Virus diseases in aquaculture

World Journal of Microbiology and Biotechnology -     

The asexual pre-cyst development of Sarcocystis tenella in experimentally infected specific pathogen-free lambs

Twenty-two lambs raised under sporozoa-free conditions were infected at weaning with 0.25–2.0 million Sarcocystis tenella sporocysts obtained from dogs and then necropsied at intervals between 1.3 h and 41 days post-inoculation (dpi). Fixed and frozen sections from 47 different tissues from each lamb were examined respectively by a variety of histochemical stains and by indirect fluorescent-antibody staining. The remnants of excysted sporocysts were found within the gastro-intestinal tract between 1.3 h and 3 dpi. No enteric proliferative stage of the parasite was detected. First-generation meronts (schizonts) were detected from 6 to 19 dpi within the endothelial cells of arterioles in most organs and tissues. The meronts were undifferentiated from 6 to 9 dpi but were mature in appearance from 12 to 19 dpi (measuring 22.3 × 13.4 μm and containing 18–28 merozoites). Second-generation meronts were detected from 21 to 34 dpi within the endothelial cells of capillaries throughout the entire body. They were undifferentiated from 21 to 23 dpi but appeared mature from 25 to 34 dpi (measuring 15.2 × 10.6 μm and containing 18–38 merozoites). Several smaller meronts were then detected at 36 dpi within cells of the mononuclear phagocytic system. They were all mature in appearance (measuring 7.4 × 5.1 μm and containing 6–9 merozoites) and are thought to have formed facultatively following the incorporation of some second-generation merozoites into phagocytic cells. Only developing cysts were then detected at 41 dpi throughout the cardiac and skeletal musculature. Elements in both first- and second-generation merozoites stained markedly with haematoxylin and eosin, periodic acid-Schiff's, alkaline Giemsa-colophonium and basic fuchsin stains. However, only second-generation meronts and developing cysts exhibited strong specific reactions with the fluorescent-antibody stain.     

Virulence of luminescent and non-luminescent isogenic vibrios towards gnotobiotic Artemia franciscana larvae and specific pathogen-free Litopenaeus vannamei shrimp

Aims:  This study was conducted to test the virulence of luminescent (L) and non-luminescent (NL) isogenic strains of Vibrio campbellii LMG21363, Vibrio harveyi BB120 (wild type) and quorum-sensing mutant strains derived from the wild type such as Vibrio harveyi BB152, BB170, MM30 and BB886.
Methods and Results:  The NL strains could be obtained by culturing rifampicin-resistant luminescent strains in the dark under static condition. The virulence of the L and NL strains was tested in gnotobiotic Artemia franciscana larvae challenged with 10⁴ CFU ml⁻¹ of bacteria. All luminescent isogenic tested strains showed higher virulence compared to the NL strains. The virulence of L and NL V. campbellii and V. harveyi BB120 was also tested in specific pathogen-free juvenile shrimp upon intramuscular injection with 10⁶ CFU of bacteria. In contrast with Artemia , there was no significant difference in mortality between the groups challenged with L and NL strains ( P  > 0·05). The non-luminescent strains were not able to revert back to the luminescent state and quorum sensing did not influence this phenotypic shift.
Conclusions:  Luminescent Vibrio strains can switch to a non-luminescent state by culturing them in static conditions. The NL strains become less virulent as verified in Artemia .
Significance and Impact of the Study:  The luminescent state of Vibrio cells in a culture needs to be verified in order to assure maintenance of virulence.     

