Assessment of fish protein hydrolysate as a substitute for fish meal in white shrimp diets: Impact on growth, immune response, and resistance against Vibrio parahaemolyticus infection.

This study investigated the effects of fish protein hydrolysate derived from barramundi on growth performance, muscle composition, immune response, disease resistance, histology and gene expression in white shrimp (Penaeus vannamei). In vitro studies demonstrated FPH enhanced mRNA expressions of key immune-related genes and stimulated reactive oxygen species (ROS) production and phagocytic activity in shrimp hemocytes. To evaluate the effects of substituting fish meal with FPH in vivo, four isoproteic (43 %), isolipidic (6 %), and isoenergetic diets (489 kcal/100 g) were formulated with fish meal substitution levels of 0 % (control), 30 % (FPH30), 65 % (FPH65), and 100 % (FPH100). After 8-week feeding, the growth performance of FPH65 and FPH100 were significantly lower than that of control and FPH30 (p < 0.05). Similarly, the midgut histological examination revealed the wall thickness and villi height of FPH100 were significantly lower than those of control (p < 0.05). The shrimps were received the challenge of AHPND + Vibrio parahaemolyticus at week 4 and 8. All FPH-fed groups significantly enhanced resistance against Vibrio parahaemolyticus at week 4 (p < 0.05). However, this protective effect diminished after long-period feeding. No significant difference of survival rate was observed among all groups at week 8 (p > 0.05). The expressions of immune-related genes were analyzed at week 4 before and after challenge. In control group, V. parahaemolyticus significantly elevated SOD in hepatopancreas and Muc 19, trypsin, Midline-fas, and GPx in foregut (p < 0.05). Moreover, hepatopancreatic SOD of FPH65 and FPH100 were significantly higher than that of control before challenge (p < 0.05). Immune parameters were measured at week 8. Compared with control, the phagocytic index of FPH 30 was significantly higher (p < 0.05). However, dietary FPH did not alter ROS production, phenoloxidase activity, phagocytic rate, and total hemocyte count (p > 0.05). These findings suggest that FPH30 holds promise as a feed without adverse impacts on growth performance while enhancing the immunological response of white shrimp.

Prophages carrying Zot toxins on different Vibrio genomes: A comprehensive assessment using multilayer networks.

Vibrios, a group of bacteria that are among the most abundant in marine environments, include several species such as Vibrio cholerae and Vibrio parahaemolyticus, which can be pathogenic to humans. Some species of Vibrio contain prophages within their genomes. These prophages can carry genes that code for toxins, such as the zonula occludens toxin (Zot), which contribute to bacterial virulence. Understanding the association between different Vibrio species, prophages and Zot genes can provide insights into their ecological interactions. In this study, we evaluated 4619 Vibrio genomes from 127 species to detect the presence of prophages carrying the Zot toxin. We found 2030 potential prophages with zot-like genes in 43 Vibrio species, showing a non-random association within a primarily modular interaction network. Some prophages, such as CTX or Vf33, were associated with specific species. In contrast, prophages phiVCY and VfO3K6 were found in 28 and 20 Vibrio species, respectively. We also identified six clusters of Zot-like sequences in prophages, with the ZOT2 cluster being the most frequent, present in 34 Vibrio species. This analysis helps to understand the distribution patterns of zot-containing prophages across Vibrio genomes and the potential routes of Zot-like toxin dissemination.

Assessment of Bacillus subtilis fermented Caulerpa microphysa byproduct as feed additive on the growth performance, immune regulation and disease resistance of white shrimp (Litopenaeus vannamei).

