Role of basidiomycete fungi in the grapevine trunk disease esca

Phaeomoniella chlamydospora and wood-rotting basidiomycete fungi, namely Fomitiporia spp., are known esca pathogens. However, the effect of their mixed infections and the sequence of infection on disease development is unclear. To determine the effects of single and co-inoculations on symptoms, potted Vitis vinifera ‘Crimson Seedless’ was inoculated with P. chlamydospora either alone or in combination with one of four basidiomycetes: Coprinellus radians, Fomitiporia langloisii, F. polymorpha and the novel species Tropicoporus texanus. Basidiomycetes were isolated from vines with foliar symptoms of esca in California and Texas. In sequential co-inoculations, the effects of different sequences of infection (P. chlamydospora first, basidiomycete 6 months later; and vice versa) were tested, compared to simultaneous co-inoculations. Plants inoculated with P. chlamydospora either alone or in combination with a basidiomycete (in any sequence) did not differ significantly in the length of black-streaking lesions. In plants inoculated only with a basidiomycete, the appearance of large brown lesions, coupled with the absence of this wood symptom from control plants, suggests that C. radians, F. langloisii and T. texanus are pathogenic. Foliar symptoms resembling those of esca in the field (marginal and/or interveinal scorching, combined with red and/or yellow discoloration) were statistically more frequent among plants inoculated with F. polymorpha or T. texanus, either simultaneously or following P. chlamydospora, compared to single inoculations. Sequential co-inoculations of a basidiomycete before or after P. chlamydospora were associated with similar lesion lengths, suggesting that basidiomycetes may not require infection by P. chlamydospora in order to extensively colonize the wood.     

Ultrastructural analysis of sequential cyprinid herpesvirus 3 morphogenesis in vitro

Cyprinid herpesvirus 3 (CyHV‐3) is an alloherpesvirus, and it is the aetiological agent of koi herpesvirus disease. Although the complex morphogenic stages of the replication cycle of CyHV‐3 were shown to resemble that of other members of the Herpesvirales, detailed analysis of the sequence and timing of these events was not definitively determined. This study describes these features through a time course using cyprinid cell cultures (KF‐1 and CCB) infected with CyHV‐3 (KHV isolate, H361) and analysed by transmission electron microscopy. Rapid viral entry was noted, with high levels of intracellular virus within 1–4 h post‐infection (hpi). Intranuclear capsid assembly, paracrystalline array formation and primary envelopment of capsids occurred within 4 hpi. Between 1 and 3 days post‐infection (dpi), intracytoplasmic secondary envelopment occurred, as well as budding of infectious virions at the plasma membrane. At 5–7 dpi, the cytoplasm contained cytopathic vacuoles, enveloped virions within vesicles, and abundant non‐enveloped capsids; also there was frequent nuclear deformation. Several morphological features are suggestive of inefficient viral assembly, with production of non‐infectious particles, particularly in KF‐1 cells. The timing of this alloherpesvirus morphogenesis is similar to other members of the Herpesvirales, but there may be possible implications of using different cell lines for CyHV‐3 propagation.     

High fluid shear strain causes injury in silver shark: Preliminary implications for Mekong hydropower turbine design

The Lower Mekong Basin is facing unprecedented threats to fish diversity from hydropower development. There is increasing pressure on developers and construction authorities to design solutions to improve fish survival through turbines, thus protecting the resources in regions being developed for hydropower. A hydraulic characteristic of hydropower turbines with known fisheries impacts is fluid shear stress. Elevated shear stress occurs where rapidly flowing water passes near spillways, screens and within turbine draft tubes. Shear stress can have adverse impacts on fish, but no work has assessed whether this holds true for Mekong River species. A flume was used to determine critical tolerances of silver shark, Balantiocheilos melanopterus (Bleeker), to shear stress rates at a high‐velocity jet which simulated a hydropower turbine. Fish were assessed for injury or mortality following exposure. Results were compared against a no‐shear control. Injury and mortality were greater at higher shear stress exposures. Injuries occurred at all shear exposure levels with mortality at shear levels higher than 600/s. This approach should help design future hydropower turbines if data on other species demonstrate similar results. If the likelihood of adverse impact is high, then shear stress will need to be considered in the design of future hydropower facilities.     

