Severe Watery Diarrhea Research Articles

Molecular evolution of porcine epidemic diarrhea virus and porcine deltacoronavirus strains in Central China

Porcine epidemic diarrhea virus (PEDV) and porcine deltacoronavirus (PDCoV) are epizootic swine viruses. To detect and study the evolution of PEDV and PDCoV in central China (Shanxi, Henan, Hubei province), 70 clinical intestinal and fecal samples from piglets with severe watery diarrhea during August 2015 and June 2016 were collected, tested and analyzed. PEDV was more frequently detected by PCR than PDCoV. Phylogenetic analysis of S genes showed that the 10 PEDV strains from this study clustered into G2a (n = 7) and G2b (n = 3) groups. Additionally, the three G2b strains (PEDV S2△) contained the same specific 3 nt deletion in S2 as other reference strains in G2b. Interestingly, complete genome analysis indicated that CH/hubei/2016 was closer to the US INDEL strain and G2a group. CH/hubei/2016 had one recombination event in S2 gene which may have resulted from AH2012-12 (from G2b group) and CH-ZMDZY-11 (from G2a group). Furthermore, 10 purifying selection sites in S gene indicated an adaptive evolution of PEDV in central China swine herds. These results suggested that Pandemic G2a and G2b are predominant PEDV genotype circulating in central China. In addition, the deletion and recombination identified in S gene suggested PEDV strains of central exhibited an evolutionary variety. However, whether these changes affect the pathogenicity and antigenicity of wild PEDV is unknown and is worth for further investigation.

Salmonella Group B and Gastroenteritis Linked to Consumption of Raw Vegetables: A Case Report

We present a case of Salmonella gastroenteritis due to consumption of raw vegetables and water in the farm field.A 62 year-old female from north of Iran was travelling to Karaj. In the way to Karaj, the patient consumed raw unwashed vegetables from the farm field. After a few hours she complained the gastrointestinal disturbances. She admitted to the emergency department for evaluation of frequent severe watery diarrhea. Stool culture was taken at admission time which found positive for Salmonella group B; which was sensitive to Ceftriaxon, Amikacin, Co-trimoxazole, Nalidoxoc acid, Nitrofurantoin, Piperacillin and Ciprofloxacin. Treatment was initiated with Metronidazole and Ciprofloxacin resulting in rapid improvement of the patient’s symptoms. The patient was discharged after 3 days hospital admission. She has remained well without evidence of recurrence of symptoms during a 2-month follow-up period.

Metformin-Associated Lactic Acidosis: A Case Report.

A 54-year-old woman with type 2 diabetes mellitus, hypertension, and peripheral vascular disease developed life-threatening lactic acidosis during treatment with metformin for type 2 diabetes. The woman received metformin at 1000 mg orally twice a day for type 2 diabetes. She presented to our emergency department with a 3-day history of severe watery diarrhea, nausea, and vomiting. Her grandson whom she cared for had gastroenteritis several days prior to the onset of her symptoms. She was confused and hypotensive with a blood pressure of 70/39 mmHg. Her initial laboratory findings were remarkable with an arterial blood gas pH 6.57, HCO _{3}^{ - } 2 mEq/L, anion gap 30 mmol/L, and lactate 16.3 mmol/L. She was diagnosed with severe lactic acidosis. Metformin was discontinued. Upon arrival in the emergency department, she became unresponsive and experienced a pulseless electrical activity cardiac arrest. After resuscitation, her severe acidemia persisted despite aggressive intervention with volume resuscitation and vasopressors, leading to the initiation of renal replacement therapy. After multiple dialysis treatments, her severe acidemia resolved. Serum metformin concentration from presentation ultimately returned to 42 mcg/mL (therapeutic concentration: 1–2 mcg/mL). She was discharged from the hospital on day 15 without any neurologic complications. A Naranjo assessment score of 8 was obtained, indicating a probable relationship between the patient’s lactic acidosis and her use of the suspect drug.

