Diarrhea In Humans Research Articles

Pet Ownership Protects Against Recurrence of Clostridioides difficile Infection.

Background Clostridioides difficile infection (CDI) is the leading cause of antibiotic-associated and health care–associated diarrhea in humans. Recurrent CDI (R-CDI) occurs in ~20%–30% of patients with CDI and results in increased morbidity, mortality, and hospital costs. Genomic analyses have shown overlap of C. difficile isolates from animals and people, suggesting that a zoonotic reservoir may contribute to recurrence. The objective of this study was to determine whether pet ownership is a risk factor for recurrence of CDI.MethodsWe conducted a case–control study among patients with recurrent CDI (cases; n = 86) and patients with nonrecurrent CDI (controls; n = 146). Multivariable logistic regression modeling was used to determine the association between recurrence of CDI and pet ownership while accounting for patient-level risk factors.ResultsPet ownership was not significantly associated with recurrence of CDI (odds ratio [OR], 1.02; 95% confidence interval [CI], 0.38–2.72; P = 0.965) among all patients (n = 232). However, among the subset of patients with community-associated or community-onset health care facility–acquired CDI (n = 127), increasing contact with pets was increasingly protective against recurrence: for every point increase in a pet contact score (out of 7 possible points), the odds of recurrence decreased by 14% (OR, 0.86; 95% CI, 0.74–1.00; P = 0.051).ConclusionsClose interactions with pets appear protective against the recurrence of community-acquired CDI. A potential mechanism may involve beneficial contributions to the microbiota of pet owners afflicted with CDI, as has been observed for other conditions such as atopy, obesity, and food allergies. However, more research is needed to understand the interactions between pets, owners, and their microbiota.

Group A Rotavirus Genotypes among Calf Herds in Southwest Nigeria and Implications for Human Rotavirus Vaccines’ Efficiency

Rotaviruses have been widely reported to be associated with diarrhea in humans but fewer studies abound on other mammalian species. This prospective study was conducted to detect and characterize Rotaviruses from freely ranged migratory herds of cattle in Ekiti and Ondo states, Nigeria with a view to further expanding knowledge on rotaviruses, possible animal-human interspecies transmission and impacts on vaccine efficiency. By convenience sampling, between September 2014 and February 2015, stool samples from 120 calves, comprising settled and migratory herds of cattle were obtained and examined for group A rotaviruses using Certest® Quadruple Enzyme Immuno Assay. Rotavirus genomes were isolated by extraction from the positive samples, reverse transcribed and amplified by One-Step reverse transcription polymerase chain reaction (RT-PCR), and genotyped by semi-nested multiplex PCR. Representative PCR products of the genotyped samples were purified and sequenced using Sanger method. The generated query sequences were queried to the GenBank to retrieve similar sequences for pairwise alignment by ClustalW. Phylogenetic analyses by Neighbour-joining method were conducted at 1000 bootstrap replicates to obtain phylograms using MEGA 6 software. Fifteen samples (15/120: 12.5%) were positive for rotavirus. No statistically significant association existed between rotavirus infection and gender of the calves (χ2 = 0, df = 1, p = 1). Eight samples (8/15: 53.3%) were successfully genotyped where the G1, G5, G9, G10, G12, and P[6] were detected. Nucleotide Sequencing of the VP7 and VP4 genes of the genotyped samples confirmed strains G1P[6], G10P[6], and G12P[6] with 58% - 100% nucleotide identity within these viruses. Phylogenetic analysis revealed 2 possible transmissions from India and Honduras. Bovine rotaviruses were detected in freely ranged and settled calf herds in southwestern Nigeria at a rate of 12.5%. The strains identified were analysed to be group A rotavirus strains with potential interspecies transmission from human to calves and from bovine to humans. The detected mixed strains could eventually impact negatively on the effectiveness of available rotavirus vaccines over the prevailing serotypes in human infections.

Differential antimicrobial susceptibility profiles between symptomatic and asymptomatic non-typhoidal Salmonella infections in Vietnamese children.

Non-typhoidal Salmonella (NTS) serovars, sequences types and antimicrobial susceptibility profiles have specific associations with animal and human infections in Vietnam. Antimicrobial resistance may have an effect on the manifestation of human NTS infections, with isolates from asymptomatic individuals being more susceptible to antimicrobials than those associated with animals and human diarrhoea.

