Hyperthermia reduces viral load of white spot syndrome virus in Penaeus vannamei

Granja, Clarissa; Vidal, Oscar M.; Parra, Gustavo; Salazar, Marcela

doi:10.3354/dao068175

Cited by 54 publications

(33 citation statements)

References 25 publications

(9 reference statements)

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Section: Resultssupporting

confidence: 80%

“…1B). Non-progression of WSSV infection in crayfish maintained at 32°C is consistent with previous reports of elevated temperatures hindering replication (Du et al 2006, Granja et al 2006, Rahman et al 2006, Gao et al 2011.To examine for DNA fragmentation and morphological changes indicative of cell apoptosis, crayfish tissues were examined using a TUNEL assay and TEM, respectively. In gill and hepatopancreas tissues, no TUNEL-positive cells were detected in uninfected Procambarus clarkii except when tissue was digested with DNase I to purposely fragment DNA as a control ( Fig.…”

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confidence: 53%

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Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature

Wu¹,

Xiong²,

Wang³

et al. 2012

Dis. Aquat. Org.

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In shrimp, higher water temperatures (~32°C) can suppress the ability of white spot syndrome virus (WSSV) to replicate and cause mortality, but the mechanisms remain unclear. To investigate whether cell apoptosis might be involved, a Tdt-mediated dUTP nick-end label (TUNEL) method was used to assess levels of chromosomal DNA fragmentation in hepatopancreas and gill cells of Procambarus clarkii crayfish infected with WSSV and maintained at either 32 ± 1°C or 24 ± 1°C. Based on relative cell numbers with yellow-green colored TUNEL-positive nuclei, the apoptotic index was elevated in WSSV-infected crayfish maintained at 32°C. In gill tissue sections examined by transmission electron microscope, cells with nuclei displaying apoptotic bodies or marginated, condensed and fragmented chromatin without concurrent cell cytoplasm damage were also more prevalent. Flow cytometry sorting of annexin-stained cells showed apoptosis to be most prevalent in granular haemocytes, and assays for caspase-3 activity showed it to be most elevated in hepatopancreas tissue. Despite these indicators of cell apoptosis but consistent with WSSV replication being restricted at elevated temperatures, no increases in transcription of the viral anti-apoptosis genes ORF390 and ORF222 were detected by RT-PCR in shrimp maintained at 32°C, possibly due to the elevated levels of cellular apoptosis. KEY WORDS: WSSV · Crayfish · TUNEL Resale or republication not permitted without written consent of the publisherDis Aquat Org 102: [13][14][15][16][17][18][19][20][21] 2012 Apoptosis is a distinctive mechanism of programmed cell death that can be triggered in response to cell invasion by viruses (O'Brien 1998), thus limiting virus shedding and spread (Barber 2001), and is the main adaptive anti-viral response mounted by terrestrial invertebrates (Koyama et al. 2000). Cell apoptosis has been suggested as a possible mechanism by which Litopenaeus vannamei shrimp survive WSSV infection when maintained at 32°C (Granja et al. 2003). Thus, to investigate this possibility further, approaches to detecting and quantifying cell apoptosis levels including the Tdt-mediated dUTP nick-end label (TUNEL) assay, a caspase-3 activity assay, flow cytometry of annexin-stained haemocytes as well as transmission electron microscopy (TEM) were undertaken on crayfish Procambarus clarkii acclimated to moderate (24°C) or high (32°C) water temperatures before challenge with WSSV. MATERIALS AND METHODS VirusThe WSSV stain used originated from wild Penaeus chinensis collected from the East China Sea in 2001. Using a homogenate of gill tissue from experimentally infected Procambarus clarkii, a semi-purified virus inoculum was prepared using the differential centrifugation method described previously (Du et al. 2007) and stored at −80°C. CrayfishAdult Procambarus clarkii (15−20 g, 8−10 cm long) were purchased from Hangzhou, China, and maintained in 40 l tanks (100 × 50 × 55 cm) containing sand-filtered, ozone-treated, flow-through fresh water. Crayfish were fed with commerc...

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Section: Resultssupporting

confidence: 80%

mentioning

confidence: 53%

Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature

Wu¹,

Xiong²,

Wang³

et al. 2012

Dis. Aquat. Org.