In this study, the immunomodulatory and antioxidant activity of fermented Caulerpa microphysa byproduct (FCMB) by Bacillus subtilis was evaluated, and its potential as a feed additive for white shrimp (Litopenaeus vannamei) was explored. In vitro experiments showed that the FCMB supernatant contained polysaccharides, polyphenols and flavonoids, and exhibited antioxidant properties as assessed by various antioxidant assays. Additionally, the FCMB supernatant was found to increase the production rate of reactive oxygen species and the activity of phenoloxidase in hemocytes in vitro. Furthermore, the results of the in vivo feeding trial showed that dietary 5 g kg-1 FCMB significantly improved the weight gain and specific growth rate of white shrimp after 56 days of feeding. Although there were no significant differences in total hemocyte count, phagocytosis, superoxide anion production rate, and phenoloxidase activity among the experimental groups, upregulation of immune-related genes was observed, particularly in the hepatopancreas and hemocytes of shrimps fed with 5 g or 50 g FCMB per kg feed, respectively. In the pathogen challenge assay, white shrimp fed with 5 % FCMB exhibited a higher survival rate compared to the control group following Vibrio parahaemolyticus challenge. Therefore, it is concluded that the fermented byproduct of C. microphysa, FCMB, holds potential as a feed additive for enhancing the growth performance and disease resistance against V. parahaemolyticus in white shrimp.

Quantitative microbial risk assessment of the gastrointestinal risks to swimmers at Southeast Asian urban beaches using site-specific and combined autochthonous and fecal bacteria exposure data.

Recreational exposure to microbial pollution at urban beaches poses a health risk to beachgoers. The accurate quantification of such risks is crucial in managing beaches effectively and establishing warning guidelines. In this study, we employed a quantitative microbial risk assessment (QMRA) framework to assess marine water quality and estimate the risks associated with Vibrio parahaemolyticus, an autochthonous pathogen that causes gastrointestinal illnesses, and enterococci, a traditional fecal bacteria indicator. The microbial contamination levels of V. parahaemolyticus and enterococci were determined from 48 water samples collected at two beaches in Thailand during dry and wet seasons. The accidentally ingested water volumes were obtained through a survey involving 438 respondents. The probability of illness (Pill) was estimated using dose-response models and Monte Carlo simulation. The results revealed that enterococci posed a higher risk of illness than V. parahaemolyticus at all seven study sites. The median combined gastrointestinal (GI) risk from both bacteria at all sites met the US EPA risk benchmark of 0.036 and the 0.05 benchmark set by the WHO, but the 95th percentile risk data at all sites exceeded the benchmarks. This emphasizes the need for the continuous monitoring and management of microbial pollution at these sites. The site-specific exposure data showed higher estimated risks with increased variations compared to the WHO-referenced values, which highlights the significance of locally measured microbial concentrations and survey exposure data to avoid underestimation. Estimating the risks from recreational exposure to waterborne bacteria can inform beach management policies aimed at reducing public health risks to swimmers. The study findings improve the understanding of the risks associated with water recreation activities at Southeast Asian beaches and offer valuable insights for the development of water quality guidelines, which are crucial for the sustainable development of the blue economy.

Inhibitory Effect of Two Closely Related Phages on Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a foodborne pathogenic bacterium commonly found in seafood. The emergence of drug-resistant strains poses a threat to human public health and economic development. Therefore, there are increasing needs to develop new technologies in controlling multidrug-resistant V. parahaemolyticus strains and to evaluate their practical efficiency in seafood or mariculture. In this study, we screened two genetically related V. parahaemolyticus phages, F23s2 and H256D1, which belonged to the siphoviridae family and podoviridae family, respectively. They showed 97.13% and 96.13% identity with Vibrio phage vB_Vpap_MGD1, respectively. Both phages were stable at pH 4-11 and displayed temperature tolerance (<70°C). Meanwhile they showed a broad host spectrum for multidrug-resistant V. parahaemolyticus, and Phage F23s2 lysed 16 of all 23 V. parahaemolyticus strains, while phage H256D1 lysed 10 strains. Phage F23s2 and H256D1 had a good inhibitory effect on V. parahaemolyticus in shrimp meat. Compared with the negative group, the bacterial amount of experimental group with phage F23s2 decreased by 1.60 log colony-forming unit (CFU)/mL at 12 h. For phage H256D1, the bacterial concentration of shrimp meat contaminated with V. parahaemolyticus H256 increased to 5.65 log CFU/mL at 72 h, while the concentration of the experimental group in presence of phage H256D1 was 3.58 log CFU/mL. All live clams infected with V. parahaemolyticus died after 96 h in the absence of phage, whereas clams with phage F23s2 and H256D1 still had a survival rate of 12% and 4%, respectively. Understanding the gene function and biology of phages facilitates its application for control of V. parahaemolyticus contamination worldwide.