Congenital chloride diarrhea: A rare cause of recurrent polyhydramnios

Congenital chloride diarrhea (CCD) is a rare, inherited disorder. Our case was a preterm neonate who presented with severe watery diarrhoea since Day 2 of life. There was maternal history of polyhydramnios and dilated bowel loops. The diagnosis of CCD was confirmed by mutation analysis. The infant is 9 months corrected age, on salt and potassium supplementation, with age-appropriate milestones. The diagnosis of CCD must be made early to prevent life-threatening fluid and electrolyte imbalance.

Real-time reverse transcription recombinase polymerase amplification assay for rapid detection of porcine epidemic diarrhea virus

Porcine epidemic diarrhea (PED), which is caused by porcine epidemic diarrhea virus (PEDV), is an acute, highly contagious enteric disease characterized by severe watery diarrhea, vomiting, dehydration, and high mortality in suckling piglets. A real-time reverse-transcription recombinase polymerase amplification assay (RT-RPA) was developed based on the nucleocapsid gene of PEDV. RT-RPA assay was performed at 40 °C for 20 min. The assay could detect both the classical and variant PEDV strains, and there was no cross-reaction with other pathogens tested. Using the in vitro transcribed PEDV RNA as template, the analytical sensitivity was 23 copies per reaction. The assay performance was evaluated by testing 76 clinical samples. PEDV RNA positive rate was 55.3% (42/76) by RT-RPA and 59.2% (45/76) by real-time RT-PCR. The diagnostic agreement between the two assays was 96.1% (73/76), and the R2 value of the two assays was 0.903 by linear regression analysis. The developed RT-RPA assay provides a useful alternative tool for simple, rapid and reliable detection of PEDV in resource-limited diagnostic laboratories and on-site facilities.

Vibrio cholerae O1 secretes an extracellular matrix in response to antibody-mediated agglutination.

Vibrio cholerae O1 is one of two serogroups responsible for epidemic cholera, a severe watery diarrhea that occurs after the bacterium colonizes the human small intestine and secretes a potent ADP-ribosylating toxin. Immunity to cholera is associated with intestinal anti-lipopolysaccharide (LPS) antibodies, which are known to inhibit V. cholerae motility and promote bacterial cell-cell crosslinking and aggregation. Here we report that V. cholerae O1 classical and El Tor biotypes produce an extracellular matrix (ECM) when forcibly immobilized and agglutinated by ZAC-3 IgG, an intestinally-derived monoclonal antibody (MAb) against the core/lipid A region of LPS. ECM secretion, as demonstrated by crystal violet staining and scanning electron microscopy, occurred within 30 minutes of antibody exposure and peaked by 3 hours. Non-motile mutants of V. cholerae did not secrete ECM following ZAC-3 IgG exposure, even though they were susceptible to agglutination. The ECM was enriched in O-specific polysaccharide (OSP) but not Vibrio polysaccharide (VPS). Finally, we demonstrate that ECM production by V. cholerae in response to ZAC-3 IgG was associated with bacterial resistant to a secondary complement-mediated attack. In summary, we propose that V. cholerae O1, upon encountering anti-LPS antibodies in the intestinal lumen, secretes an ECM (or O-antigen capsule) possibly as a strategy to shield itself from additional host immune factors and to exit an otherwise inhospitable host environment.

Laboratory Culturing Techniques and Maintenance of Vibrio cholerae.

Cholera is a severe diarrheal disease caused by the consumption of food or water contaminated with the aquatic gram-negative bacterium Vibrio cholerae. Infected hosts will experience vomiting and severe watery diarrhea and if not treated properly will ultimately succumb to death by dehydration. Due to the global prevalence and severity of cholera, V. cholerae has been extensively studied in both environmental and laboratory settings. Herein, we describe proper V. cholerae maintenance, in addition to classical and El Tor biotype culturing in a laboratory setting.

