Characterization of an Insoluble and Soluble Form of Melanin Produced by Streptomyces cavourensis SV 21, a Sea Cucumber Associated Bacterium

Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg²⁺ ions and the insoluble PPM is dominated by Ca²⁺ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.     

Meta-analysis of fungicide efficacy on soybean target spot and cost–benefit assessment

Target spot of soybean has spread in Brazil, the southeastern United States and Argentina in the last decade. A collaborative network of field Uniform Fungicide Trials (UFT) in Brazil was created in 2011 to study the target spot control efficacy of fungicides, including azoxystrobin + benzovindiflupyr (AZ_BF), carbendazim (CZM), fluxapyroxad + pyraclostrobin (FLUX_PYRA), epoxiconazole + FLUX_PYRA (EPO_FLUX_PYRA), mancozeb (MZB) and prothioconazole + trifloxystrobin (PROT_TRIF). Network meta-analysis was used to conduct a quantitative synthesis of UFT data collected from 2012 to 2016 and to evaluate the effects of disease pressure (DP, low ≤ 35% target spot severity in the nontreated control < high) and year of experiment on the overall mean efficacy and yield response to each of the tested fungicides. Based on mean percentage control of target spot severity, the tested fungicides fall into three efficacy groups (EG): high EG, FLUX_PYRA (76.2% control relative to the nontreated control) and EPO_FLUX_PYRA (75.7% control); intermediate EG, PROT_TRIF (66.5% control) and low EG, MZB (49.6% control), AZ_BF (46.7% control) and CZM (32.4% control). DP had a significant effect on yield response. At DP_Low, the highest response was due to PROT_TRIF (+342 kg ha⁻¹, +12.8%) and EPO_FLUX_PYRA (+295.5 kg ha⁻¹, +11.2%), whereas at DP_High, EPO_FLUX_PYRA and FLUX_PYRA outperformed the other treatments, with yield responses of 503 kg ha⁻¹ (+20.2%) and 469 kg ha⁻¹ (+19.1%), respectively. The probability of a positive return on fungicide investment ranged from 0.26 to 0.56 at DP_Low and from 0.34 to 0.66 at DP_High.     

Spatial patterns of the Citrus leprosis virus and its associated mite vector in systems without intervention

Leprosis is caused by the Citrus leprosis virus cytoplasmic type and is vectored by the mite Brevipalpus yothersi. Miticide applications, which cost $54 million annually, are based on inspection for the presence of mites. The aim of the present study was to characterize the spatial patterns of B. yothersi-infested trees and trees with leprosis symptoms for further improvement in sampling and disease control. The presence of mites and the occurrence of leprosis were assessed over two years in 1160 Valencia trees and 720 Natal trees in a commercial sweet orange grove in Sao Paulo State, Brazil. To assess the natural growth and dispersal of mites and leprosis, mite populations were not controlled during the experimental period. Maps of mite-infested trees and trees with leprosis symptoms were analysed at three different levels of spatial hierarchy using complementary methods, i.e. among adjacent trees within and across rows, within quadrats, and the strength and orientation of aggregation among quadrats. The study showed that the spatial patterns of virus-infected and mite-infested trees were different, with a strong aggregation pattern of trees with leprosis symptoms that increased over time. Conversely, the spatial pattern of B. yothersi showed randomness or weak aggregation at all three spatial hierarchical levels. Disease incidence increased steadily in plots of both cultivars, unlike in mite-infested trees where incidence fluctuated over time. These results have important implications for the development of better management strategies for leprosis. Sampling methods and action thresholds for mite control should consider primary disease inoculum in addition to the incidence of mites.     

Trichoderma species associated with green mould disease of Pleurotus ostreatus and their sensitivity to prochloraz

Green mould disease causes serious economic losses in Pleurotus ostreatus crops worldwide, including in Italy, where prochloraz is the only chemical fungicide allowed to control the disease. The effectiveness of the doses 0.01, 0.05, 0.25 and 1.25 μL L⁻¹ (field dose) of prochloraz (Sponix Flow, 450 g L⁻¹), against colony growth rate and spore germination of Trichoderma pleuroti, T. pleuroticola and T. guizhouense strains on wheat straw extract agar plates were evaluated. Complete inhibition of Trichoderma pleuroti and T. pleuroticola growth was shown by the field dose of prochloraz, and also by the 0.25 μL L⁻¹ dose for T. pleuroti. Complete inhibition of spore germination occurred for all Trichoderma strains at field dose, and at 0.25 μL L⁻¹ for T. pleuroti strains. In in vivo assays, the effect of prochloraz doses 0.05, 0.25 and 1.25 μL L⁻¹ on colonization of straw substrate by T. pleuroti, T. pleuroticola and T. guizhouense inoculated at two spore densities (10² and 10⁵ spores mL⁻¹) immediately after P. ostreatus spawn was studied. Trichoderma pleuroti and T. pleuroticola were both responsible for green mould disease, whereas T. guizhouense was not pathogenic. Trichoderma pleuroti was more aggressive than T. pleuroticola. Prochloraz was effective against T. pleuroti at the field dose, and against T. pleuroticola at 0.25 and 1.25 μL L⁻¹. The study on Trichoderma × Pleurotus interaction type showed that Trichoderma species were active against the mycelial growth of P. ostreatus by competition for space and nutrients, and neither hyphal interaction nor effect by volatile or nonvolatile metabolites occurred.     

