The use of quantitative light-induced fluorescence in carious lesions research: A bibliometric review.

OBJECTIVE: This bibliometric analysis evaluated the top 100 most-cited articles on the application of quantitative light-induced fluorescence (QLF) technology in caries research. DATA: The following data were collected: title, authors, country, institution, citations count, title and year of article, study design, topic and keywords. Networks among authors and keywords were constructed by VOSviewer software. SOURCES: Scopus database on April 25, 2024. STUDY SELECTION: A global citation score of 4633 (average 46.33 citations) was calculated with publication years ranged from 1999 to 2020. Caries Research emerged as the top contributing journal. Pretty IA was the most prolific author (18 %). United Kingdom had the highest number of most-cited papers (32 %), followed by Netherlands and USA (20 % each). Laboratory studies constituted the predominant study design (45 %), followed by randomized clinical trials (20 %) and non-systematic reviews (11 %). The keywords "dental caries" and "fluorescence" had 81 and 79 occurrences, respectively. The main topic was QLF use for caries detection (45 %). CONCLUSIONS: This paper provides an update summary of the scientific impact of QLF technology application in caries research. QLF has gained increasing attention worldwide, accompanied by a consistent rise in scientific investigations exploring its application in caries research. CLINICAL SIGNIFICANCE: The findings offer valuable insights into the most influential articles in QLF technology for caries assessment, serving as a critical resource for researchers, clinicians, and students. Understanding the trends in this field can aid in informed decision-making and the advancement of evidence-based practices in caries management and prevention.

Inter-examiner Reliability of Two Methods for Scoring Post-Orthodontic White Spot Lesions from Digital Photographs.

Objective: To compare the reliability of two scoring systems for detecting white spot lesions (WSLs) from clinical photographs captured during debonding of fixed orthodontic appliances. Methods: Digital images of 58 healthy adolescents (34 females and 24 males) were examined, depicting 384 buccal surfaces of maxillary incisors, canines, and first premolars. Three trained examiners (E1, E2 and E3) independently evaluated the fully anonymized photos in a randomized order using the Gorelick index (GI) and the modified International Caries Detection and Assessment System (ICDAS II). A 1-2-week interval separated the scorings. Spearman's rank correlation coefficient, Fisher's z-test, and the interclass correlation coefficient (ICC) were applied to compare the scoring methods and express examiner agreement. Results: The two scoring systems showed a moderate to strong positive relationship, but inter-examiner variations were significant (p<0.05). We found moderate to good reliability (ICC 0.60 to 0.84) with the ICDAS II system and good to excellent values with the GI (ICC 0.72 to 0.94), depending on the examiner. The agreement concerning the sound surfaces and the most severe WSLs was perfect, whereas the scoring of the milder lesion stages appeared more uncertain. Conclusion: A moderate to strong positive relationship was demonstrated between the two methods when scoring the presence and severity of WSLs from digital images. Significant inter-examiner variations affected reliability.

Identification of COP9 signalosome (CSN) subunits and antiviral function analysis of CSN5 in shrimp.

The constitutive photomorphogenesis 9 (COP9) signalosome (CSN) typically composing of eight subunits (CSN1-8) mediates the process of deneddylation and deubiquitination. The fifth subunit of COP9 signalosome, CSN5, has special characteristics compared with the other seven subunits, and plays vital roles in the deneddylation activity and diverse cellular processes. However, the role of CSN5 in antiviral immunity is not clear. In this study, we identified 8 subunits (CSN1-8) of COP9 signalosome in shrimp Marsupenaeus japonicus. CSN1-6 were existed in all tested tissues, but CSN7-CSN8 were not detected in hepatopancreas. After WSSV challenged, the expression level of Csn1 to Csn4, and Csn6 to Csn8 were highly decreased, but the expression level of Csn5 was conspicuously increased in shrimp challenged by white spot syndrome virus (WSSV). The CSN5 was recombinantly expressed in Escherichia coli and its polyclonal antibody was prepared. The expression level of CSN5 was conspicuously increased at RNA and protein levels in the shrimp challenged by WSSV. After knockdown of Csn5 by RNA interference, the WSSV replication was obviously increased in shrimp. When injected the recombinant protein of CSN5 with the membrane penetrating peptide into shrimp, WSSV replication was inhibited and the survival rate of shrimp was significantly improved compared with control. We further analyzed the expression of antimicrobial peptides (AMPs) in Csn5-RNAi shrimp, and the results showed that the expression of several AMPs was declined significantly. These results indicate that CSN5 inhibits replication of WSSV via regulating expression of AMPs in shrimp, and the recombinant CSN5 might be used in shrimp aquaculture for the white spot syndrome disease control.