The dynamics of a Chinese porcine G9P[23] rotavirus production in MA-104 cells and intestines of 3-day-old piglets.

Rotavirus A (RVA) G9 genotype is recognized as an emerging genotype which is spreading worldwide, however, our knowledge on pathogenicity of this virus is limited. In this study, porcine RVA strain HN03 was successfully isolated on MA-104 cells, and the isolate was propagated continuously for 7 passages after a virus cloning at passage 3. The virus titers from 4 to 10 passages ranged from 107.1 to 108.1 TCID50/ml. The growth curve of HN03 strain in cell culture was determined, and the virus production dynamics was confirmed by immunoperoxidase monolayer assay (IPMA). Sequence and phylogenetic analyses based on full-length VP7 and partial VP4 genes indicated that HN03 strain belongs to genotype G9P[23]. In addition, the sixth passage of strain HN03 in cell culture was subjected to 3-day-old piglets. All infected piglets developed severe watery diarrhea within 24 hr post-inoculation (hpi), but recovered from disease after 72 hpi. RVA antigen could be detected by IHC in the cytoplasm of villous enterocytes as early as 2 hr after appearance of clinical symptoms and virus antigen load kept increasing in the next 30 hr. The dynamics of RVA HN03 strain proliferation on cells and in pigs extended our understanding of rotavirus pathogenicity.

Nanoparticle-based lateral flow biosensor combined with multiple cross displacement amplification for rapid, visual and sensitive detection of Vibrio cholerae.

Vibrio cholerae is an important human pathogen that is responsible for cholera, a severe acute watery diarrhea. In the current study, a multiple cross displacement amplification (MCDA) coupled with amplicon detection by chromatographic lateral flow biosensor (LFB) method (MCDA-LFB) was successfully established and evaluated for the identification of V. cholerae. A set of 10 primers was designed specifically to recognize 10 different regions of the V. cholerae-specific gene ompW. The optimized time and temperature conditions for the MCDA were 30 min and 63°C, respectively. The MCDA-LFB assay correctly identified 31 strains of V. cholerae but did not detect 13 non-cholerae Vibrio strains and 30 non-Vibrio strains. The sensitivity of MCDA-LFB for target pathogen detection in pure culture was 10 fg per reaction. In the case of spiked shrimp samples without enrichment, the limit of detection was 4.1 CFUs per reaction or equivalent to 4.1×102 CFU g-1. The whole process, including shrimp homogenates processing (30 min), MCDA reaction (30 min) and results reporting (2 min), could be finished within 65 min. These results show that this assay is suitable for the rapid, sensitive and specific detection of V. cholerae in food, environmental and clinical samples.

Development of rapid immunochromatographic strip test for the detection of porcine epidemic diarrhoea virus

Porcine epidemic diarrhoea virus (PEDV) causes acute and severe watery diarrhoea and dehydration, as well as 50–100 per cent mortality in piglets. For the PEDV diagnosis, a rapid test kit...

052 Differential expression of intestinal ion transporters and water channel aquaporins in young piglets challenged with enterotoxigenic Escherichia coli K88