Prevalence, Genomic Characterization, and Risk Assessment of Human Pathogenic Vibrio Species in Seafood.

ABSTRACT: Pathogenic Vibrio species are largely responsible for human diseases associated with consumption of contaminated seafood. The aim of this study was to determine the prevalence, population densities, species diversity, and molecular characteristics of pathogenic Vibrio in various seafood commodities and the health risks associated with consumption of these contaminated commodities. Samples of finfish and shellfish (oysters and sea urchins) were collected from various regions and analyzed for Vibrio with the most-probable-number (MPN) technique. Genomic DNA of putative Vibrio isolates was analyzed by whole genome sequencing for taxonomic identification and identification of virulence and antimicrobial resistance genes. The risk of Vibrio-related illnesses due to the consumption of contaminated seafood was assessed with Risk Ranger. Populations of presumptive Vibrio were 2.6 to 4.4 log MPN/g and correlated with season; Vibrio levels were significantly higher (P < 0.05) in the summer. Fifteen Vibrio isolates were identified as Vibrio alginolyticus (five isolates), Vibrio parahaemolyticus (six isolates), Vibrio harveyi (two isolates), and Vibrio diabolicus (two isolates). Two of the six V. parahaemolyticus isolates (ST 2504 and ST 2505) from oysters harbored either the tdh gene for thermostable direct hemolysin or the trh gene for thermostable direct hemolysin-related hemolysin. In addition to virulence genes, the shellfish isolates also harbored genes encoding resistance to multiple antibiotics, including tetracycline, penicillin, quinolone, and ß-lactams, thus arousing concern. The risk assessment predicted that an estimated 21 cases of V. parahaemolyticus-associated gastroenteritis could occur in the general population annually due to consumption of contaminated oysters. This study highlights both the wide prevalence and diversity of Vibrio in seafood and the potential for certain strains to threaten public health.

Enzymatic Specificity of Conserved Rho GTPase Deamidases Promotes Invasion of Vibrio parahaemolyticus at the Expense of Infection.

Vibrio parahaemolyticus is among the leading causes of bacterial seafood-borne acute gastroenteritis. Like many intracellular pathogens, V. parahaemolyticus invades host cells during infection by deamidating host small Rho GTPases. The Rho GTPase deamidating activity of VopC, a type 3 secretion system (T3SS) translocated effector, drives V. parahaemolyticus invasion. The intracellular pathogen uropathogenic Escherichia coli (UPEC) invades host cells by secreting a VopC homolog, the secreted toxin cytotoxic necrotizing factor 1 (CNF1). Because of the homology between VopC and CNF1, we hypothesized that topical application of CNF1 during V. parahaemolyticus infection could supplement VopC activity. Here, we demonstrate that CNF1 improves the efficiency of V. parahaemolyticus invasion, a bottleneck in V. parahaemolyticus infection, across a range of doses. CNF1 increases V. parahaemolyticus invasion independent of both VopC and the T3SS altogether but leaves a disproportionate fraction of intracellular bacteria unable to escape the endosome and complete their infection cycle. This phenomenon holds true in the presence or absence of VopC but is particularly pronounced in the absence of a T3SS. The native VopC, by contrast, promotes a far less efficient invasion but permits the majority of internalized bacteria to escape the endosome and complete their infection cycle. These studies highlight the significance of enzymatic specificity during infection, as virulence factors (VopC and CNF1 in this instance) with similarities in function (bacterial uptake), catalytic activity (deamidation), and substrates (Rho GTPases) are not sufficiently interchangeable for mediating a successful invasion for neighboring bacterial pathogens. IMPORTANCE Many species of intracellular bacterial pathogens target host small Rho GTPases to initiate invasion, including the human pathogens Vibrio parahaemolyticus and uropathogenic Escherichia coli (UPEC). The type three secretion system (T3SS) effector VopC of V. parahaemolyticus promotes invasion through the deamidation of Rac1 and CDC42 in the host, whereas the secreted toxin cytotoxic necrotizing factor 1 (CNF1) drives UPEC's internalization through the deamidation of Rac1, CDC42, and RhoA. Despite these similarities in the catalytic activity of CNF1 and VopC, we observed that the two enzymes were not interchangeable. Although CNF1 increased V. parahaemolyticus endosomal invasion, most intracellular V. parahaemolyticus aborted their infection cycle and remained trapped in endosomes. Our findings illuminate how the precise biochemical fine-tuning of T3SS effectors is essential for efficacious pathogenesis. Moreover, they pave the way for future investigations into the biochemical mechanisms underpinning V. parahaemolyticus endosomal escape and, more broadly, the regulation of successful pathogenesis.