Taura syndrome,a disease important to shrimp farms in the Americas

World Journal of Microbiology and Biotechnology -     

Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): a review

Penaeid shrimp aquaculture is an important industry in the Americas, and the industry is based almost entirely on the culture of the Pacific White Shrimp, Litopenaeus vannamei. Western Hemisphere shrimp farmers in 14 countries in 2004 produced more than 200,000 metric tons of shrimp, generated more than $2 billion in revenue, and employed more than 500,000 people. Disease has had a major impact on shrimp aquaculture in the Americas since it became a significant commercial entity in the 1970s. Diseases due to viruses, rickettsial-like bacteria, true bacteria, protozoa, and fungi have emerged as major diseases of farmed shrimp in the region. Many of the bacterial, fungal and protozoan caused diseases are managed using improved culture practices, routine sanitation, and the use of chemotherapeutics. However, the virus diseases have been far more problematic to manage and they have been responsible for the most costly epizootics. Examples include the Taura syndrome pandemic that began in 1991-1992 when the disease emerged in Ecuador, and the subsequent White Spot Disease pandemic that followed its introduction to Central America from Asia in 1999. Because of their socioeconomic significance to shrimp farming, seven of the nine crustacean diseases listed by the World Animal Organization (OIE) are virus diseases of shrimp. Of the seven virus diseases of penaeid shrimp, five are native to the Americas or have become enzootic following their introduction. The shrimp virus diseases in the Americas are increasingly being managed by exclusion using a combination of biosecurity and the practice of culturing domesticated specific pathogen-free (SPF) stocks or specific pathogen-resistant (SPR) stocks. Despite the significant challenges posed by disease, the shrimp farming industry of the Americas has responded to the challenges posed by disease and it has developed methods to manage its diseases and mature into a sustainable industry.     

The use of probiotics in shrimp aquaculture

Shrimp aquaculture, as well as other industries, constantly requires new techniques in order to increase production yield. Modern technologies and other sciences such as biotechnology and microbiology are important tools that could lead to a higher quality and greater quantity of products. Feeding and new practices in farming usually play an important role in aquaculture, and the addition of various additives to a balanced feed formula to achieve better growth is a common practice of many fish and shrimp feed manufacturers and farmers. Probiotics, as 'bio-friendly agents' such as lactic acid bacteria and Bacillus spp., can be introduced into the culture environment to control and compete with pathogenic bacteria as well as to promote the growth of the cultured organisms. In addition, probiotics are nonpathogenic and nontoxic microorganisms without undesirable side-effects when administered to aquatic organisms. These strains of bacteria have many other positive effects, which are described in this article.     

The role of selective breeding and biosecurity in the prevention of disease in penaeid shrimp aquaculture

About 3.5 million metric tons of farmed shrimp were produced globally in 2009 with an estimated value greater than USD$14.6 billion. Despite the economic importance of farmed shrimp, the global shrimp farming industry continues to be plagued by disease. There are a number of strategies a shrimp farmer can employ to mitigate crop loss from disease, including the use of Specific Pathogen Free (SPF), selectively bred shrimp and the adoption of on-farm biosecurity practices. Selective breeding for disease resistance began in the mid 1990s in response to outbreaks of Taura syndrome, caused by Taura syndrome virus (TSV), which devastated populations of farmed shrimp (Litopenaeus vannamei) throughout the Americas. Breeding programs designed to enhance TSV survival have generated valuable information about the quantitative genetics of disease resistance in shrimp and have produced shrimp families which exhibit high survival after TSV exposure. The commercial availability of these selected shrimp has benefitted the shrimp farming industry and TSV is no longer considered a major threat in many shrimp farming regions. Although selective breeding has been valuable in combating TSV, this approach has not been effective for other viral pathogens and selective breeding may not be the most effective strategy for the long-term viability of the industry. Cost-effective, on-farm biosecurity protocols can be more practical and less expensive than breeding programs designed to enhance disease resistance. Of particular importance is the use of SPF shrimp stocked in biosecure environments where physical barriers are in place to mitigate the introduction and spread of virulent pathogens.     

新工科背景下高校生物安全人才培养体系建设北大核心CSCD

近年来,全球生物安全形势日趋严峻,重大传染病疫情、生物技术误用和谬用、生物武器等生物安全威胁不断显现,生物安全已跃升为我国国家安全的重要组成部分;而与此同时,生物安全作为一门新兴的交叉型学科,其教育体系的建立与完善还处于起步阶段。受国内外生物安全形势、生物安全产业发展以及国家安全学学科设立的影响,凸显了我国在生物安全这一交叉领域的人才储备的短板和不足。尤其在新工科建设背景下,更加显现出生物安全人才培养的重要性。本文根据作者在实际教学中的经验,从建立交叉融合课程体系、构建创新性教学模式、探索通识教育改革方案和完善持续改进机制等几个方面,系统介绍了新工科背景下完善生物安全领域人才体系的具体措施。     