Enterotoxigenic Escherichia coli (ETEC) K88 can induce severe watery diarrhea and subsequent growth retardation for neonatal and postweaning piglets, which causes great economic loss in swine production. Our previous study has shown that the incidence of diarrhea in young piglets was significantly increased following ETEC K88 challenge. The study was to determine whether the expression of genes involved in water and ion transports would be affected by ETEC K88 both in vitro and in vivo. 1) Thirty-six male piglets (4 d old) were randomly allotted to either the Control or the K88 group. Each group had 6 replicates with 3 piglets in each replicate. All piglets were fed the same diets for 18 d. On d 15, piglets in the K88 group were challenged with ETEC K88 (serotype O149:K91:K88ac) of 1 × 108 cfu/pig, whereas those in the Control group received the same volume of sterilized PBS. After challenging with ETEC K88 for 72 h (d 18), 1 piglet from each replicate was selected for slaughter to collect samples from the jejunum, ileum, and colon. The mRNA expression and protein abundance of cystic fibrosis transmembrane conductance regulator (CFTR) in the ileum and colon were significantly increased compared with that in the control group (P < 0.05). Furthermore, the mRNA expression of Na–K–Cl cotransporter 1 (NKCC1) in the ileum and colon was significantly increased, whereas in the jejunum, both its mRNA and protein expression were significantly increased by ETEC K88 treatment (P < 0.05). 2) Additionally, an established porcine intestinal epithelial cell line (IPEC-J2) was used to investigate the effect and possible mechanism of ETEC K88 on expression of water channel aquaporins (AQP) and ion transporters. Cells (1.17 × 106/well) were grown in 6-well plates and treated with ETEC K88 (5.6 × 107 cfu/well) for 3 h. AQP3 and AQP11 mRNA were abundantly expressed in IPEC-J2 cells. The mRNA expression of AQP3, AQP11, and Na+/H+ exchanger 3 (NHE3) in IPEC-J2 cells was significantly reduced after ETEC K88 treatment (P < 0.05). Further analyses by immunofluorescence and western blotting also demonstrated that ETEC K88 significantly decreased the protein expression of AQP3, AQP9, and AQP11 in IPEC-J2 cells (P < 0.05). Moreover, the phosphorylation level of protein kinase A (PKA) of the cyclic adenosine monophosphate (cAMP) pathway was significantly decreased by ETEC K88 challenge (P < 0.05). The results indicate that the cAMP-PKA signaling pathway might regulate the differential expression of intestinal ion transporters and AQP, therefore contributing to fluid imbalance and the development of ETEC diarrhea in young piglets.

Complete Genome Sequences of Porcine Deltacoronavirus Strains DH1/2016 and DH2/2016 Isolated in South Korea.

ABSTRACTTwo porcine deltacoronavirus (PDCoV) strains, named DH1/2016 and DH2/2016, were isolated from feces of piglets which had severe watery diarrhea symptoms. A comparison of the complete genome sequences suggested that the DH1/2016 and DH2/2016 strains are highly homologous to each other and to PDCoVs isolated in early 2014 from the United States.

Transmissible gastroenteritis virus infection induces NF-κB activation through RLR-mediated signaling

Transmissible gastroenteritis virus (TGEV) is a porcine enteric coronavirus which causes lethal severe watery diarrhea in piglets. The pathogenesis of TGEV is strongly associated with inflammation. In this study, we found that TGEV infection activates transcription factors NF-κB, IRF3 and AP-1 in a time- and dose-dependent manner in porcine kidney cells. Treatment with the NF-κB-specific inhibitor BAY11-7082 significantly decreased TGEV-induced proinflammatory cytokine production, but did not affect virus replication. Phosphorylation of NF-κB subunit p65 and proinflammatory cytokine production were greatly decreased after knockdown of retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) or its adaptors MAVS and STING, while only slight reduction was observed in cells following silencing of Toll-like receptor adaptors, MyD88 and TRIF. Furthermore, TGEV infection significantly upregulated mRNA expression of RIG-I and MDA5. Taken together, our results indicate that the RLR signaling pathway is involved in TGEV-induced inflammatory responses.

Racecadotril for the treatment of severe acute watery diarrhoea in children admitted to a tertiary hospital in Kenya

BackgroundDiarrhoea is the second most common cause of death in children under 5 years of age in Kenya. It is usually treated with oral rehydration, zinc and continued feeding. Racecadotril...