Escherichia coli in Chicken Carcasses in Southern Brazil: Absence of Shigatoxigenic (STEC) and Isolation of Atypical Enteropathogenic (aEPEC)

ABSTRACT The aim of this study was to investigate the presence of shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) in frozen chicken carcasses sold at stores in southern Brazil. Typical E. coli colonies were enumerated in 246 chicken carcasses, and the presence of stx1, stx2, eae genes was investigated in their rinse liquid and in E. coli strains isolated from those carcasses. Strains of E. coli were also investigated for the presence of bfp gene. A median of 0.6 cfu.g-1(ranging from <0.1 to 242.7 cfu.g-1) of typical E. coli colonies was found in the carcasses. Shiga toxin-encoding genes (stx1 and stx2) were not detected, indicating that the chicken carcasses were negative for STEC. The intimin protein gene (eae) was detected in E.coli isolated from 4.88% of the carcasses; all tested strains were negative for the bfp gene and were classified as aEPEC. Twenty-two aEPEC strains were tested for resistance to ten antimicrobials and subjected to macrorestriction (PFGE). All the tested aEPEC strains were fully susceptible to cephalosporins, ciprofloxacin and colistin. Resistance to sulfonamide (65%), ampicillin (55%), tetracycline (50%) and gentamicin (45%) were the most frequent. The PFGE profile demonstrated a low level of similarity among the resistant strains, indicating that they were epidemiologically unrelated. The results indicate that aEPEC strains can contaminate chicken meat, and their association with strains implicated in human diarrhea needs to be further investigated.

Update on Cryptosporidium spp.: highlights from the Seventh International Giardia and Cryptosporidium Conference.

While cryptosporidiosis is recognized as being among the most common causes of human parasitic diarrhea in the world, there is currently limited knowledge on Cryptosporidium infection mechanisms, incomplete codification of diagnostic methods, and a need for additional therapeutic options. In response, the Seventh International Giardia and Cryptosporidium Conference (IGCC 2019) was hosted from 23 to 26 June 2019, at the Rouen Normandy University, France. This trusted event brought together an international delegation of researchers to synthesize recent advances and identify key research questions and knowledge gaps. The program of the interdisciplinary conference included all aspects of host-parasite relationships from basic research to applications to human and veterinary medicine, and environmental issues associated with waterborne parasites and their epidemiological consequences. In relation to Cryptosporidium and cryptosporidiosis, the primary research areas for which novel findings and the most impressive communications were presented and discussed included: Cryptosporidium in environmental waters, seafood, and fresh produce; Animal epidemiology; Human cryptosporidiosis and epidemiology; Genomes and genomic evolution encompassing: Comparative genomics of Cryptosporidium spp., Genomic insights into biology, Acquiring and utilizing genome sequences, Genetic manipulation; Host-parasite interaction (immunology, microbiome); and Diagnosis and treatment. High quality presentations discussed at the conference reflected decisive progress and identified new opportunities that will engage investigators and funding agencies to spur future research in a “one health” approach to improve basic knowledge and the clinical and public health management of zoonotic cryptosporidiosis.

Update on Giardia: Highlights from the seventh International Giardia and Cryptosporidium Conference.

Although Giardia duodenalis is recognized as one of the leading causes of parasitic human diarrhea in the world, knowledge of the mechanisms of infection is limited, as the pathophysiological consequences of infection remain incompletely elucidated. Similarly, the reason for and consequences of the very specific genome-organization in this parasite with 2 active nuclei is only partially known. Consistent with its tradition, the 7th International Giardia and Cryptosporidium Conference (IGCC 2019) was held from June 23 to 26, 2019, at the Faculty of Medicine and Pharmacy of the University of Rouen-Normandie, France, to discuss current research perspectives in the field. This renowned event brought together an international delegation of researchers to present and debate recent advances and identify the main research themes and knowledge gaps. The program for this interdisciplinary conference included all aspects of host-parasite relationships, from basic research to applications in human and veterinary medicine, as well as the environmental issues raised by water-borne parasites and their epidemiological consequences. With regard to Giardia and giardiasis, the main areas of research for which new findings and the most impressive communications were presented and discussed included: parasite ecology and epidemiology of giardiasis, Giardia-host interactions, and cell biology of Giardia, genomes and genomic evolution. The high-quality presentations discussed at the Conference noted breakthroughs and identified new opportunities that will inspire researchers and funding agencies to stimulate future research in a “one health” approach to improve basic knowledge and clinical and public health management of zoonotic giardiasis.

Origin and Evolution of Hybrid Shiga Toxin-Producing and Uropathogenic Escherichia coli Strains of Sequence Type 141.