Litopenaeus vannamei heat shock protein 90 (LvHSP90) interacts with white spot syndrome virus protein, WSSV322, to modulate hemocyte apoptosis during viral infection.

As cellular chaperones, heat shock protein can facilitate viral infection in different steps of infection process. Previously, we have shown that the suppression of Litopenaeus vannamei (Lv)HSP90 not only results in a decline of white spot syndrome virus (WSSV) infection but also induces apoptosis in shrimp hemocyte cells. However, the mechanism underlying how LvHSP90 involved in WSSV infection remains largely unknown. In this study, a yeast two-hybrid assay and co-immunoprecipitation revealed that LvHSP90 interacts with the viral protein WSSV322 which function as an anti-apoptosis protein. Recombinant protein (r) LvHSP90 and rWSSV322 inhibited cycloheximide-induced hemocyte cell apoptosis in vitro. Co-silencing of LvHSP90 and WSSV322 in WSSV-infected shrimp led to a decrease in expression level of viral replication marker genes (VP28, ie-1) and WSSV copy number, while caspase 3/7 activity was noticeably induced. The number of apoptotic cells, confirmed by Hoechst 33342 staining assay and annexin V/PI staining, was significantly higher in LvHSP90 and WSSV322 co-silenced-shrimp than the control groups. Moreover, the co-silencing of LvHSP90 and WSSV322 triggered apoptosis by the mitochondrial pathway, resulting in the upregulation of pro-apoptotic protein expression (bax) and the downregulation of anti-apoptotic protein expression (bcl, Akt). This process also involved the release of cytochrome c (CytC) from the mitochondria and a decrease in mitochondrial membrane potential (MMP). These findings suggest that LvHSP90 interacts with WSSV322 to facilitate viral replication by inhibiting host apoptosis during WSSV infection.

Remineralization potential of varying concentrations of two plant-based extracts of Cocos nucifera on white spot lesions using SEM and EDAX analysis: An in vitro study.

Aim: To evaluate the remineralization potential of varying concentrations of two plant-based extracts of Cocos nucifera on white spot lesions using SEM and EDAX in vitro testing methods. Materials and Methods: The pulp was freshly obtained from coconut and divided into two. Then, coconut milk was obtained by blending, while the next portion was freeze-dried and lyophilized. Third molar teeth were processed into tooth slabs (N = 40) and split equally into five groups by block randomization. After demineralization, one tooth slab was taken from each, and SEM analysis was done. Remineralization was then performed among the various groups that included Group 1, which acted as a control and consisted of the remineralization solution. Groups 2 and 3 comprised 1:1 and 2:1 concentrations of the coconut milk, whereas Groups 4 and 5 consisted of 1:1 and 2:1 concentrations of the lyophilized extract. SEM and EDAX testing were done post-remineralization. Ca and phosphate values were tabulated, and statistical significance was determined for the obtained values using ANOVA. Results: Among the control and treatment groups, surface remineralization was better observed in 1:1 coconut milk and 2:1 coconut milk than in the 2:1 lyophilized coconut, control, and 1:1 lyophilized coconut. Between the control and treatment groups, Ca and phosphate percentages (P < 0.001) showed statistical differences. The lowest value of 2.3% was noted in the 2:1 lyophilized coconut group. Conclusion: Coconut extracts exhibit remineralization potential on the artificial carious lesion. Coconut milk exhibited significant improvement in the surface properties than lyophilized coconut.