First clinical Strongyloides stercoralis case in a dog in Turkey*

A 2.5-year-old small male Cavalier King Charles spaniel was brought to Ondokuz Mayıs University's Faculty of Veterinary Medicine Animal Hospital with symptoms of severe watery diarrhea, pain during defecation, intermittent cough, vomiting, and emaciation. Examination of the fecal smear revealed moderate infection of Strongyloides stercoralis larvae and two adult females. Counting of the larvae in feces was performed by Baermann method and 20 larvae were determined per gram of feces. The dog was treated with ivermectin (Avromec, 0.2 mg/kg subcutaneously) and multivitamin syrup (Vitabiol) for 5 days for supporting the body condition. This paper is the first clinical case report of S. stercoralis in a dog in Turkey.

Successful treatment with fecal microbiota transplantation in patients with multiple organ dysfunction syndrome and diarrhea following severe sepsis.

BackgroundThe dysbiosis of intestinal microbiota plays an important role in the development of gut-derived infections, making it a potential therapeutic target against multiple organ dysfunction syndrome (MODS) after sepsis. However, the effectiveness of fecal microbiota transplantation (FMT) in treating this disease has been rarely investigated.MethodsTwo male patients, a 65-year-old and an 84-year-old, were initially diagnosed with cerebellar hemorrhage and cerebral infarction, respectively, after admission. During the course of hospitalization, both patients developed MODS, septic shock, and severe watery diarrhea. Demographic and clinical data were collected. Intestinal dysbiosis was confirmed by 16S rDNA-based molecular analysis of microbiota composition in fecal samples from the two patients. The two patients each received a single nasogastric infusion of sterile-filtered, pathogen-free feces from a healthy donor. Fecal samples were collected every two days post infusion to monitor changes in microbiota composition in response to treatment.ResultsFollowing FMT, MODS and severe diarrhea were alleviated in both patients. Their stool output and body temperature markedly declined and normalized. Significant modification of microbiota composition, characterized by a profound increase of commensals in the Firmicutes phylum and depletion of opportunistic organisms in the Proteobacteria phylum, was observed in both patients. Furthermore, we identified a reconstituted bacterial community enriched in Firmicutes and depleted of Proteobacteria that was associated with a decrease in the patients’ fecal output and in the levels of plasma inflammation markers.ConclusionsThe outcome of treating two patients with FMT indicates that restoration of the intestinal microbiota barrier can alleviate the infection and modulate the immune response. These findings warrant further investigation of FMT as a putative new therapy for treating microbiota-related diseases such as MODS.

Clostridium Difficile the Landmine of Medicine

Clostridium difficile infection (CDI) is a rising challenge to the US health system. Many intervention measures were designed and implemented in an attempt to control CDI including antibiotic stewardship, improved hand hygiene and contact isolation. We are presenting a case series of 2 patients with a unique infection source. A 69 years old Caucasian female (CF) scrub nurse (patient 1) who participated in an elective ileostomy reversal of a 77 years old CF (patient 2) with past medical history of sigmoidectomy and diverting ileostomy. Patient 1 had complaints of persistent diarrhea for 4 weeks before the operation (initial evaluation at that time was negative for C. diff). She came to the emergency department complaining of severe abdominal pain and watery diarrhea 12 days after the operation. Her stool PCR was positive for C. diff and she was treated with IV metronidazole and oral vancomycin with great response. She was discharged on oral Vancomycin. Patient 2 complained of abdominal pain, loose bowel movement with blood and white count of 19100 two days post-surgery. Her stool PCR was positive for C. diff. Oral vancomycin was started and then switched to IV metronidazole and tigecycline due to her nausea and vomiting. Unfortunately, in the next 3 days her condition deteriorated and as medical intervention failed she had an exploratory laparotomy leading to total abdominal colectomy. She improved and was discharged on oral vancomycin.Figure 1Figure 2Figure 3CDI is one of the major causes of healthcare associated infections. There has been a steady increase in the morbidity and mortality associated with C diff given the advent of more toxigenic/virulent strains. Studies have established asymptomatic carriage in up to two thirds of the patients with C diff colonization. Acquisition of C diff infection can be through feco-oral, surface or skin contamination with spores that are highly resistant to microbicides. Diagnosis is established using the clinical features, EIA of toxin or GDH and PCR in equivocal cases. Patient one complained of diarrhea initially and tested negative for C Diff by EIA (64% sensitive) and the diagnosis was missed. Being a healthcare worker (HCW) and at high risk of acquiring C. Diff/carrier state, is it prudent to treat empirically, do a PCR to screen for C. diff carrier state in HCW's ? With this case series we review the available literature, the accuracy of tests and raise questions on possible interventions and their overall cost effectiveness.