Hybrid Shiga toxin-producing Escherichia coli (STEC) and uropathogenic E. coli (UPEC) strains of multilocus sequence type 141 (ST141) cause both urinary tract infections and diarrhea in humans and are phylogenetically positioned between STEC and UPEC strains. We used comparative genomic analysis of 85 temporally and spatially diverse ST141 E. coli strains, including 14 STEC/UPEC hybrids, collected in Germany (n = 13) and the United States (n = 1) to reconstruct their molecular evolution. Whole-genome sequencing data showed that 89% of the ST141 E. coli strains either were STEC/UPEC hybrids or contained a mixture of virulence genes from other pathotypes. Core genome analysis and ancestral reconstruction revealed that the ST141 E. coli strains clustered into two lineages that evolved from a common ancestor in the mid-19th century. The STEC/UPEC hybrid emerged ∼100 years ago by acquiring an stx prophage, which integrated into previously unknown insertion site between rcsB and rcsD, followed by the insertion of a pathogenicity island (PAI) similar to PAI II of UPEC strain 536 (PAI II536-like). The two variants of PAI II536-like were associated with tRNA genes leuX and pheU, respectively. Finally, microevolution within PAI II536-like and acquisition of the enterohemorrhagic E. coli plasmid were observed. Our data suggest that intestinal pathogenic E. coli (IPEC)/extraintestinal pathogenic E. coli (ExPEC) hybrids are widespread and that selection pressure within the ST141 E. coli population led to the emergence of the STEC/UPEC hybrid as a clinically important subgroup. We hypothesize that ST141 E. coli strains serve as a melting pot for pathogroup conversion between IPEC and ExPEC, contrasting the classical theory of pathogen emergence from nonpathogens and corroborating our recent phenomenon of heteropathogenicity among pathogenic E. coli strains.

Comparable stx2a expression and phage production levels between Shiga toxin-producing Escherichia coli strains from human and bovine origin.

Shiga toxin-producing Escherichia coli (STEC) can cause diarrhoea and severe diseases in humans, such as haemolytic uraemic syndrome. STEC virulence is considered to correlate with the amount of Shiga toxins (Stx) produced, especially Stx2, whose subtype Stx2a is most frequently associated with high virulence. Stx are encoded in prophages, which play an important role in STEC pathogenesis. The aim of this study was to evaluate stx2a expression levels and Stx2a phage production using qPCR and the double-agar-layer method in 29 STEC strains, corresponding to serotypes O26:H11 (6), O91:H21 (1), O145:H- (11) and O157:H7 (11), isolated from cattle and humans. Results were then tested for possible associations with serotype, origin or some genetic features. We observed heterogeneous levels of stx2a expression and Stx2a phage production. However, statistical comparisons identified a higher stx2a expression in response to mitomycin C in strains isolated from cattle than in those from humans. At the same time, compared to stx2a /stx2c strains, stx2a strains showed a higher increase in phage production under induced conditions. Notably, most of the strains studied, regardless of serotype and origin, carried inducible Stx2a phages and evidenced expression of stx2a that increased along with phage production levels under induced conditions.

DAV131A Protects Hamsters from Lethal Clostridioides difficile Infection Induced by Fluoroquinolones.

Fluoroquinolone treatments induce dysbiosis of the intestinal microbiota, resulting in loss of resistance to colonization by exogenous bacteria such as Clostridioides difficile that may cause severe diarrhea in humans and lethal infection in hamsters. We show here that DAV131A, a charcoal-based adsorbent, decreases the intestinal levels of the fluoroquinolone antibiotics levofloxacin and ciprofloxacin in hamsters, protects their intestinal microbiota, and prevents lethal infection by C. difficile.

BIOCIDAL POTENTIALS OF STEM BARK EXTRACTS OF PSIDIUM GUAJAVA (LINN.) ON PANEL OF BACTERIAL STRAINS ASSOCIATED WITH DIARRHEA AND DYSENTERY