The genome sequence of the Marbled White Spot, Protodeltote pygarga (Hufnagel, 1766).

We present a genome assembly from an individual male Protodeltote pygarga (the Marbled White Spot; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence is 421.1 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.48 kilobases in length. Gene annotation of this assembly on Ensembl identified 17,784 protein coding genes.

Development of an Artificial White Spot Lesion Creation Protocol: A Preliminary Study.

BACKGROUND AND OBJECTIVES: Protocols that determine the lesion depth of specific demineralization solutions are lacking. This in vitro study aimed to evaluate various lesion depths of artificial white spot lesions (WSLs) at different exposure times.  Materials and methods: Artificial WSLs were created by placing 30 extracted human premolar teeth into 0.05M acetate buffer solution with a controlled environment of pH 4.4 at 37ºC, which were then exposed in the solution for various durations of 4, 5, 6, 8, 10, and 12 days. The specimens were visually examined using the Ekstrand-Ricketts-Kidd (ERK) system to confirm the WSL, followed by buccolingual sectioning, and evaluated under a scanning electron microscope (SEM) to measure the lesion depth. RESULTS: The SEM showed that the mean lesion depths of representative specimens were 101.33 µm (day 4), 124 µm (day 5), 159 µm (day 6), 187 µm (day 8), 202 µm (day 10), and 212 µm (day 12). The artificial WSL was visually demonstrated in grades 1 and 2 of the ERK system. CONCLUSIONS: The depths of the lesions increased as the duration increased from day 4 to day 12, indicating that the lesion depths increased with the more prolonged exposure to the acetate buffer solution.

Efficacy of F-ACP-Containing Dental Mousse in the Remineralization of White Spot Lesions after Fixed Orthodontic Therapy: A Randomized Clinical Trial.

Fixed appliance (FA) therapy predisposes patients to white spot lesions (WSLs). The F-ACP complex (amorphous calcium phosphate nanoparticles enriched with carbonate and fluorine and coated with citrate) has been effective for in vitro enamel remineralization. The aim of this study was to evaluate the efficacy of the F-ACP complex in remineralizing WSLs after FA therapy. One hundred and six adolescents (aged 12-20 years) were randomized into study and control groups after FA therapy. Patients in the study group were advised to use dental mousse containing F-ACP applied within Essix retainers for six months. The presence of WSLs was recorded at baseline (T0), 3 months (T1), and 6 months (T2) according to the International Caries Detection and Assessment System (ICDAS). Visual Plaque Index (VPI) and Gingival Bleeding Index (GBI) were recorded. Among 106 study participants, 91 (52 and 39 in study and control groups, respectively) completed the study. The results showed that the ICDAS score was significantly lower (p < 0.001) in the study group than in the control group between T0 and T2. The application of mousse containing the F-ACP complex inside Essix retainers for six months is effective in remineralizing white spot lesions in patients after FA therapy without side effects.

Caries and white spot lesion trajectories of orthodontic patients across an observation period of 20 years.

OBJECTIVES: Fixed orthodontic appliances may increase the risk for caries and white spot lesions. The aim of this retrospective study was to determine the long-term associations between both in orthodontic patients. MATERIALS AND METHODS: 103 patients aged 36.6 ± 6.5 years whose fixed appliance orthodontic treatment had finished at least 15 years ago were included. Current clinical data and photographs (T3), panoramic x-ray and photographs from before treatment (T0), after debonding (T1) and at 2-year follow-up (T2) were available. Parameters of interest were dentine caries, "Missing/Filled Teeth" (MFT), "White Spot Lesion" (WSL) index and "Periodontal Screening and Recording" index (PSR; T3 only). RESULTS: At T0, 30.4% had no caries experience decreasing to 25.6%, 22.4% and 6.8% at T1, T2 and T3 resp. The median MFT (95% CI) at T0, T1, T2 and T3 was 2 (1;3), 3 (2;4), 3 (2;4) and 7 (6;9) resp. increasing significantly at each time point (p < 0.001 each); 30.1% had WSL at debonding. Patients with caries experience at T0 had a 2.4-fold increased risk of WSL at debonding. Dentine caries, caries experience and WSL at T1 were significantly associated with incident caries at T2, but not at T3. PSR at T3 had a significant association with previous caries incidence and WSL. CONCLUSIONS: Caries experience prior to orthodontic treatment may constitute a risk indicator for WSL, and caries experience and WSL at its end for caries incidence in the near term. CLINICAL RELEVANCE: Present caries and WSL may help identifying orthodontic patients with special need for prevention and counselling.