Vibriosis, not cholera: toxigenic Vibrio cholerae non-O1, non-O139 infections in the United States, 1984-2014.

Toxigenic strains of Vibrio cholerae serogroups O1 and O139 have caused cholera epidemics, but other serogroups - such as O75 or O141 - can also produce cholera toxin and cause severe watery diarrhoea similar to cholera. We describe 31 years of surveillance for toxigenic non-O1, non-O139 infections in the United States and map these infections to the state where the exposure probably originated. While serogroups O75 and O141 are closely related pathogens, they differ in how and where they infect people. Oysters were the main vehicle for O75 infection. The vehicles for O141 infection include oysters, clams, and freshwater in lakes and rivers. The patients infected with serogroup O75 who had food traceback information available ate raw oysters from Florida. Patients infected with O141 ate oysters from Florida and clams from New Jersey, and those who only reported being exposed to freshwater were exposed in Arizona, Michigan, Missouri, and Texas. Improving the safety of oysters, specifically, should help prevent future illnesses from these toxigenic strains and similar pathogenic Vibrio species. Post-harvest processing of raw oysters, such as individual quick freezing, heat-cool pasteurization, and high hydrostatic pressurization, should be considered.

Intestinal ameliorative effects of traditional Ogi-tutu, Vernonia amygdalina and Psidium guajava in mice infected with Vibrio cholera.

Cholera, a severe acute watery diarrhea caused by Vibrio cholerae is endemic in Nigeria with most cases occurring in the rural areas. In South West Nigeria, some individuals resort to alternative treatments such as Ogi-tutu, Psidium guajava and Vernonia amygdalina during infections. The effectiveness of these alternatives in the prevention and treatment of V. cholerae infection requires experimental investigation. This study was designed to investigate the ameliorative effects of Ogi-tutu, Vernonia amygdalina and Psidium guajava on intestinal histopathology of experimental mice infected with V. cholerae. Preliminary investigation of in vitro vibriocidal activities of these alternatives were carried out using agar cup diffusion assay. For ameliorative effects, adult mice were inoculated with 100 µl (106 cells) of Vibrio cholerae and dosed at 0 h (immediate prevention) and 4 h (treatment of infection) and their intestines were histopathologically evaluated. The histopathological changes were the same irrespective of the treated groups, but the lesions varied in extent and severity. The ameliorative effects in decreasing order were V. amygdalina > P. guajava > Ogi-tutu. V. amygdalina gave the best ameliorative effects in the prevention and treatment of V. cholerae infection.

Septic Shock Secondary to Salmonella Enterica Ssp. Arizonae in an Immunocompetent Albanian Male

Salmonella-associated diarrhea is a common cause of community-acquired gastroenteritis. Some species of salmonella are associated with invasive diseases like meningitis, endocarditis and septicemia. A 75-year-old Albanian male presented to the emergency department in septic shock. He had history of severe watery diarrhea and high fever of 5 days duration. His stool had a dark green watery consistency imparting a “green pea soup” appearance. Emergency laboratory work-up revealed leukocytosis and acute renal injury. Blood cultures as well as stools yielded S. enterica ssp. arizonae. He was treated with intravenous antibiotics and aggressive fluid resuscitation. Detailed questioning failed to reveal the usual reported risk factors for invasive salmonella infection.