Quantitative Risk Assessment of Vibrio parahaemolyticus in Shellfish from Retail to Consumption in Coastal Cities of Eastern China.

ABSTRACT: Vibrio parahaemolyticus is the main foodborne pathogen worldwide that causes acute gastroenteritis. A quantitative microbiological risk assessment (QMRA) was conducted to evaluate the health risk associated with V. parahaemolyticus in shellfish in the coastal cities in the eastern part of the People's Republic of China. The QMRA framework was established from shellfish at retail to cooking at home to consumption. The prevalence and level of V. parahaemolyticus in shellfish, cooking methods, storage temperature, time after purchase, shellfish consumption frequency, and consumption amount were analyzed in the exposure assessment. The results of the exposure assessment were introduced into the beta-Poisson dose-response model, and Monte Carlo analysis was used to calculate the risk of gastroenteritis from shellfish consumption. The probability of illness caused by V. parahaemolyticus from shellfish consumption per person per year (Pill,yr) was 3.49E-05. Seasonal differences were noted in the Pill/meal; the maximum was 4.81E-06 in summer and the minimum was 2.27E-07 in winter. The sensitivity analysis revealed that the level of V. parahaemolyticus in shellfish and the amount of shellfish consumed per meal were main factors contributing to illness. This QMRA provided valuable information such as the probability of illness associated with the consumption of shellfish and reference points for prevention strategies and control standards of V. parahaemolyticus in shellfish.

Risk Reduction Assessment of Vibrio parahaemolyticus on Shrimp by a Chinese Eating Habit.

In China, a traditional perspective recommended that consuming seafood should be mixed or matched with vinegar, because people thought this traditional Chinese eating habit could reduce the risk of pathogenic microorganism infection, such as Vibrio parahaemolyticus induced diarrhea. However, this empirical viewpoint has not yet been evaluated scientifically. This study conducted a simplified quantitative microbiological risk assessment (QMRA) model, which was employed to estimate the risk reduction of V. parahaemolyticus on ready-to-eat (RTE) shrimp by consuming with vinegars (white vinegar, aromatic vinegar, or mature vinegar). Results showed the reduction of V. parahaemolyticus density on RTE shrimp after consuming with white vinegar, aromatic vinegar and mature vinegar was respectively 0.9953 log CFU/g (90% confidence interval 0.23 to 1.76), 0.7018 log CFU/g (90% confidence interval 0.3430 to 1.060) and 0.6538 log CFU/g (90% confidence interval 0.346 to 0.9620). The infection risk of V. parahaemolyticus per meal in this QMRA model was quantified by a mean of 0.1250 with the standard deviation of 0.2437. After consuming with white vinegar, aromatic vinegar, and mature vinegar, the mean infection risk of V. parahaemolyticus on shrimp decreased to 0.0478, 0.0652, and 0.0686. The QMRA scenarios indicated significant reductions in infection risk when eating RTE shrimp by the Chinese eating habit (consuming with vinegar). This good eating habit should be recommended to promote the spread of around the world.