The Effect of Non-Invasive Treatment Techniques on the Color Masking Ability and Surface Roughness of Induced Enamel Lesions (An in vitro Study).

Aim of the Study: The aim of this in vitro study was to evaluate the effect of the resin infiltration technique and remineralization of induced enamel caries with fluoride solution on the color masking of white spot lesions and surface roughness. Materials and Methods: A total of 45 sound teeth were used in this study. All the teeth were sectioned along the long axes into two halves, lingual and buccal to get 90 specimens. All specimens were immersed in a demineralized solution for 7 days. The specimens were divided randomly into three equal groups (n = 30) according to the type of treatment; Group 1 (n = 30) was treated with fluoride varnish (Clinpro); Group 2 (n = 30) was treated by resin infiltration (Icon); and Group 3 (n = 30) was used as a control group with no treatment. The color and surface roughness were measured three times at baseline (T1), directly after induction of artificial white spot lesions (WSLs) (T2), and directly after application of the treatment options (T3). The colors were measured using a portable reflective spectrophotometer and the 3D surface roughness was measured using a Light Sectioning Vision System. Data were collected and statistically analyzed using T test and Mann-Whitney U test. The Results: Surface roughness was almost equal in the study groups with no statistically significant differences reported. Icon showed slightly higher color scores than that of ClinPro. Conclusions: The Icon produced favorable esthetic results compared to the fluoride therapy while no significant differences were reported regarding the surface roughness.

A Peritrophin-44 gene in Litopenaeus vannamei is involved in disease resistance.

Peritrophic membrane (PM) is a pellicle structure present in the midgut of some invertebrates, such as insects and crustaceans. It could isolate harmful components and pathogens in food from intestinal epithelial cells; and it also plays a role in improving digestion and absorption efficiency. So PM is important for survival of its owner. In current study, 44 PM proteins were identified in Litopenaeus vannamei by PM proteome analysis. Among these PM proteins, the Peritrophin-44 homologous protein (LvPT44) was further studied. Chitin-binding assay indicated that LvPT44 could bind to colloidal chitin, and immunoeletron microscopy analysis shown that it was located to PM of L. vannamei. Furthermore, LvPT44 promoter was found to be activated by L. vannamei STAT and c-Jun. Besides, LvPT44 was induced by ER-stress as well as white spot syndrome virus infection. Knocked-down expression of LvPT44 by RNA inference increased the cumulative mortality of shrimp that caused by ER-stress or white spot syndrome virus. These results suggested that LvPT44 has an important role in disease resistance.

Evaluation of resin infiltration for inhibiting initial caries progression: An in vitro study using Micro-Computed Tomographic analysis.

