Early stages of Margaritifera margaritifera glochidiosis in Atlantic salmon: Morphopathological characterization

Freshwater mussels of the order Unionida encyst into the fish mucosa to metamorphose and complete their life cycle, causing a parasitic disease known as glochidiosis. This parasitic stage represents a bottleneck for the survival of naiads, particularly for critically endangered species as Margaritifera margaritifera; however, little is known about the events occurring during this critical stage. Therefore, this study aimed to histologically characterize the development of M. margaritifera glochidiosis in Atlantic salmon to get insight into the pathogenesis of this interaction. Fish exposed to glochidia were sampled during the first 44 days post-exposure, and organs were observed by stereomicroscopy and light microscopy. Glochidia attached to the gills by pinching the lamellar epithelium, whereupon an acute proliferative branchitis engulfed most of the larvae. However, during the first 14 days, a severe detachment of unviable glochidia occurred, associated with the presence of pleomorphic inflammatory infiltrate and epithelial degeneration. In the cases where larvae remained attached, a chronification of the lesions with none to scarce inflammation was observed. These results provide key information to better understand the complex host–parasite interaction during the early stages of glochidiosis and provide valuable information to optimize artificial rearing of naiads in conservation of threatened freshwater mussel populations.     

Prevalence,intensity and histopathology of Zeylanicobdella arugamensis infestation on groupers reared on different aquaculture systems

The marine leech, Zeylanicobdella arugamensis, is a major threat to aquaculture in grouper-producing countries including Indonesia. This study aimed at investigating prevalence, intensity and histopathology of the ectoparasite in humpback and hybrid groupers cultured in different rearing systems. A total of 260 groupers (60 humpback groupers and 200 hybrid groupers) were used for samples. The marine leech was observed on skin, fins, gills and mouth, followed by histopathological assay on the skin tissue. The results showed that prevalence of the leech in both groupers was higher when they were cultured in the floating net cages compared with the hatchery, p < .05. Furthermore, humpback grouper had a higher prevalence than hybrid grouper when they were cultured in a similar system, p < .05. Meanwhile, there was no significant difference in intensity between the two groupers, p > .05. Within the hybrid groupers, the highest prevalence was obtained from hybrid groupers reared in the earthen ponds. Histopathological studies showed that the infected groupers exhibited inflammation, congestion and erosion of the epidermis layer. Hybrid grouper had more severe histopathological lesions in the skin tissues. These results suggested that species and type of aquaculture system had significantly determined the prevalence, intensity and severity of lesion in Z. arugamensis infestation.     

Habitat associations of the Threatened pugnose minnow (Opsopoeodus emiliae) at the northern edge of the species range

Pugnose minnow (Opsopoeodus emiliae) is a small, reclusive species that is widespread in North America, but is one of the rarest fishes in Canada, found in less than 12 known localities in southwestern Ontario. In contrast to most pugnose minnow populations across the global range, Canadian populations are primarily found in turbid systems, potentially indicating persistence in suboptimal conditions. We used data from a multi-gear species and habitat survey in the Canard River, Ontario, a system dominated by agricultural inputs and the best-known capture site of the species in Canada, to parameterise multi-gear occupancy models for understanding the relationship between pugnose minnow occupancy and microhabitat features, including the role of turbidity. Almost 300 pugnose minnow were captured, representing the largest single collection of the species in Canadian history. The best occupancy model indicated that the probability of pugnose minnow occupancy was highest in the deepest sites with the lowest water clarity (i.e. high turbidity); however, competing models suggested that occupancy was highest at sites with wild celery (Vallisneria americana) and higher water clarity, signifying that habitats with low turbidity may be utilised if sufficient physical cover exists. Together, our results suggest that Canadian pugnose minnow populations occupy and potentially favour turbid conditions, possibly to avoid visual predators in clearer habitats. It remains uncertain whether this abiotic association represents a long-term, viable, local adaptation or whether persistence of pugnose minnow in the Canard River is at risk unless significant water quality improvements can be made.     

Stability in reproductive timing and habitat usage of Chinook salmon across six years of varying environmental conditions and abundance

The spatial and temporal distribution of spawning activity by autumn‐run Chinook salmon Oncorhynchus tshawytscha (Walbaum) was examined across multiple years. The study period included two years of extreme drought conditions when water temperatures in the spawning reach of the Stanislaus River were considered sub‐optimal for spawning and egg incubation. Despite varying levels of superimposition, redd counts and densities remained stable and positively associated with river location, indicating that superimposition may be driven by habitat preference rather than the absence of suitable spawning locations. Spawning occurred slightly later during drought years (6–10 days later compared with wetter years). This delay was attributable to deferred migration instead of deferred spawning, and the majority of redds were constructed at water temperatures exceeding the optimal temperature range. As a consequence, estimates of juvenile production during 2014 and 2015 were among the lowest on record. These findings may be related in part to the high hatchery contribution to the population, above‐average temperatures during spawning and incubation, and superimposition rates. Management recommendations include adequate cold‐water storage in the upstream reservoir, refined spawning habitat restoration techniques in the light of superimposition rates and, on a broader system scale, actions that reduce the amount of stray hatchery Chinook salmon.     