Assessment of Vibrio parahaemolyticus levels in oysters (Crassostrea virginica) and seawater in Delaware Bay in relation to environmental conditions and the prevalence of molecular markers to identify pathogenic Vibrio parahaemolyticus strains.

This study identified Vibrio parahaemolyticus in oyster and seawater samples collected from Delaware Bay from June through October of 2016. Environmental parameters including water temperature, salinity, dissolved oxygen, pH, and chlorophyll a were measured per sampling event. Oysters homogenate and seawater samples were 10-fold serially diluted and directly plated on CHROMagarᵀᴹ Vibrio medium. Presumptive V. parahaemolyticus colonies were counted and at least 20% of these colonies were selected for molecular chracterization. V. parahaemolyticus isolates (n = 165) were screened for the presence of the species-specific thermolabile hemolysin (tlh) gene, the pathogenic thermostable direct hemolysin (tdh)/ thermostable related hemolysin (trh) genes, the regulatory transmembrane DNA-binding gene (toxR), and V. parahaemolyticus metalloprotease (vpm) gene using a conventional PCR. The highest mean levels of the presumptive V. parahaemolyticus were 9.63×103 CFU/g and 1.85×103 CFU/mL in the oyster and seawater samples, respectively, during the month of July. V. parahaemolyticus levels in oyster and seawater samples were significantly positively correlated with water temperature. Of the 165 isolates, 137 (83%), 110 (66.7%), and 108 (65%) were tlh+, vpm+, and toxR+, respectively. Among the V. parahaemolyticus (tlh+) isolates, 7 (5.1%) and 15 (10.9%) were tdh+ and trh+, respectively, and 24 (17.5%), only oyster isolates, were positive for both genes. Potential pathogenic strains that possessed tdh and/or trh were notably higher in oyster (39%) than seawater (15.6%) isolates. The occurrence of total V. parahaemolyticus (tlh+) was not necessarily proportional to the potential pathogenic V. parahaemolyticus. Co-occurrence of the five genetic markers were observed only among oyster isolates. The co-occurrence of the gene markers showed a relatedness potential of tdh occurrence with vpm. We believe exploring the role of V. parahaemolyticus metalloprotease and whether it is involved in the toxic activity of the thermostable direct hemolysin (TDH) protein can be of significance. The outcomes of this study will provide some foundation for future studies regarding pathogenic Vibrio dynamics in relation to environmental quality.

Affordable automated phenotypic antibiotic susceptibility testing method based on a contactless conductometric sensor.

User-friendly phenotypic antibiotic susceptibility testing (AST) methods are urgently needed in many fields including clinical medicine, epidemiological studies and drug research. Herein, we report a convenient and cost-effective phenotypic AST method based on online monitoring bacterial growth with a developed 8-channel contactless conductometric sensor (CCS). Using E. coli and V. parahaemolyticus as microorganism models, as well as enoxacin, florfenicol, ampicillin, kanamycin and sulfadiazine as antibiotic probes. The minimum inhibitory concentration (MIC) determination was validated in comparison with standard broth microdilution (BMD) assay. The total essential agreements between the CCS AST assays and the reference BMD AST assays are 68.8-92.3%. The CCS has an approximate price of $9,000 (USD). Requiring neither chemical nor biotic auxiliary materials for the assay makes the cost of each sample < $1. The MICs obtained with the automated CCS AST assays are more precise than those obtained with the manual BMD. Moreover, in 72 percent of the counterpart, the MICs obtained with the CCS AST assays are higher than that obtained with the BMD AST assays. The proposed CCS AST method has advantages in affordability, accuracy, sensitivity and user-friendliness.

The human health burden of non-typhoidal Salmonella enterica and Vibrio parahaemolyticus foodborne gastroenteritis in Shanghai, east China.

