Vibrio vulnificus is a potentially zoonotic bacterial pathogen of fish, which can infect humans (causing necrotic fasciitis). We analysed 24 V. vulnificus isolates (from 23 severe eel disease ...outbreaks in 8 Dutch eel farms during 1996 to 2009, and 1 clinical strain from an eel farmer) for genetic correlation and zoonotic potential. Strains were typed using biotyping and molecular typing by high-throughput multilocus sequence typing (hiMLST) and REP-PCR (Diversilab®). We identified 19 strains of biotype 1 and 5 of biotype 2 (4 from eels, 1 from the eel farmer), that were subdivided into 8 MLST types (ST) according to the international standard method. This is the first report of V. vulnificus biotype 1 outbreaks in Dutch eel farms. Seven of the 8 STs, of unknown zoonotic potential, were newly identified and were deposited in the MLST database. The REP-PCR and the MLST were highly concordant, indicating that the REP-PCR is a useful alternative for MLST. The strains isolated from the farmer and his eels were ST 112, a known potential zoonotic strain. Antimicrobial resistance to cefoxitin was found in most of the V. vulnificus strains, and an increasing resistance to quinolones, trimethoprim + sulphonamide and tetracycline was found over time in strain ST 140. Virulence testing of isolates from diseased eels is recommended, and medical practitioners should be informed about the potential risk of zoonotic infections by V. vulnificus from eels for the prevention of infection especially among high-risk individuals. Additional use of molecular typing methods such as hiMLST and Diversilab® is recommended for epidemiological purposes during V. vulnificus outbreaks.
Cyprinid herpesvirus 3 (CyHV-3) or koi herpesvirus (KHV) is a devastating virus of carp. Using generic primers for the DNA polymerase and the major capsid protein genes of cyprinid herpesviruses, ...nucleotide sequences divergent from previously described CyHV-3 were obtained. At least 3 novel groups of putative CyHV-3-like viruses were identified, sharing 95 to 98% nucleotide identity with CyHV-3 strains. Carp carrying the CyHV-3 variants did not show clinical signs consistent with CyHV-3 infection and originated from locations with no actual CyHV-3 outbreaks. These strains might represent low- or non-pathogenic variants of CyHV-3.
Summary
Background
Sets of genital swabs are routinely taken from horses to screen for the presence of Taylorella equigenitalis, the cause of contagious equine metritis. Typically, two to four ...different sites are swabbed at a time and tested by culture or PCR.
Objectives
This study explored the feasibility of pooling these swabs for a single PCR test per animal instead of testing each swab individually.
Study design
In vitro.
Methods
PCR signal strengths (Ct values) from 149 historical PCR positive genital swabs, together with historical data on the number of swabs in a set expected to be positive, were used to assess the suitability of pooling for screening horses for T. equigenitalis infection in the population at large. Twenty‐four sets of four equine genital swabs were tested. The sets were prepared in the laboratory using one or more swabs positive for T. equigenitalis from naturally infected cases. Positive and negative swabs were selected to reflect a typical range of PCR Ct values expected in field cases of T. equigenitalis infection. These pools were tested by an established PCR to assess the impact and suitability of a PCR test on pooled swabs compared to individual swab testing, by comparing the Ct values.
Results
Pooling one positive swab with three negative swabs produced a small drop in Ct value but all pools were still clearly positive.
Main limitations
Large numbers of field positive horses are not available, but the proof of concept approach with laboratory prepared pools shows the method is applicable to field cases.
Conclusions
It was concluded that pooling of swabs would confer no appreciable drop in the ability to detect a positive animal compared to individual swab testing; pooling is therefore a suitable alternative to individual swab testing with reduced costs.
The Summary is available in Spanish – see Supporting Information
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•We show for the first time presence of Aphanomyces astaci in the Netherlands.•Four out of five tested North American alien crayfish taxa were found infected.•Out of two newly studied ...taxa, Orconectes virilis was shown to be an A. astaci host.•Its carrier status was independently confirmed in Dutch and English populations.•The infection was also detected in Dutch Chinese mitten crabs (Eriocheir sinesis).