Objective: Investigations were carried out on cidal effects of crude extract and fractions obtained from stem bark of Psidium guajava against a panel of 74 strains of Escherichia coli and Shigella dysenteriae implicated in diarrhea and dysentery infections.&#x0D; Methods: Powdered sample of the stem bark was extracted in methanol/distilled water (3:2) and then partitioned into different organic solvents. The fractions obtained were subjected to antibacterial tests against a panel of bacterial strains. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined using agar dilution method while the mode of action of the active fractions was investigated through time-kill dynamics and leakages of proteins and potassium ions from the cells.&#x0D; Results: All partitioned fractions except the aqueous fraction exhibited antibacterial activities against the panel of bacterial strains at a final concentration of 10 mg/mL. The MIC exhibited by the crude extract against susceptible bacterial strains ranged between 1.56 mg/mL and 12.5 mg/mL, while the MIC for the four fractions ranged between 0.31 mg/mL and 5.0 mg/mL. The MBC ranged between 3.13–12.5 mg/mL and 0.63–5.0 mg/mL for the crude extract and fractions, respectively. The time-kill assay revealed that the percentage of the cells killed increase with an increase in the concentrations of the fractions as well as contact time intervals. Proteins and potassium ions leakages from the bacterial cells followed the same trend with that of time-kill assay.&#x0D; Conclusion: The stem bark extracts of P. guajava exhibited appreciable bactericidal effects on bacterial strains associated with diarrhea and dysentery in humans.

Ethnopharmacological Potentials of Warburgia ugandensis on Antimicrobial Activities.

Warburgia ugandensis (W. ugandensis) is known by various names, including the East African greenheart, pepper bark tree, and Ugandan greenheart, and has a rich history of extensive use in the treatment of a host of human diseases in many African countries. This review is based on the botany and ethnopharmacological potentials of W. ugandensis for the treatment of pneumonia, asthma, malaria, candidiasis, skin infections, human immunodeficiency virus opportunistic infections, diarrhea, and measles given the common use in the management of these diseases. Extracts from W. ugandensis have strong antimicrobial activities against a broad spectrum of pathogens mainly because of the presence of abundant terpenoids, drimane, and coloratane type sesquditerpenoids amongst which are ugandensial, warburganal, mukaadial, and other secondary metabolites, such as tannins, flavonoids, saponins, steroids, and mannitol. This group of compounds gives the plant a high therapeutic value. Based on the review, there is a need for identification and isolation of the highly therapeutic phytochemical constituents and a drive for more preclinical and clinical trials to validate the safety and efficacy of the extracts. This gives basis for the potential development of new therapeutic drugs from the plant.

Isolation and toxin gene detection of Clostridium (Clostridioides) difficile from traditional and commercial quail farms and packed quail meat for market supply - Short communication.

Clostridium (Clostridioides) difficile is a Gram-positive anaerobic rod-shaped bacterium and the main cause of nosocomial diarrhoea in humans. In recent years, the transmission of C. difficile from environmental reservoirs (e.g. food) to humans has become a major focus of research. The aim of this study was to investigate the prevalence and corresponding toxin genes of C. difficile in faecal samples and meat of quails. Thirty samples of packed quail meat in Mashhad, Iran and 500 faecal samples (pooled to n = 5) were collected on quail farms in the Northeastern Khorasan region for further investigation. Of 100 pooled quail faecal samples 10% showed cultural growth of C. difficile. In meat samples two out of 30 specimens (7%) showed cultural growth. In six of ten isolates from faecal samples toxin genes (tcdB and tcdA) were present, while four isolates harboured no toxin genes. However, in meat isolates no toxin genes were present. Mutations in the tcdC gene were not detected, indicating that 'hypervirulent' strains such as RT027 and RT078 were not present. The data suggest that quail and quail products might hold a potential for the spread of C. difficile.

Porcine rotavirus mainly infects primary porcine enterocytes at the basolateral surface

Intestinal epithelium functions as a barrier to protect multicellular organisms from the outside world. It consists of epithelial cells closely connected by intercellular junctions, selective gates which control paracellular diffusion of solutes, ions and macromolecules across the epithelium and keep out pathogens. Rotavirus is one of the major enteric viruses causing severe diarrhea in humans and animals. It specifically infects the enterocytes on villi of small intestines. The polarity of rotavirus replication in their target enterocytes and the role of intestinal epithelial integrity were examined in the present study. Treatment with EGTA, a drug that chelates calcium and disrupts the intercellular junctions, (i) significantly enhanced the infection of rotavirus in primary enterocytes, (ii) increased the binding of rotavirus to enterocytes, but (iii) considerably blocked internalization of rotavirus. After internalization, rotavirus was resistant to EGTA treatment. To investigate the polarity of rotavirus infection, the primary enterocytes were cultured in a transwell system and infected with rotavirus at either the apical or the basolateral surface. Rotavirus preferentially infected enterocytes at the basolateral surface. Restriction of infection through apical inoculation was overcome by EGTA treatment. Overall, our findings demonstrate that integrity of the intestinal epithelium is crucial in the host’s innate defense against rotavirus infection. In addition, the intercellular receptor is located basolaterally and disruption of intercellular junctions facilitates the binding of rotavirus to their receptor at the basolateral surface.