Cyprinid herpesvirus 3 as a potential biological control agent for carp (Cyprinus carpio) in Australia: susceptibility of non‐target species

Carp (Cyprinus carpio L.) is a pest species in Australian waterways, and cyprinid herpesvirus 3 (CyHV‐3) is being considered as a potential biological control (biocontrol) agent. An important consideration for any such agent is its target specificity. In this study, the susceptibility to CyHV‐3 of a range of non‐target species (NTS) was tested. The NTS were as follows: 13 native Australian, and one introduced, fish species; a lamprey species; a crustacean; two native amphibian species (tadpole and mature stages); two native reptilian species; chickens; and laboratory mice. Animals were exposed to 100–1000 times the approximate minimum amount of CyHV‐3 required to cause disease in carp by intraperitoneal and/or bath challenge, and then examined clinically each day over the course of 28 days post‐challenge. There were no clinical signs, mortalities or histological evidence consistent with a viral infection in a wide taxonomic range of NTS. Furthermore, there was no molecular evidence of infection with CyHV‐3, and, in particular, all RT‐PCRs for viral mRNA were negative. As a consequence, the results encourage further investigation of CyHV‐3 as a potential biocontrol agent that is specific for carp.     

Survival and behavioural responses of cool and warm water fish sedated with AQUI‐S®20E (10% eugenol) at high loading densities

This study evaluated the effects of AQUI‐S^®20E (10% eugenol) sedation on the survival and behaviour of yellow perch Perca flavescens (Mitchill) and Nile tilapia Oreochromis niloticus L. held in high loading densities. Fish were held in 0–300 mg L^?1 AQUI‐S^®20E (0–30 mg L^?1 eugenol) for up to 10 h in static tanks. At 17°C, yellow perch held in 200 and 300 mg L^?1 AQUI‐S^®20E were lightly sedated for up to 7 h. Yellow perch at 200 and 300 mg L^?1 AQUI‐S^®20E also had >95% mean survival 7‐days post exposure using loading densities up to 360 g L^?1. Nile tilapia were only sedated for ≤3 h in concentrations up to 300 mg L^?1 at 22°C and had >90% mean survival at loading densities ≤480 g L^?1. Ammonia in tanks increased significantly as loading density increased, but treatment with AQUI–S^®20E did not reduce ammonia accumulation. Results suggest that AQUI–S^®20E was effective to sedate yellow perch and Nile tilapia at high loading densities, but sedation varied with loading density and species.     

Effect of fish meal replacement with Carinata Brassica carinata in low animal protein diets of rainbow trout Oncorhynchus mykiss (Walbaum) on trypsin activity,protein and amino acid digestibility and bioavailability

Replacement of >25% of fish meal (FM) with aerobically converted carinata meal (ACCM) in low (200g/kg) animal protein (reference = 200g/kg FM) diets of rainbow trout Oncorhynchus mykiss resulted in reduced growth, partly due to reduced feed consumption and protein utilization. In this study, we determined the effect of FM replacement with ACCM on trypsin activity, apparent digestibility coefficients (ADCs) of protein, and essential amino acid (EAA) or conditionally essential amino acid (CEAA) ADCs and bioavailability. Replacement of FM did not alter protein ADCs or trypsin activity. Replacement of >25% FM reduced all EAA and CEAA ADCs except for arginine and leucine which were only reduced by 75% FM replacement. Only serum free lysine and muscle free histidine were reduced by >25% FM replacement. Muscle free lysine was only reduced by 75% FM replacement. Replacement of FM reduced EAA peak concentrations and resulted in slower release of EAAs in serum. Cumulative total EAAs in serum and muscle decreased with FM replacement. Ratios of EAAs to lysine showed that tryptophan was the most limiting EAA. However, isoleucine, leucine, methionine and phenylalanine were also inadequate for muscle synthesis for the first 9–12 hr following force‐feeding. Optimal time for protein synthesis was ≥36 hr. Although any level of FM replacement did not reduce protein ADCs and trypsin activity, replacement of ≥25% FM reduced EAA ADCs and bioavailability of lysine and histidine, which partly contributed to the observed differences in growth.     

Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.