North American crayfish species as hosts for the crayfish plague pathogen Aphanomyces astaci contribute to the decline of native European crayfish populations. At least six American crayfish species have been reported in the Netherlands but the presence of this pathogenic oomycete with substantial conservational impact has not yet been confirmed in the country. We evaluated A. astaci prevalence in Dutch populations of six alien crustaceans using species-specific quantitative PCR. These included three confirmed crayfish carriers (Orconectes limosus, Pacifastacus leniusculus, Procambarus clarkii), two recently introduced but yet unstudied crayfish (Orconectes cf. virilis, Procambarus cf. acutus), and a catadromous crab Eriocheir sinensis. Moderate levels of infection were observed in some populations of O. limosus and P. leniusculus. Positive results were also obtained for E. sinensis and two Dutch populations of O. cf. virilis. English population of the latter species was also found infected, confirming this taxon as another A. astaci carrier in European waters. In contrast, Dutch P. clarkii seem only sporadically infected, and the pathogen was not yet detected in P. cf. acutus. Our study is the first confirmation of crayfish plague infections in the Netherlands and demonstrates substantial variation in A. astaci prevalence among potential hosts within a single region, a pattern possibly linked to their introduction history and coexistence.
The Wadden Sea is an extensive wetland area, recognized as UNESCO world heritage site of international importance. Since the mid‐1990s, the invasive Pacific oyster Crassostrea gigas (Thunberg 1793) ...population in the area has grown exponentially, having a distinct impact on the ecosystem. The recent spread of the emerging oyster pathogen Ostreid herpesvirus OsHV‐1 μVar worldwide and specifically in the oyster culture areas in the south of the Netherlands raised the question whether the virus may also be present in the Wadden Sea. In the summer of 2012 juvenile Pacific oysters were collected from five locations in the Dutch Wadden Sea. The virus was shown to be present in three of the five locations by real‐time PCR and sequencing. It was concluded that OsHV‐1 μVar has settled itself in Pacific oyster reefs in the Wadden Sea. These results and the recent discoveries of OsHV‐1 microvariants in Australia and Korea indicate that OsHV‐1 μVar and related variants might be more widespread than can be deduced from current literature. In particular in regions with no commercial oyster culture, similar to the Wadden Sea, the virus may go undetected as wild beds with mixed age classes hamper the detection of mortality among juvenile oysters.
The worldwide decline of the eel population is thought to be caused by several factors, among which eel diseases. To investigate diseases of European silver eels
Anguilla anguilla in the Netherlands, ...in Nov–Dec 2004 12 silver eels, and in Aug–Dec 2005 80 eels were caught in downstream parts (rivers) of the River Rhine and in Lake IJsselmeer. The eels were measured and weighed, necrotized, and individually checked for presence of external and internal parasites, bacterial infections and viruses, and blood smears were made for haematology. This is the first multidisciplinary study of the health of Dutch silver eels. In the small (2
×
6 eels) pilot study of 2004 in the River Rhine and Lake IJsselmeer respectively, most eels showed aspecific fin haemorrhages, some had ectoparasites, nearly none had parasites in the intestine, half of the groups had
Anguillicola crassus in their swimbladder, only few had
Trypanosoma's in their blood, and no primary virus or bacterial infections were found, although one eel from Lake IJsselmeer was positive for anguillid herpesvirus 1 (AngHV-1, former Herpesvirus anguillae, HVA) in the PCR test only. The blood of all 6 and 2/6 of the eels respectively was considered abnormal, and the eels had a proper condition.
In 2005, in 50 eels from the River Rhine and 30 from Lake IJsselmeer respectively, again aspecific fin haemorrhages were often seen, some of the eels had ectoparasites and parasites in the intestine, most eels had
A. crassus in their swimbladder, 32% and 53% had
Trypanosoma's in their blood, from 44% and 13% of the eels AngHV-1 was isolated, and 44% and 27% were tested positive by PCR, with a peak in August, 10% of both groups of eels had an internal bacterial infection, mostly due to
Aeromonas spp. The blood of about half of the eels was considered abnormal, but the eels had a proper condition.
It was concluded, that the silver eels of this study had a proper Fulton condition factor (values 2.00–2.26), with aspecific fin haemorrhages, often were
Trypanosoma-,
A. crassus- and AngHV-1-infected, dependent on the season, and often showed an abnormal haematology.
A. crassus causes injuries and is a chronic stress factor, more than the other parasites, which were mostly found less prevalent. In fact, lymphocytosis was directly related to
A. crassus infection. Stress and injuries by
A. crassus might induce disease through the presence of virus (AngHV-1), relevant in the health status of the silver eels during their spawning migration if ambient water temperatures would enhance a clinical infection of AngHV-1 disease. Moreover this virus might potentially decrease the survival of the silver eels by itself, because spawning migration to the Sargasso Sea takes wild eels to temperate/tropical areas in which the clinical infection by AngHV-1 is surely enhanced.
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► A Flat oyster health survey was carried out in the Spanish Mediterranean coast. ► Marteilia sp. and Bonamia sp. parasites were detected by different techniques. ► SYBR Green ...real-time PCR allowed efficiently Bonamia detection. ► Phylogenetical studies corroborate the presence of Bonamia exitiosa. ► This is the first detection of B. exitiosa in the Spanish Mediterranean.