Probiotics in aquaculture: importance and future perspectives

Aquaculture is one of the fastest developing growth sectors in the world and Asia presently contributes about 90% to the global production. However, disease outbreaks are constraint to aquaculture production thereby affects both economic development of the country and socio-economic status of the local people in many countries of Asia-Pacifi c region. Disease control in aquaculture industry has been achieved by following different methods using traditional ways, synthetic chemicals and antibiotics. However, the use of such expensive chemotherapeutants for controlling diseases has been widely criticized for their negative impacts like accumulation of residues, development of drug resistance, immunosuppressants and reduced consumer preference for aqua products treated with antibiotics and traditional methods are ineffective against controlling new diseases in large aquaculture systems. Therefore, alternative methods need to be developed to maintain a healthy microbial environment in the aquaculture systems there by to maintain the health of the cultured organisms. Use of probiotics is one of such method that is gaining importance in controlling potential pathogens. This review provides a summary of the criteria for the selection of the potential probiotics, their importance and future perspectives in aquaculture industry.     

Herpes B-virus seroreactivity in a colony of Macaca mulatta: data from the Sabana Seca Field Station, a new specific pathogen-free program

The demand for B-virus-free animals for biomedical research is increasing, while at the same time the availability of such animals is decreasing. The establishment of Specific Pathogen-Free (SPF) breeding macaque colonies is a priority of the National Institutes of Health. Nevertheless, it is well known that seroreactivity to B-virus can be difficult to interpret, particularly as it can vary over time in a single animal. The aim of the present study was to implement a reliable algorithm to examine B-virus reactivity among the rhesus monkey population of the Caribbean Primate Research Center. The sensitivity and specificity of our assay were determined using reports from two different laboratories as references. Whereas 95.4% of animals showed consistent serological status and 4.6% of animals recruited to this SPF program showed serovariability to B-virus over the initial 2 years of examination. Implications for all SPF programs are discussed in this article.     

Cultivar‐specific effects of pathogen testing on storage root yield of sweetpotato,Ipomoea batatas

The accumulation and perpetuation of viral pathogens over generations of clonal propagation in crop species such as sweetpotato, Ipomoea batatas, inevitably result in a reduction in crop yield and quality. This study was conducted at Bundaberg, Australia to compare the productivity of field‐derived and pathogen‐tested (PT) clones of 14 sweetpotato cultivars and the yield benefits of using healthy planting materials. The field‐derived clonal materials were exposed to the endemic viruses, while the PT clones were subjected to thermotherapy and meristem‐tip culture to eliminate viral pathogens. The plants were indexed for viruses using nitrocellulose membrane‐enzyme‐linked immunosorbent assay and graft‐inoculations onto Ipomoea setosa. A net benefit of 38% in storage root yield was realised from using PT materials in this study. Conversely, in a similar study previously conducted at Kerevat, Papua New Guinea (PNG), a net deficit of 36% was realised. This reinforced our finding that the response to pathogen testing was cultivar dependent and that the PNG cultivars in these studies generally exhibited increased tolerance to the endemic viruses present at the respective trial sites as manifested in their lack of response from the use of PT clones. They may be useful sources for future resistance breeding efforts. Nonetheless, the potential economic gain from using PT stocks necessitates the use of pathogen testing on virus‐susceptible commercial cultivars.     

Eggs of the Australian saltmarsh mosquito,Aedes camptorhynchus,survive for long periods and hatch in instalments: implications for biosecurity in New Zealand

The Australian saltmarsh mosquito, Aedes camptorhynchus (Diptera: Culicidae), is a significant biting pest and disease vector and is the subject of an eradication programme in New Zealand (NZ), where it has been resident for more than 10 years. To better understand the ecology of this common and widespread pest, we studied egg longevity and hatching patterns in the laboratory. By regularly testing for the presence of viable embryos, we found that eggs may last more than 15 months when stored dry (13% viable at this time). Eggs display instalment hatching, with no more than 56% of a batch hatching upon first inundation. Further hatching may occur for at least six inundations and some unhatched eggs may remain viable even after this. Variation in hatching rates can be observed using different water types, with weaker hatching media stimulating lower hatching rates spread over more inundations. By applying average hatching rates to a non‐linear model of natural egg attrition, we showed that egg batches exposed to three inundations should be exhausted (zero live eggs present) in approximately 11 months at the conditions tested here. These findings have implications for the current eradication programme for Ae. camptorhynchus in NZ and for our understanding of the ecology of a widespread and common disease vector in Australia.     