Objective: To evaluate the changes in lesion depth and mineral density of resin infiltration-treated white spot lesions against a simulated oral environment using thermal and acidic challenges in vitro. Materials and methods: Two enamel slabs were prepared from each buccal surface of permanent human premolars, for a total of 56 slabs. Artificial white spot lesions were induced. One specimen was treated with resin infiltration, while the other was used as an untreated control. A micro-CT was used to assess the lesion depth and mineral density of each specimen. Subsequently, all specimens were subjected to 10,000 cycles of thermocycling and pH cycling for 10 days before being re-evaluated using the micro-CT. Lesion depth and mineral density were examined and compared between before and after aging procedures within each group by the paired sample t-test. The independent samples t-test was utilized to compare lesion depth progression and percentage change of mineral density between groups. Results: After aging, there was both a significant lesion depth progression and a mineral loss in the control and resin infiltration groups. Mean lesion depth progression was 132.88 ± 4.18 µm for the control group and 52.31 ± 4.16 µm for resin infiltration group. Percentage mineral density loss as a percentage for the control and resin infiltration groups were 16.1 ± 0.64 % and 8.83 ± 0.30 %, respectively. The resin infiltration group demonstrated a significantly lower mean lesion depth progression and percentage changes in mineral loss compared to the control group. Conclusions: The lesion depth and mineral density changes in the resin infiltrated-treated group were lower than untreated white spot lesions after aging procedures using thermal and acidic challenges. Clinical significance: Resin infiltration is a promising approach to inhibit the progression of white spot lesions related to the initial stage of dental caries.

White spot lesions in fixed orthodontic treatment: Etiology, pathophysiology, diagnosis, treatment, and future research perspectives.

White spot lesions (WSLs) refer to localized areas of hypo-mineralization limited to the enamel of the teeth surface, which is noticeable clinically to the naked eye on drying of the teeth. During fixed orthodontic treatment, it is very hard for the patient to maintain excellent oral hygiene as the brackets, bands, wires, elastics, and other appliances and attachments worn intra-orally provide a platform for food retention, plaque formation, and then colonization by acidogenic bacteria like Streptococcus mutans and Lactobacillus. This review aims to elaborate and focus on etiology, pathophysiology, diagnosis, treatment aspect, and future scope for research about the WSLs occurring due to fixed orthodontic treatment.

Effectiveness of fluoride mouthrinse in prevention of demineralization during fixed orthodontic treatment: A systematic review and meta-analysis.

Enamel demineralization is a very common occurrence around bonded brackets in an orthodontic practice. Fluoride (FLR) applications have been used to prevent decalcification and further progression of white spot lesions. The purpose of this systematic review and meta-analysis was to systematically appraise available literature on the effectiveness of fluoride mouthrinse in the prevention of demineralization around fixed orthodontic appliances. A search was conducted for randomized controlled clinical trials among four electronic databases (MEDLINE, Google Scholar, PubMed, and Cochrane Review) through MeSH terms and keywords. Studies were excluded if random allocation was not conducted, or if they were animal or in vitro studies. About 146 articles were screened and 5 studies were selected for the present review. Only two studies were selected for MA due to variations in the measurement of outcomes among studies. This review concluded that rinsing with FLR in the course of the fixed orthodontic treatment lessens demineralization around the bracket. Using FLR mouthrinse to inhibit the formation of white spot lesions or dental caries in patients with multiple cavities or restoration can be considered in clinical practice.

A Modern Multidisciplinary Method to Characterize Natural White Spot Lesions with 2D and 3D Assessments: A Preliminary Study.

In this preliminary study, a multidisciplinary method based on high-resolution analytical techniques (such as microcomputed tomography, Raman Microspectroscopy, scanning electron microscopy, and Vickers microhardness test) was exploited to evaluate the alterations that occur in human teeth at the initial stage of the carious lesion. To this purpose, six extracted molars displaying a natural white spot lesion (WSL) were investigated. Specific morphological, structural, and chemical parameters, such as the mineral density, indentation hardness, molecular and elemental composition, and surface micromorphology were obtained on the WSL, and the results were statistically compared (t-test, p < 0.05) to those of the sound enamel on the same tooth. In the WSL, with respect to the sound area, a decrease in the mineral density and crystallinity was detected together with differences in the molecular composition and surface microstructure, such as the occurrence of micropores and irregularities. Moreover, the elemental analysis highlighted in WSL showed a statistically significant decrease in Ca and P percentages. In conclusion, this multidisciplinary approach allows us to fully characterize the area of interest, providing a deeper knowledge of these enamel lesions, which could have important clinical implications.