Information on the burden of disease due to foodborne pathogens in China is quite limited. To understand the incidence of foodborne gastroenteritis due to non-typhoidal Salmonella enterica and Vibrio parahaemolyticus, population survey and sentinel hospital surveillance were conducted during July 2010 to June 2011 in Shanghai, east China, and a model for calculating disease burden was established. The multiplier for gastroenteritis caused by these pathogens was estimated at 59 [95% confidence interval (CI) 30-102]. Annual incidence per 100,000 population in Shanghai was estimated as 48 (95% CI 24-83) and 183 (95% CI 93-317) cases for foodborne non-typhoidal salmonellosis and V. parahaemolyticus gastroenteritis, respectively, illustrating that bacterial gastroenteritis due to these two pathogens poses a substantial health burden. There is a significant difference between our simulated incidence and the data actually reported for foodborne diseases, indicating significant underreporting and underdiagnosis of non-typhoidal S. enterica and V. parahaemolyticus gastroenteritis in the surveillance area. The present research demonstrates basic situation of the health burden caused by major foodborne pathogens in the surveillance area. Enhanced laboratory-based sentinel hospital surveillance is one of the effective ways to monitor food safety in east China.

A database for risk assessment and comparative genomic analysis of foodborne Vibrio parahaemolyticus in China.

Vibrio parahaemolyticus is a major foodborne pathogen worldwide. The increasing number of cases of V. parahaemolyticus infections in China indicates an urgent need to evaluate the prevalence and genetic diversity of this pathogenic bacterium. In this paper, we introduce the Foodborne Vibrio parahaemolyticus genome database (FVPGD), the first scientific database of foodborne V. parahaemolyticus distribution and genomic data in China, based on our previous investigations of V. parahaemolyticus contamination in different kinds of food samples across China from 2011 to 2016. The dataset includes records of 2,499 food samples and 643 V. parahaemolyticus strains from supermarkets and marketplaces distributed over 39 cities in China; 268 whole-genome sequences have been deposited in this database. A spatial view on the risk situations of V. parahaemolyticus contamination in different food types is provided. Additionally, the database provides a functional interface of sequence BLAST, core genome multilocus sequence typing, and phylogenetic analysis. The database will become a powerful tool for risk assessment and outbreak investigations of foodborne pathogens in China.

Assessment of chemical, biological and immunological properties of "Damiana de California" Turnera diffusa Willd extracts in Longfin yellowtail (Seriola rivoliana) leukocytes.

In Mexican herbal medicines or natural remedies, Turnera diffusa (Turneraceae) known as "Damiana de California", has ethnopharmacological relevance, including aphrodisiac, diuretic, and antimicrobial activities. To explore the immunological effect of infusion and methanolic extracts from Damiana de California, this study investigated its chemical, biological, antimicrobial and immunological properties in Longfin yellowtail Seriola rivoliana leukocytes. The analysis of chemical compounds revealed a considerable level of total phenolic and flavonoid contents in the infusion compared with methanolic extract. Furthermore, the antioxidant activity showed high hydroxyl radical scavenging activity in infusion extract compared with BHT positive control. Superoxide radical scavenging activity and ion chelation were higher in methanolic extract followed by infusion treatment. Interestingly, notable antimicrobial activity was observed in both extracts of T. diffusa against Vibrio parahaemolyticus. An in vitro study was performed using leukocytes of S. rivoliana treated with infusion or methanolic extracts at 12.5, 25 and 50 µg/mL for 24 h. Remarkably, infusion extract induced proliferation at any concentration but not the methanolic extract, which was diminished in a dose-dependent fashion. The immunostimulation study demonstrated that the phagocytosis activity increased in those leukocytes stimulated with methanolic extract but diminished the respiratory burst activity, in contrast to the activity observed in those leukocytes stimulated with infusion treatment. Finally, leukocytes incubated with the extracts and confronted with V.parahaemolyticus up-regulated the transcription of proinflammatory cytokine IL-1ß gene in a dose response relationship. These findings suggest that the infusion treatment has potential therapeutic properties, promoting the antioxidant capacity and enhancing immune parameters in Longfin yellowtail S. rivoliana.

Vibrio parahaemolyticus risk assessment in the Pacific Northwest: it's not what's in the water.