Planktonic bacteria in white shrimp (Litopenaeus vannamei) and channel catfish (Letalurus punetaus) aquaculture ponds in a salt-alkaline region

Aquaculture in salt-alkaline regions is encouraged in China, and culture of many aquatic species has been introduced into these areas. In this study, we cultured two species, white shrimp (Litopenaeus vannamei) and channel catfish (Letalurus punetaus) separately in aquaculture ponds in a salt-alkaline region in northwest China and assessed the impacts of the aquaculture operations on the planktonic bacterial community in the culture ponds. Culture of both species decreased the planktonic bacterial diversity and altered the bacterial community structure in the aquaculture ponds compared with the source water. Among the 10 dominant bacterial phyla, 8 were significantly correlated with environmental parameters; the exception was Actinobacteriota, the most dominant phylum, and Firmicutes. Proteobacteria and Bacteroidota abundances showed significant positive correlations with alkalinity, whereas Patescibacteria, Cyanobacteria, Planctomycetota, and Verrucomicrobiota abundance were positively correlated with salinity. Linear regression analysis showed that alkalinity was positively correlated with bacterial beta diversity and salinity was negatively correlated with that. In addition, white shrimp aquaculture significantly lowered the alkalinity, which suggests that culture of this species in inland salt-alkaline regions is a potential dealkalization solution.     

Individual discrimination in the big-clawed snapping shrimp, Alpheus heterochelis

Rahman et al. (Rahman, N., Dunham, D.W. and Govind, C.K. (2001). Mate recognition and pairing in the big-clawed snapping shrimp, Alpheus herterochelis. Mar. Fresh. Behav. Physiol., 34, 213-226.) demonstrated discrimination by snapping shrimp between former mates and unfamiliar conspecifics, but did not test individual discrimination. In the present study, snapping shrimp showed discrimination between familiar and unfamiliar same-sex conspecifics by preferentially entering that arm of a Y-maze leading to familiar individuals. Furthermore, after being exposed to water from the home tanks of unknown individuals, they later showed an elevated response to this water, if the direction from which the water came into their tank was changed to be novel. This indicates that test subjects associated a familiar chemical stimulus with its location in the environment. This discrimination could only have been made if that chemical signature were recognised as different from that of another chemically familiar individual. This result also demonstrates that the water surrounding an individual contains sufficient (chemical) information to allow discrimination of one individual from another.     

Individual discrimination in the big-clawed snapping shrimp,Alpheus heterochelis

Rahman et al. (Rahman, N., Dunham, D.W. and Govind, C.K. (). Mate recognition and pairing in the big-clawed snapping shrimp, Alpheus herterochelis. Mar. Fresh. Behav. Physiol., 34, 213–226.) demonstrated discrimination by snapping shrimp between former mates and unfamiliar conspecifics, but did not test individual discrimination. In the present study, snapping shrimp showed discrimination between familiar and unfamiliar same-sex conspecifics by preferentially entering that arm of a Y-maze leading to familiar individuals. Furthermore, after being exposed to water from the home tanks of unknown individuals, they later showed an elevated response to this water, if the direction from which the water came into their tank was changed to be novel. This indicates that test subjects associated a familiar chemical stimulus with its location in the environment. This discrimination could only have been made if that chemical signature were recognised as different from that of another chemically familiar individual. This result also demonstrates that the water surrounding an individual contains sufficient (chemical) information to allow discrimination of one individual from another.     