Evaluation of incipient enamel-carious-like lesion treated with hydroxyapatite-chitosan nanocomposite hydrogel.

BACKGROUND: Non-invasive restoration of tooth enamel is a topic of high relevance in dental material science. Multiple approaches have been proposed to reach optimum reconstruction results. The current study was performed to evaluate the cross-sectional microhardness besides mineral quality and content in artificially induced carious enamel after treatment with hydroxyapatite-chitosan (HAp-CS) nanocomposite gel. METHODS: Artificially carious lesions were induced by immersion of teeth in acidic carboxymethyl cellulose gel (pH 4.95-5) for 24- and 72-hours periods. Two different compositions of HAp-CS nanocomposite hydrogel were prepared with two different ratios 50/50 (%) and 70/30 (%), respectively. Additionally, sodium fluoride gel (1000 ppm concentration) was prepared and used as reference. Gels were applied to carious lesions twice/day for 3 min/each. After 45 days of application, surface morphology, energy dispersive x-ray spectroscopy, micro-Raman analysis in addition to cross-sectional microhardness were evaluated. Statistical analysis was performed using two-way ANOVA and Tukey's post hoc statistical tests. RESULTS: Surface morphological evaluation of treated surfaces showed obliteration of surface irregularities. Groups demineralized for 24 hours and treated with 70/30 (HAp-CS) showed highest significant cross-sectional-microhardness (P ≤ 0.05). Evaluated subsurface cross-sectional microhardness showed better mineral quality for groups demineralized for 24 hours and treated with HAp-CS nanocomposite gels. CONCLUSIONS: Nanocomposite gel with 70/30 (HAp-CS) could efficiently improve cross-sectional microhardness and both minerals composition and quality for lesions demineralized for 24 hours. More severely induced lesions, as demineralized for 72 hours, need more powerful agent compositions and/or prolonged application protocols for improvement.

Resin infiltration versus fluoride varnish for visual improvement of white spot lesions during multibracket treatment. A randomized-controlled clinical trial.

AIMS: This study aimed to evaluate the visual improvement of resin infiltration of white spot lesions (WSL) during orthodontic treatment with the multibracket appliance (MBA) compared to fluoride varnish. METHODS: Patients aged 12-17 years with at least one WSL with an International Caries Detection and Assessment System (ICDAS) score of 1-2 during an active MBA treatment were included and randomized to receive either resin infiltration (Icon) or fluoride application (Flairesse). Standardized digital images were obtained before, one-day, one-week, one-month, three-months and six-months after treatment using a DSLR camera and a matching polarization filter. A grey reference card was used for color standardization. A Matlab routine was used to measure the color difference between adjacent healthy enamel and treated WSL. The independent-samples t-test was used for intergroup and paired-samples t-test for intragroup comparison. RESULTS: Images of 116 teeth from 36 patients were analyzed. The ΔE for the "Icon" treated WSL was smaller (T1ICON = 5.0 ± 1.4) than in the fluoride group (T1Fluoride = 8.4 ± 3.2). Caries infiltration significantly improved the aesthetic appearance of WSL (p < 0.001), which remained satisfactory at six months (T5ICON = 5.2 ± 1.6). CONCLUSION: WSL infiltration management during orthodontic treatment was superior to topical fluoridation in not only arresting the enamel lesions but also significantly improving the aesthetic appearance of demineralized regions around the brackets. CLINICAL RELEVANCE: WSL treatment in orthodontic patients is usually initiated after debonding. Research has shown that the earlier WSL is treated, the better the aesthetic outcome. There is limited data on the efficacy of resin infiltration of WSL during orthodontic treatment.

Evaluation of the surface characteristics and antibacterial properties of Titanium dioxide nanotube and methacryloyloxyethylphosphorylcholine (MPC) coated orthodontic brackets-a comparative invitro study.