Effects of stachyose on intestinal microbiota and immunity in mice infected with enterotoxigenic Escherichia coli

Abstract Enterotoxigenic Escherichia coli (ETEC) is an important pathogen causing diarrhea in humans. Finding natural, safe and effective functional ingredients to replace antibiotics is an important direction for preventing and treating E. coli infection and protecting intestinal health. However, it is unknown whether stachyose promotes the proliferation of probiotics and thus inhibits ETEC infection. Here, we investigated whether stachyose improves the gut microbiota of ETEC-infected mice and induces effects in early postnatal life. Stachyose promoted the proliferation of A. muciniphila and Bifidobacterium, increased short chain fatty acids (SCFAs) levels, increased the expression of tight junction proteins (occludin and ZO-1), increased SIgA and albumin levels, and reduced the level of lipopolysaccharide (LPS), IL-1, 6, 17 and TNF-α. After antibiotic-induced ablation, the resistance of the intestinal microbiota of the ETEC-infected mice to bacteria was significantly reduced, and the level of inflammation was increased, the SIgA and albumin levels decreased significantly, showed the intestinal microbiota plays an important role in maintaining immune resistance. Stachyose enabled partial microbiota recovery and protection after ETEC infection.

Conocimientos, actitudes y prácticas acerca de parasitosis intestinal en adultos que asistieron a consulta de atención primaria en la comunidad de Paya municipio Baní, provincia Peravia, República Dominicana durante el periodo mayo-junio 2018

Introducción: el parasitismo intestinal es un problema de gran prevalencia a nivel de salud pública mundial, responsable de aproximadamente el 10 % de las diarreas en humanos. Las parasitosis intestinales son infestaciones producidas por parásitos cuyo hábitat natural es el aparato digestivo de las personas y animales. Es una de las principales causas de consulta en las clínicas rurales. También, es una de las más importantes causas de malnutriciones en poblaciones pediátricas.&#x0D; Material y métodos: este estudio fue de carácter prospectivo, descriptivo y de corte transversal. Para adquirir la información discutida se empleó un protocolo (ver sección de anexos) basado en una investigación previa y adaptada por los autores. Se tomaron en cuenta las fichas familiares de los pacientes.&#x0D; La población estudiada corresponde a todos los pacientes de Paya Arriba que asistieron a la consulta en la Unidad de Atención Primaria de Paya. Como criterio de inclusión se requirió que la persona fuese mayor de edad, y que se identificase a sí mismo como cabecera del hogar, residente permanente de la comunidad de Paya Arriba y que voluntariamente participara en el estudio.&#x0D; Conclusión: el 60 % de los entrevistados está consciente de que tiene un nivel intermedio de conocimientos, ya que se autoevaluaron a sí mismos con un tres de cinco, siendo el cinco el mayor puntaje. Mientras que un 28 % se autoevaluó con dos de cinco, lo que pone de manifiesto que está consciente de la necesidad de educación en el tema.&#x0D; Se exhorta a los habitantes a recalcar lo imprescindible de acciones tan sencillas como el lavado de manos para promover salud, para promover vida.

Evaluation of thermal destruction kinetics of Clostridium difficile spores (ATCC 17857) in lean ground beef with first‐order/Weibull modeling considerations

AbstractThermal destruction kinetics of spores of Clostridium difficile ATCC 17857 was evaluated between 74 and 82°C and characterized using the first‐order log‐linear and Weibull models. Computed decimal reduction times using the first‐order model ranged from 4.39 min at 82°C to 146 min at 74°C, with a z value of 5.17°C. Thermal destruction data were also analyzed using the Weibull model. Based on regression, the predicted one‐D value (first‐order model) and the reliable life (tR) (Weibull model) were 3.86 and 4.05 min at 82°C and 136 and 165 min at 74°C, respectively, indicating the Weibull model to be more conservative yielding higher decimal reduction time values. However, when extended to achieve 2.5, 4, and 6 decimal reductions in C. difficile spores, the calculated process times were more conservative with the first‐order model than with the Weibull model. Moreover, within the experimental range, when data for both models could be compared, predictions from the first‐order model were much closer to the experimental values. Therefore, when used for process calculation for 2.5 or higher log reductions, the first‐order model would give more conservative and safer process times. The study provides destruction kinetics data for C. difficile under a range of temperature conditions.Practical ApplicationsClostridium difficile is a major cause of antibiotic‐associated diarrhea and pseudomembranous colitis in humans. C. difficile infection is the leading cause of gastroenteritis‐associated death. C. difficile infections have been increasing in recent years, and therefore warrant appropriate remedial measures. Thermal inactivation is the most common method for pathogen control, and often the cooking practices are adjusted to a level pre‐established to make the foods pathogen free. Available information on thermal destruction kinetics is scarce, and therefore the data generated here should be of significant importance for safety considerations.

