases.

The three main oral diseases of humans, that is, caries, periodontal diseases, and oral candidiasis, are associated with microbiome shifts initiated by changes in the oral environment and/or decreased effectiveness of mucosal immune surveillance. In this review, we discuss the role that microbial-based therapies may have in the control of these conditions. Most investigations on the use of microorganisms for management of oral disease have been conducted with probiotic strains with some positive but very discrete clinical outcomes. Other strategies such as whole oral microbiome transplantation or modification of community function by enrichment with health-promoting indigenous oral strains may offer more promise, but research in this field is still in its infancy. Any microbial-based therapeutics for oral conditions, however, are likely to be only one component within a holistic preventive strategy that should also aim at modification of the environmental influences responsible for the initiation and perpetuation of microbiome shifts associated with oral dysbiosis.

Control of Haemonchus contortus in sheep using basidiocarps of Agaricus blazei Murril.

OBJECTIVE: This study evaluated the effects in vitro and in vivo of Agaricus blazei against Haemonchus contortus in sheep. METHODS: The in vitro efficacy of aqueous extract on egg hatching inhibition (EHI) was investigated and after 72 h incubation with varying concentrations the effects on, blastomeres, embryonated eggs, and first stage larvae (L1) were evaluated. Larval development inhibition (LDI) for dry powder and the aqueous extract were evaluated in fecal cultures of sheep infected with H. contortus. In vivo efficacy was determined by reduction in fecal egg count (FEC). Lambs were treated with powder A. blazei (11.4 g/kg pc) or trichlorfon, or were untreated and the possible toxicity of this fungus was monitored by plasmatic enzyme analysis. RESULTS: Concentrations equal to and higher than 3.62 mg/mL and of aqueous extract were 100% effective in the EHI test. In the LDI test, LC90 was estimated for 5.66 and 106.0 mg/g fecal culture for aqueous extract and powder, respectively. The mean FEC in lambs 14 days post-treatment with A. blazei powder was significantly lower than observed for the negative control, and the serum levels of aspartate transaminase and alanine transaminase were normal. CONCLUSION: The fungi supplementation promotes, respectively, high and moderate anthelmintic efficacy with in vitro and in vivo tests, respectively, suggesting it as an alternative or complementary treatment for haemonchosis in sheep.

A minireview on the in vitro and in vivo experiments with anti-Escherichia coli O157:H7 phages as potential biocontrol and phage therapy agents.

Phage therapy is an old method of combating bacterial pathogens that has recently been taken into consideration due to the alarming spread of antibiotic resistance. Escherichia coli O157:H7 is a foodborne pathogen that causes hemorrhagic colitis and life-threatening Hemolytic Uremic Syndrome (HUS). There are several studies on isolation of specific phages against E. coli O157:H7 and more than 60 specific phages have been published so far. Although in vitro experiments have been successful in elimination or reduction of E. coli O157:H7numbers, in vivo experiments have not been as promising. This may be due to escape of bacteria to locations where phages have difficulties to enter or due to the adverse conditions in the gastrointestinal tract that affect phage viability and proliferation. To get around the latter obstacle, an alternative phage delivery method such as polymer microencapsulation should be tried. While the present time results are not very encouraging the work should be continued as more efficient phage treatment regimens might be found in future.

The re-emerging arboviral threat: Hidden enemies: The emergence of obscure arboviral diseases, and the potential use of Wolbachia in their control.

Mayaro, Oropouche, and O'Nyong-Nyong share many traits with more prominent arboviruses, like dengue and yellow fever, chikungunya, and Zika. These include severe clinical symptoms, multiple animal hosts, and widespread vector species living in close proximity to human habitats, all of which constitute significant risk factors for more frequent outbreaks in the future, greatly increasing the potential of these hidden enemies to follow Zika and become the next wave of global arboviral threats. Critically, the current dearth of knowledge on these arboviruses might impede the success of future control efforts, including the potential application of Wolbachia pipientis. This bacterium inherently possesses broad anti-pathogen properties and a means of genetic drive that allows it to eliminate or replace target vector populations. We conclude that control of obscure arboviruses with Wolbachia might be possible, but successful implementation will be critically dependent on the ability to transinfect key vector species.

The potassium impermeable apical membrane of insect epithelia: a target for development of safe pesticides

Columnar cell apical membranes (CCAM) in series with goblet cell apical membranes (GCAM) form an electroosmotic barrier separating the midgut lumen from epithelial cell cytoplasm. A unique K+ ATPase in GCAM generates three gradients across this barrier. A greater than 180 mV electrical gradient (lumen positive) drives amino acid uptake through voltage-dependent K+ symports. A greater than 1000-fold [H+] gradient (lumen alkaline) and a greater than 10-fold [K+] gradient (lumen concentrated) are adaptations to the high tannin and high K+ content, respectively, in dietary plant material. Agents which act on the apical membrane and disrupt the PD, H+, or K+ gradients are potential insecticides. Insect sensory epithelia and mammalian stria vascularis maintain similar PD and K+ gradients but would not be exposed to ingested anti-apical membrane insecticides. Following the demonstration by Sacchi et al. that Bacillus thuringiensis delta-endotoxin (Bt) induces specifically a K+ conductance increase in CCAM vesicles, we find that the K+ channel blocking agent, Ba2+, completely reverses Bt inhibition of the K+-carried short circuit current in the isolated midgut of Manduca sexta. Progress in characterizing the apical membrane includes finding that fluorosulfonylbenzoyladenosine binds specifically to certain GCAM polypeptides and that CCAM vesicles can be mass produced by Ca2+ or Mg2+ precipitation from Manduca sexta midgut.