Bonamia exitiosa and Bonamia ostreae are parasites that reproduce within the haemocytes of several oyster species. In Europe, the host species is the flat oyster Ostrea edulis. The parasite B. ostreae has been responsible for mortalities since the late 1970s throughout the European Atlantic coast. B. exitiosa was first detected, in 2007, on this continent in flat oysters cultured in Galicia (NW Spain). Since then, the parasite has also been detected in France, Italy and the United Kingdom. The bays of the Ebro Delta in the south of Catalonia represent the main bivalve culture area in the Mediterranean coast of Spain. Previous information from the area includes reports of several flat oyster pathogens, including the notifiable parasite Marteilia refringens. However, the status with regard to Bonamia parasites was uncertain. In the present study, a Bonamia parasite was observed in flat oysters cultured in the Alfacs Bay of the Ebro Delta by histology and real-time PCR. PCR-RFLP and sequencing suggested the presence of B. exitiosa. Finally, phylogenetic analyses of the studied Bonamia isolates corroborated B. exitiosa infection. M. refringens was also observed in the same oyster batch, and co-infection with both parasites was also detected. This is the first detection of B. exitiosa, in Catalonia and the Spanish Mediterranean coast. The impact of the parasite on the Mediterranean flat oyster activity needs to be urgently addressed.
Organisms of the genus Bonamia are intracellular protistan parasites of oysters. To date, 4 species have been described (B. ostreae, B. exitiosa, B. perspora and B. roughleyi), although the status of ...B. roughleyi is controversial. Introduction especially of B. ostreae and B. exitiosa to naïve host populations has been shown to cause mass mortalities in the past and has had a dramatic impact on oyster production. Both B. ostreae and B. exitiosa are pathogens notifiable to the World Organisation for Animal Health (OIE) and the European Union. Effective management of the disease caused by these pathogens is complicated by the extensive nature of the oyster production process and limited options for disease control of the cultured stocks in open water. This review focuses on the recent advances in research on genetic relationships between Bonamia isolates, geographical distribution, susceptible host species, diagnostics, epizootiology, host-parasite interactions, and disease resistance and control of this globally important genus of oyster pathogens.
The ultrastructure of Bonamia from Ostrea angasi from Australia, Crassostrea ariakensis from the USA, O. puelchana from Argentina and O. edulis from Spain was compared with described Bonamia spp. All ...appear conspecific with B. exitiosa. The Bonamia sp. from Chile had similarities to the type B. exitiosa from New Zealand (NZ), but less so than the other forms recognized as B. exitiosa. Two groups of ultrastructural features were identified; those associated with metabolism (mitochondrial profiles, lipid droplets and endoplasmic reticulum), and those associated with haplosporogenesis (Golgi, indentations in the nuclear surface, the putative trans-Golgi network, perinuclear granular material and haplosporosome-like bodies). Metabolic features were regarded as having little taxonomic value, and as the process of haplosporogenesis is not understood, only haplosporosome shape and size may be of taxonomic value. However, the uni-nucleate stages of spore-forming haplosporidians are poorly known and may be confused with Bonamia spp. uni-nucleate stages. The many forms of NZ B. exitiosa have not been observed in other hosts, which may indicate that it has a plastic life cycle. Although there are similarities between NZ B. exitiosa and Chilean Bonamia in the development of a larger uni-nucleate stage and the occurrence of cylindrical confronting cisternae, the clarification of the identity of Chilean Bonamia must await molecular studies.
The Baltic tellin Limecola balthica is one of the most common bivalves in intertidal areas in the Northern Hemisphere. Over the last 2 decades, the species has been suffering from a decrease in adult ...survival in the European Wadden Sea. While several factors such as global warming and fisheries have been suggested to influence the population dynamics of this bivalve mollusc, the potential role of diseases has never been investigated. In this study, we investigated whether disseminated neoplasia, a common proliferative disorder in bivalve molluscs, could play a potential role in the recent population decline of Baltic tellins in the Wadden Sea. We conducted a field survey in the Dutch Wadden Sea to (1) determine whether the disease occurs in Baltic tellins in the Wadden Sea and (2) quantify the occurrence and severity of the disease via histology. Disseminated neoplasia occurred in L. balthica at each of the 10 sampled locations with very high prevalences (21-89%) compared to those reported elsewhere for this species. The highest severity category was found in 8 to 87% of affected individuals, with severity generally increasing with prevalence. Disseminated neoplasia usually increases mortality among affected individuals and may also be associated with important sub-lethal effects, especially regarding gametogenesis. Thus, we suggest that disseminated neoplasia may play a key role in the population dynamics of the Baltic tellin, the extent of which remains to be investigated in future studies.
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