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“…The stimuli used were 1) WSSV infection (early and late times postinfection), 2) injection of heat-killed bacteria and fungal spores, 3) injection of dsRNA, and 4) WSSV infection at 32°C vs. WSSV infection at 27°C. These latter conditions were chosen because hyperthermia (Ͼ31°C) is known to suppress WSSV replication and pathogenicity (20,55).…”

Section: Characterization Of Shrimp Cdna Sequencesmentioning

confidence: 99%

Insights into the immune transcriptome of the shrimpLitopenaeus vannamei: tissue-specific expression profiles and transcriptomic responses to immune challenge

Robalino

Almeida

McKillen

et al. 2007

Physiological Genomics

122

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Infectious disease constitutes a major obstacle to the sustainability of shrimp aquaculture worldwide and a significant threat to natural populations of shrimp and other crustacea. The study of the shrimp immune system, including the response to viral infection, has been hampered by a relative lack of molecular genetic information and of tools suitable for high-throughput assessment of gene expression. In this report, the generation of a cDNA microarray encompassing 2,469 putative unigenes expressed in gills, circulating hemocytes, and hepatopancreas of Litopenaeus vannamei is described. The unigenes printed on the microarray were derived from the analyses of 7,021 expressed sequence tags obtained from standard cDNA libraries as well as from libraries generated by suppression subtractive hybridization, after challenging shrimp with a variety of immune stimuli. The general utility of the cDNA microarray was demonstrated by interrogating the array with labeled RNA from four different shrimp tissues (gills, hemocytes, hepatopancreas, and muscle) and by analyzing the transcriptomic response of shrimp to a lethal challenge with white spot syndrome virus. Our results indicate that white spot syndrome virus infection upregulates (in the hepatopancreas) genes encoding known and potential antimicrobial effectors, while some genes involved in protection from oxidative stress were found to be downregulated by the virus.

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“…25,37). In contrast, work on the relationship between environmental conditions or stressors and susceptibility to viral infections in shrimp is limited, although increasing in recent years, with one report describing the relationship between salinity and infectious hypodermal and hematopoietic necrosis virus (IHHNV) in Litopenaeus vannamei (5), one report describing the effect of acute salinity change on white spot syndrome virus (WSSV) outbreaks in Fenneropenaeus chinensis (40), one report describing the effect of Taura syndrome virus infection on salinity tolerance in L. vannamei (42), a few reports on the effect of water temperature on the pathogenicity (22,24,65) and replication (23) of WSSV in L. vannamei, one report describing the effect of water temperature on IHHNV replication in L. vannamei (45), and one report describing the effect of different types of stress on gill-associated virus (GAV) titer in Penaeus monodon (13).…”

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confidence: 99%

Stress-induced gene expression profiling in the black tiger shrimp Penaeus monodon

Vega

Hall

Wilson

et al. 2007

Physiological Genomics

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Cultured shrimp are continuously exposed to variable environmental conditions that have been associated with stress and subsequent outbreaks of disease. To investigate the effect of environmental stress on Penaeus monodon gene expression, a 3,853 random cDNA microarray chip was generated with clones originating from six stress-enriched hemocyte libraries generated by suppression subtractive hybridization and a normal hemocyte cDNA library. Changes in temporal gene expression were analyzed from shrimp exposed to hypoxic, hyperthermic, and hypoosmotic conditions; 3.1% of the cDNAs were differentially expressed in response to at least one of the environmental stressors, and 72% of the differentially expressed clones had no significant sequence similarity to previously known genes. Among those genes with high identity to known sequences, the most common functional groups were immune-related genes and non-long terminal repeat retrotransposons. Hierarchical clustering revealed a set of cDNAs with temporal and stress-specific gene expression profiles as well as a set of cDNAs indicating a common stress response between stressors. Hypoxic and hyperthermic stressors induced the most severe short-term response in terms of gene regulation, and the osmotic stress had the least variation in expression profiles relative to the control. These expression data agree with observed differences in shrimp physical appearance and behavior following exposure to stress conditions.

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Hyperthermia reduces viral load of white spot syndrome virus in Penaeus vannamei

Cited by 54 publications

References 25 publications

Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature

Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature

Insights into the immune transcriptome of the shrimpLitopenaeus vannamei: tissue-specific expression profiles and transcriptomic responses to immune challenge

Stress-induced gene expression profiling in the black tiger shrimp Penaeus monodon

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