The Gram-negative bacterium Vibrio parahaemolyticus (Vp) is a major cause of illness associated with the consumption of raw or undercooked seafood, primarily oysters. This species is a natural member of the bacterial community in brackish waters and is bioaccumulated by oysters through filter feeding. Only a subset of strains is thought to be pathogenic. Currently known virulence markers include the gene for the thermostable direct hemolysin (tdh). In this work we analyzed water and oysters for total Vp and strains encoding tdh from 26 oyster-growing areas of the Puget Sound and Pacific coast of Washington state in 2007 and 2008. In addition, possible plankton-associated Vp were assessed from net tow samples. The density of both total and tdh+ Vp in the water column were considerably higher in 2008 than 2007. However, the concentrations of both total and tdh+ Vp in the oyster tissue was similar for both years. A high proportion of Vp strains in the water column was found to be tdh+ in both 2007 and 2008; however, tdh+ strains were detected at much lower levels in oysters. The data show that analysis of Vp density in the oysters is a better risk assessment tool than density in the overlying water column.

Exposure Assessment and Sensitivity Analysis for Chilled Shrimp During Distribution: A Case Study of Home Delivery Services in Taiwan.

In this study, we evaluated the temperature requirements for home delivery in Taiwan by considering food safety risks of chilled shrimp that may be contaminated with Vibrio parahaemolyticus. Pathogenic V. parahaemolyticus was chosen because it was the main cause of foodborne outbreaks originating from contaminated seafood in Taiwan. The risk of becoming ill due to consumption of raw shrimp was estimated to be 7.3 × 10-3 per serving, estimated based on the real-time temperature profile. Lowering the maximum temperature to 7 °C during transit and cooking shrimps at 100 °C for 5 min could reduce the risk by more than 94%. These interventions, therefore, were suggested to be used as an integral part of temperature management control in the home delivery cold chain. These findings can help food authorities to institute temperature management policies and regulations to prevent broken cold chains and reduce food safety risks. PRACTICAL APPLICATION: This research may be applicable to home delivery services that deliver low-temperature food products, such as seafood products.

Assessment of homeopathic medicines on survival and antioxidant response in white shrimp Litopenaeus vannamei / Evaluación de medicamentos homeopáticos en la supervivencia y respuesta antioxidante del camarón blanco Litopenaeus vannamei infectado con Vibrio parahaemolyticus

ABSTRACT Objective. Evaluating the effect of homeopathic medicines on survival and activity of the superoxide dismutase (SOD) enzyme in white shrimp Litopenaeus vannamei subjected to infection with the pathogenic Vibrio parahaemolyticus. Materials and methods . The average lethal dosage (LD50) was determined for the pathogen strain in L. vannamei juveniles under immersion (Imm), injection (Inj) and incision + immersion (Inc+Imm) methods. Four treatments were applied: (1) Mix CIB®-HOM Heel-Mix (TH1), constituted by equal v/v ratio, of Cyme-Heel®, Gal-Heel®, Hepa-Heel®, Mucs-Heel® and Chol-Heel®; (2) Mix CIB®-HOM Pav-Mix (TH2), constituted by the same v/v ratio of Passiflora incarnata, Valeriana officinalis, Zincum valerianicum and Ignatia amara (Similia®); (3) Heel-Mix/Pav-Mix (TH3) consisting of a 1:1 v/v combination of the TH1 and TH2 treatments, and (4) ViT-Mix (TH4), constituted by Vidatox®, and a control (not treated/infected). Results. While applying the methods Imm, Inj and Inc+Imm, LD50 was 0.9 x 106; 0.6 x 106 and 0.5 x 106 UFC.mL-1, respectively. At the end of the challenge, the groups treated with TH2, TH3 and TH4 had a greater survival rate to that of the control group (p<0.05). Moreover, these two last treatments showed a greater SOD activity with respect to the control group (p<0.05). Conclusions. The homeopathic treatments (TH3 and TH4) increased survival and SOD activity in L. vannamei juveniles challenged with V. parahaemolyticus, which suggests that the homeopathic treatments employed had the potential as an alternative for the control of V. parahaemolyticus and its associated diseases, including the early mortality syndrome in shrimp farming.