OBJECTIVES: White spot lesions are the most common iatrogenic effect observed during orthodontic treatment. This study aimed to compare the surface characteristics and antibacterial action of uncoated and coated orthodontic brackets. MATERIALS AND METHODS: Sixty commercially available stainless steel brackets were coated with TiO2 nanotubes and methacryloyloxyethylphosphorylcholine. The sample was divided into Group 1: uncoated orthodontic brackets, Group 2: Stainless steel brackets with TiO2 nanotubes coating, Group 3: Stainless steel brackets with methacryloyloxyethylphosphorylcholine coating, and Group 4: Stainless steel brackets with TiO2 nanotubes combined with methacryloyloxyethylphosphorylcholine coating. Surface characterization was assessed using atomic force microscopy and scanning electron microscopy. Streptococcus mutans was selected to test the antibacterial ability of the orthodontic brackets, total bacterial adhesion and bacterial viability were assessed. The brackets were subjected to scanning electron microscopy to detect the presence of biofilm. RESULTS: The surface roughness was the greatest in Group 1 and least in Group 2 followed by Group 4 and Group 3 coated brackets. The optical density values were highest in Group 1 and lowest in Group 4. Comparison of colony counts revealed high counts in Group 1 and low counts in Group 4. A positive correlation between surface roughness and colony counts was obtained, however, was not statistically significant. CONCLUSIONS: The coated orthodontic brackets exhibited less surface roughness than the uncoated orthodontic brackets. Group 4 coated orthodontic brackets showed the best antibacterial properties. CLINICAL RELEVANCE: Coated orthodontic brackets prevent adhesion of streptococcus mutans and reduces plaque accumulation around the brackets thereby preventing formation of white spot lesions during orthodontic treatment.

WSSV early protein WSSV004 enhances viral replication by suppressing LDH activity.

White spot syndrome virus (WSSV) is known to upregulate glycolysis to supply biomolecules and energy for the virus's replication. At the viral genome replication stage, lactate dehydrogenase (LDH), a glycolytic enzyme, shows increased activity without any increase in expression. In the present study, yeast 2-hybrid screening was used to identify WSSV proteins that interacted with LvLDH isoform 1 and 2, and these included the WSSV early protein WSSV004. The interaction between WSSV004 and LvLDH1/2 was confirmed by co-immunoprecipitation. Immunofluorescence showed that WSSV004 co-localized with LvLDH1/2 in the cytoplasm. dsRNA silencing experiments showed that WSSV004 was crucial for WSSV replication. However, although WSSV004 silencing led to the suppression of total LvLDH gene expression during the viral late stage, there was nevertheless a significant increase in LvLDH activity at this time. We also used affinity purification-mass spectrometry to identify cellular proteins that interact with WSSV004, and found a total of 108 host proteins and 3 WSSV proteins with which it potentially interacts. Bioinformatics analysis revealed that WSSV004 and its interacting proteins might be responsible for various biological pathways during infection, including vesicular transport machinery and RNA-related functions. Collectively, our study suggests that WSSV004 serves as a multifunctional modulator to facilitate WSSV replication.

Experimental Infection Models and Their Usefulness for White Spot Syndrome Virus (WSSV) Research in Shrimp.

White spot syndrome virus (WSSV) is marked as one of the most economically devastating pathogens in shrimp aquaculture worldwide. Infection of cultured shrimp can lead to mass mortality (up to 100%). Although progress has been made, our understanding of WSSV's infection process and the virus-host-environment interaction is far from complete. This in turn hinders the development of effective mitigation strategies against WSSV. Infection models occupy a crucial first step in the research flow that tries to elucidate the infectious disease process to develop new antiviral treatments. Moreover, since the establishment of continuous shrimp cell lines is a work in progress, the development and use of standardized in vivo infection models that reflect the host-pathogen interaction in shrimp is a necessity. This review critically examines key aspects of in vivo WSSV infection model development that are often overlooked, such as standardization, (post)larval quality, inoculum type and choice of inoculation procedure, housing conditions, and shrimp welfare considerations. Furthermore, the usefulness of experimental infection models for different lines of WSSV research will be discussed with the aim to aid researchers when choosing a suitable model for their research needs.