Site‐specific management is crucial to managing Mikania micrantha

Increasingly, weeds have been taking on global distributions. With the proliferation of invasive weeds has come the challenge of managing these species over broad geographical regions, with diverse habitats and political jurisdictions. Here, we review the management of Mikania micrantha Kunth (Asteraceae; mile‐a‐minute) throughout its invaded range, extending through most of the Pacific islands and southern and south‐east Asia. Context matters when determining the best course of action for managing M. micrantha, as it has invaded a large variety of agricultural and natural systems. In Queensland, Australia and Florida, USA, M. micrantha has been targeted in relatively successful eradication campaigns, highlighting the importance of early detection and rapid response methods, while elsewhere in its invaded range, populations are either still increasing or showing limited signs of decline. An inter‐regional approach to research and management should incorporate successful management strategies employed throughout the invaded range including, but not limited to, chemical and cultural control practices, manual and mechanical control, classical biological control using the rust fungus Puccinia spegazzinii, plant–plant competition and integrated approaches utilising two or more control methods concurrently. Additional knowledge of M. micrantha genetics is required to determine if management approaches could be fine‐tuned for particular populations. Countries bordering the Mekong River formed a network in 2011 to co‐ordinate the management of invasive species such as M. micrantha. Expanding such a collaborative approach to other regions could further reduce populations of M. micrantha and limit its spread.     

Outcomes of esophageal and gastric bone foreign bodies in dogs

BackgroundBone foreign bodies are commonly encountered in small animal practice. Esophageal bone foreign bodies (E‐bFBs) warrant removal, whereas gastric bone foreign bodies might not.ObjectivesDescribe management and outcomes for dogs with esophageal or gastric bone foreign bodies.AnimalsOne hundred twenty‐nine dogs with esophageal (n = 45) or gastric (n = 84) bone foreign bodies.MethodsRetrospective review of medical records.ResultsDogs with E‐bFBs were younger than dogs with gastric bone foreign bodies (median age esophageal, 4 years [IQR 2‐8]; median age gastric, 6 years [IQR 3‐10]; P = .03), and had a higher bone cross‐sectional area relative to body weight (median esophageal, 98.21 mm²/kg [IQR 48.25‐142.6]; median gastric, 28.6 mm²/kg [IQR 17.25‐64.28]; P < .001). Forty‐two of 45 esophageal foreign bodies were resolved non‐surgically and 3 by esophagotomy. Esophageal erosions were more likely with distal entrapment (OR 12.88, [95% CI 31.95‐129.29], P = .01) and longer duration (OR 18.82 [95% CI 2.22‐273.97], P = .01). Sixty‐two of 84 bone gastric foreign bodies were left in situ. Endoscopic removal was successful in 20 of 22 (91%; 95% CI 70‐99) attempts.Conclusions and Clinical ImportanceWhile all E‐bFBs were dislodged either by advancement into the stomach, endoscopic removal, or esophagotomy, the majority of gastric bone foreign bodies were left in situ for dissolution, with no reported complications. Gastric advancement of E‐bFBs should be considered when oral removal is not feasible, and dissolution can be considered even with large bones.     

Pathology of red spot disease in sea mullet, Mugil cephalus L., from eastern Australia

Abstract. Lesions present on sea mullet, Mugil cephalus L., during the first 3 weeks of red spot disease outbreaks are described. Necrotizing dermatitis is a severe, locally extensive, granulomatous lesion associated with invasion of dermis and underlying skeletal muscle by numerous, non-septate, fungal hyphae 12–18 μm in diameter. Erythematous dermatitis is a mild to severe, focal, chronic active dermatitis without fungal involvement. Lesions intermediate between these two forms, with small to moderate numbers of fungal hyphae in dermis and skeletal muscle also occur. Findings indicate that erythematous dermatitis lesions and intermediate-type lesions subsequently resolve, while necrotizing dermatitis lesions consistently develop into dermal ulcers, with associated severe necrotizing granulomatous myositis. Generally, dermal ulcers occur significantly more often on posterior and dorsal areas of the body surface than on anterior and ventral areas. Lesions heal by combinations of epidermal and dermal repair, fibroplasia, destruction of fungi, removal of necrotic skeletal muscle and regeneration of myofibres. Atrophy of exocrine pancreas occurs in both diseased and clinically normal fish, but is generally more severe in diseased fish. The possible roles of suspected disease determinants, including falls in dissolved oxygen concentrations to sub-lethal levels prior to red spot disease outbreaks, are discussed.     

First isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata (Moreau)

The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed.     

Inhibitory effect of 2,4-dibromophenol and 2,4,6-tribromophenol on larval survival and metamorphosis of the sea urchin <Emphasis Type="Italic">Strongylocentrotus nudus</Emphasis>

ABSTRACT:   As a possible factor leading to the low recruitment level of sea urchins in kelp forests, the inhibitory effect of 2,4-dibromophenol (DBP) and 2,4,6-tribromophenol (TBP) released from the large perennial brown algae Ecklonia kurome and Eisenia bicyclis on survival and metamorphosis of eight-armed larvae of the sea urchin Strongylocentrotus nudus was examined. The percentage of larvae that underwent metamorphosis in filtered sea water after 1 h exposure to one-half dilution of saturated dibromomethane solution (∼60 ppm) as a chemical inducer reached approximately 100% after 1 h, while that in filtered sea water containing 1 ppm TBP was reduced to 73%. This was further reduced to less than 40% in the presence of 10 and 20 ppm TBP after 2 h. In filtered sea water containing 1 and 10 ppm DBP, the proportion of metamorphosed larvae was reduced markedly to 43 and 5% after 2 h, respectively. All larvae exposed to 50 ppm TBP and to 20 and 50 ppm DBP died after 1 h. These findings suggest that DBP is more toxic than TBP for sea urchin larvae, strongly inhibiting their metamorphosis.     

High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.