RESUMEN Objetivo. Evaluar el efecto de medicamentos homeopáticos sobre la supervivencia y actividad de la enzima superóxido dismutasa (SOD) del camarón Litopenaeus vannamei sometido a infección con Vibrio parahaemolyticus. Materiales y métodos. Se determinó la dosis letal media (DL50) para la cepa patógena en juveniles de L. vannamei, bajo los método de inmersión (Inm), inyección (Iny) e incisión + inmersión (Inc+Inm). Luego el efecto de cuatro medicamentos homeopáticos sobre juveniles de L. vannamei retados con Vibrio parahaemolyticus fue evaluado usando el índice la supervivencia y la actividad SOD. Se aplicaron cuatro tratamientos: (1) Mezcla CIB®-HOM Heel-Mix (TH1), constituido por igual proporción v/v, de Cyme-Heel®, Gal-Heel®, Hepa-Heel®, Mucs-Heel® y Chol-Heel®; (2) Mezcla CIB®-HOM Pav-Mix (TH2), constituido por igual proporción v/v de Passiflora incarnata, Valeriana officinalis, Zincum valerianicum e Ignatia amara (Similia®); (3) Heel-Mix/Pav-Mix (TH3) constituido por una combinación 1:1 v/v de los tratamientos TH1 y TH2, y (4) ViT-Mix (TH4), constituido por Vidatox®, y un control (no tratado/infectado). Resultados. Al aplicar los método Inm, Iny e Inc+Inm la DL50 fue de 0.9 x 106; 0.6 x 106 y 0.5 x 106 UFC.mL-1, respectivamente. Los camarones tratados con TH3 y TH4 presentaron una mayor actividad de SOD con respecto al grupo control (p<0.05). Al final del reto, los grupos TH2, TH3 y TH4 tuvieron una supervivencia mayor a la del grupo control (p<0.05). Conclusiones. Los tratamientos homeopáticos (TH3 y TH4), aumentaron la actividad de la enzima SOD y la supervivencia en juveniles de L. vannamei, retados con V. parahaemolyticus. Esto sugiere que los tratamientos homeopáticos empleados tienen potencial como alternativa para el control de V. parahaemolyticus y sus enfermedades asociadas, incluido el síndrome de mortalidad temprana en el cultivo del camarón.

Simulation of improper food hygiene practices: A quantitative assessment of Vibrio parahaemolyticus distribution.

Kitchen mishandling practices contribute to a large number of foodborne illnesses. In this study, the transfer and cross-contamination potential of Vibrio parahaemolyticus from bloody clams to ready-to-eat food (lettuce) was assessed. Three scenarios were investigated: 1) direct cross-contamination, the transfer of V. parahaemolyticus from bloody clams to non-food contact surfaces (hands and kitchen utensils) to lettuce (via slicing), was evaluated; 2) perfunctory decontamination, the efficacy of two superficial cleaning treatments: a) rinsing in a pail of water, and b) wiping with a kitchen towel, were determined; and 3) secondary cross-contamination, the microbial transfer from cleaning residuals (wash water or stained kitchen towel) to lettuce was assessed. The mean of percent transfer rates through direct contact was 3.6%, and an average of 3.5% of total V. parahaemolyticus was recovered from sliced lettuce. The attempted treatments reduced the transferred population by 99.0% (rinsing) and 94.5% (wiping), and the relative amount of V. parahaemolyticus on sliced lettuce was reduced to 0.008%. V. parahaemolyticus exposure via secondary cross-contamination was marginal. The relative amount of V. parahaemolyticus recovered from washed lettuce was 0.07%, and the transfers from stained kitchen towel to lettuce were insubstantial. Our study highlights that V. parahaemolyticus was readily spread in the kitchen, potentially through sharing of non-food contact surfaces. Results from this study can be used to better understand and potentially raising the awareness of proper handling practices to avert the spread of foodborne pathogens.