High-throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli

Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.     

First record of Ceroplastes japonicus (Gray) (Hemiptera: Coccidae) in Greece and a combined approach of morphological identification and DNA barcoding

High levels of infestation of a wax soft scale insect (Hemiptera: Coccidae) have been observed on Laurus nobilis L. (Lauraceae) and Diospyros kaki Thunb. (Ebanaceae) in Greece in recent years. Specimens of Ceroplastes were collected from regions of Thessaloniki, Naousa and Kavala (northern Greece) and Attica (southern Greece) in 2012–2017. The specimens were identified based on DNA barcoding and female morphological characteristics as Ceroplastes japonicus Green (Hemiptera: Coccidae).     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

Characterization and pathogenicity of six Cytospora strains causing stem canker of wild apple in the Tianshan Forest,China

Cytospora species are capable of causing destructive cankers of stems belonging to a wide range of woody plant species. In severe cases, cankers may lead to dieback of twigs and branches. Little is known about the Cytospora species causing canker disease of wild apple (Malus sieversii) trees in the Wild Fruit Forest Reserve in Tianshan Forest, Xinjiang Uygur Autonomous Region, China. In this study, six Cytospora isolates belonging to two species were isolated from cankerous lesions of wild apple twigs. Based on multi‐locus phylogenetic analysis using three DNA markers (ITS, tef1‐α and tub2) and morphological characterization, these isolates were identified as Cytospora mali and Cytospora parasitica. Temperature trials (15, 20, 25 and 30°C) showed that the optimal growth temperature for six isolates was 25°C. At a variety of temperatures, C. mali isolates tended to grow faster than isolates of C. parasitica, with the C. mali isolate, EGI1 performing better than others with regard to growth rate. Morphological observations showed that these species exhibited a single locule without conceptacles, and the conidia length was 3–5 μm. In vitro inoculation trials of twigs and leaves of M. sieversii seedlings revealed that the C. mali isolates were highly virulent phytopathogenic fungi, whereas the C. parasitica isolates were less virulent. The isolate EGI1 was the most virulent isolate among the six isolates. This paper presents the first report of pathogenic Cytospora spp. of the M. sieversii Tianshan Wild Fruit Forest Reserve of Yili, Xinjiang in China. It will aid in the understanding of how apple tree cankers are induced and provide disease management guidelines for M. sieversii forest conservation.     

Water availability affects nectar sugar production and insect visitation of the cup plant Silphium perfoliatum L. (Asteraceae)

The perennial cup plant (Silphium perfoliatum L.) is considered as an alternative feedstock to maize for biogas plants. Due to its ecological advantages of an extensive management and function as food resource for pollinators, it can be grown in Ecological Focus Areas (EFAs) since 2018. However, studies at the Julius Kühn-Institute in Braunschweig (Germany) showed that the assumed advantage of the cup plant of a high drought tolerance could not be confirmed regarding aspects of crop production and yield. We complemented this experiment by assessing how different soil moisture conditions affect the production of floral resources and insect visitation. In 2014, we sampled three irrigated and three rainfed plots of the cup plant. We quantified the nectar volume and sugar mass per inflorescence, the number of inflorescences per plant and calculated the total nectar sugar production. We further counted insect visitation on the inflorescences. Due to reduced numbers of inflorescences per plant and an earlier harvest, the estimated nectar sugar production was 58 kg/ha regarding irrigated and 20 kg/ha regarding rainfed plots. Honeybee visitation per inflorescence was about twice as high in the irrigated plots. Furthermore, the early harvest is a disadvantage for wild pollinators with a late activity period.     

Piscine orthoreovirus: Biology and distribution in farmed and wild fish

Piscine orthoreovirus (PRV) is a common and widely distributed virus of salmonids. Since its discovery in 2010, the virus has been detected in wild and farmed stocks from North America, South America, Europe and East Asia in both fresh and salt water environments. Phylogenetic analysis suggests three distinct genogroups of PRV with generally discrete host tropisms and/or regional patterns. PRV-1 is found mainly in Atlantic (Salmo salar), Chinook (Oncorhynchus tshawytscha) and Coho (Oncorhynchus kisutch) Salmon of Europe and the Americas; PRV-2 has only been detected in Coho Salmon of Japan; and PRV-3 has been reported primarily in Rainbow Trout (Oncorhynchus mykiss) in Europe. All three genotypes can establish high-load systemic infections by targeting red blood cells for principal replication. Each genotype has also demonstrated potential to cause circulatory disease. At the same time, high-load PRV infections occur in non-diseased salmon and trout, indicating a complexity for defining PRV's role in disease aetiology. Here, we summarize the current body of knowledge regarding PRV following 10 years of study.