Ex situ transportation of coral larvae for research,conservation, and aquaculture

Due to the lack of appropriate methods to transport high amounts of larvae ex situ over large distances, the availability of coral larvae was so far mainly limited to their place of origin. For a research project at Rotterdam Zoo, The Netherlands, we transported several thousand larvae of three broadcast spawners (Acropora tenuis, A. digitifera, Diploria strigosa) from the Indo Pacific and the Caribbean to Europe. Beside logistics and packing techniques, post-transport survival rates were mainly influenced by larvae density and transport duration. Our results indicate optimum survival rates of >90% at densities of 4 larvae ml⁻¹ when not exceeding a transportation time of 4 days. The ex situ transport of coral larvae over large distances might offer new possibilities for research, conservation, and aquaculture.     

The biosecurity status and its associations with production and management characteristics in farrow-to-finish pig herds

Disease prevention through biosecurity measures is believed to be an important factor for improvement of the overall health status in animal production. This study aimed at assessing the levels of implementation of biosecurity measures in pig production in four European Union (EU) countries and to describe possible associations between the biosecurity level and farm and production characteristics. A cross-sectional study was conducted in 232 farrow-to-finish pig herds in Belgium, France, Germany and Sweden between December 2012 and December 2013. The biosecurity status in each of these herds was described and quantified by using the risk-based scoring tool Biocheck.UGent^TM (www.biocheck.ugent.be). Production and management characteristics, obtained from the herd management system and by interviewing the farmer, were analysed for their association with the biosecurity level. A causal path was designed to study statistical associations. The results showed that there was substantial room for improvement in the biosecurity status on many pig farms. Significant differences (P<0.01) both in internal and external biosecurity levels were observed between countries. The external biosecurity status, combining all measures taken to prevent disease introduction into the herd, was highest in Germany and lowest in France. The internal biosecurity status, combining all measures taken to prevent within herd disease transmission, was highest in Sweden and lowest, with a large variation, in Belgium. External biosecurity scores were in general higher compared to internal biosecurity scores. The number of pathogens vaccinated against was significantly associated with internal biosecurity status, suggesting an overall more preventive approach towards the risk of disease transmission. A higher external biosecurity was associated with more weaned piglets per sow per year. Furthermore also the weaning age and the mortality till weaning were highly associated with the number of weaned piglets per sow per year. The negative association observed between the biosecurity level and the estimated frequency of treatment against certain clinical signs of disease as a proxy for disease incidence is consistent with the hypothesis that a higher biosecurity level results in healthier animals. These findings promote an improved biosecurity status at pig farms and are of relevance in the discussion on alternative ways to keep animals healthy with a reduced necessity of antimicrobials; Prevention is better than cure!     

Histopathology of aflatoxicosis in the marine shrimp Penaeus stylirostris and P. vannamei

The acute and subacute toxicity of aflatoxin B₁ to the marine shrimp Penaeus stylirostris and P. vannamei (Order: Decapoda, Class: Crustacea) was investigated. Experimental shrimp were exposed to a range of concentrations of the toxin directly by intramascular injection (from 2 to 160 μg aflatoxin B₁/g body weight), or by multiple per os dosing with the feed (from 53 to 300 μg aflatoxin B₁/g feed) for up to 25 days. The histopathogenesis of aflatoxicosis in the aflatoxin-exposed animals was followed and found to be time and dose dependent in the hepatopancreas, mandibular organ, and in the hematopoietic organs. Less significant and/or inconsistent lesions were also observed in other organs and tissues, but a time-dose dependency was not noted. The principal lesions of aflatoxicosis in penaeid shrimp occur in the hepatopancreas and the mandibular organ. In the former organ, subacute and acute aflatoxicosis is expressed as necrosis of the hepatopancreatic tubule epithelium that proceeds from the proximal (older) portion of the tubules, in the center of the organ, to the peripheral (younger) tubule tips. A marked intertubular hemocytic inflammation followed by encapsulation and fibrosis of affected tubules follows in subacute aflatoxicosis, but is not as developed as in acute aflatoxicosis. The mandibular organ in aflatoxicosis displays a necrosis of the peripheral epithelial cells of the cords within the gland that progresses proximally to the central vein. Only a slight hemocytic inflammation accompanies the degenerative changes in this latter organ.