Cryptosporidium and giardia species in newly and previously habituated gorillas and nearby water sources in Bwindi Impenetrable National Park, Uganda.

Introductioncryptosporidium and giardia are of great one health significance and major cause of protozoan diarrhea in humans and primates; they are found in the faecal matter of animals and humans and also in contaminated water and soil as well. Therefore, we aimed at establishing the prevalence and shedding intensity of faecal Cryptosporidium and giardia in the Newly Habituated Mountain Gorillas (NHMG) and Previously Habituated Mountain Gorillas (PHMG) and in selected water sources within the gorilla home ranges in the month of January 2018.Methodswe conducted a cross sectional study in the southern sector of Bwindi Impenetrable National Park where a total of 56 faecal samples from both NHMG (34) and PHMG (22) and 30 water samples were purposively collected. Faecal and water samples were transported in a cooler box at 4ºC to Makerere University Parasitology Laboratory for analysis. The samples were analyzed using modified Ziehl-Neelsen technique and Ether concentration method for Cryptosporidium and giardia respectively.Resultsthe prevalence of cryptosporidium was established as 13 (59.1%), 15 (44.1%) and 7 (23.3%) in PHMG, NHMG and water respectively. The mean concentration of the oocysts per gram was 222±52.9 in PHMG, 174±41.5 in NHMG and 31±13.2 in water. The prevalence of giardia was 3 (13.6%), 4 (11.8%) and 3 (10%) in PHMG, NHMG and water respectively. The mean concentration of the oocysts per gram was 34±19.9 in PHMG, 25±12.4 in NHMG and 5±2.9 in water. There was no significant difference in both the prevalence of cryptosporidium (p>0.05) and giardia (p>0.05) in the PHMG and NHMG. This indicates that there is high risk of cross infection among the gorillas within the forest sharing similar home ranges.Conclusionthe park authorities should ensure that procedures for proper waste disposal while in the forest are properly followed, water drawn for drinking from the forest should be avoided. Further research should be carried out to identify whether the strains of the parasites found in water or other animals in the forest are the same with strains in gorilla dung in order to confirm cross infection.

Prevalence and molecular characterization of Cryptosporidium and Giardia in pre-weaned native calves in the Republic of Korea.

Cryptosporidium spp. and Giardia duodenalis are protozoan parasites that cause diarrhea in humans and animals. Molecular data on Cryptosporidium spp. and G. duodenalis in calves in the Republic of Korea (ROK) is limited; thus, we investigated the prevalence of Cryptosporidium and Giardia in pre-weaned calves, analyzed the association between these parasites and diarrhea, and identified their zoonotic potential for human infection. Fecal samples were collected from 315 pre-weaned calves aged 1-60 days from 10 different regions in the ROK and screened for Cryptosporidium spp. and G. duodenalis using PCR. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 4.4% (n = 14) and 12.7% (n = 40), respectively. Co-infection was not detected. All Cryptosporidium-positive samples were identified as C. parvum after sequence analysis of a small subunit rRNA fragment and further subtyped into zoonotic IIaA15G2R1 (n = 13) and IIaA18G3R1 (n = 1) by DNA sequencing of the 60-kDa glycoprotein gene. To our knowledge, this is the first report of C. parvum IIaA15G2R1 subtype in calves in the ROK. Based on β-giardin (bg) gene, G. duodenalis-positive samples belonged to assemblages E (n = 36) and A (n = 4), with the latter belonging to subtype A1, the zoonotic genotype. Six subtypes of assemblage E were identified at the bg locus: E1 (n = 6), E2 (n = 3), E3 (n = 13), E5 (n = 1), E8 (n = 1), and E11 (n = 1). The occurrence of C. parvum and G. duodenalis was not associated with diarrhea in pre-weaned Korean native calves. The present results suggest that the prevalence of C. parvum is not related to calf age; in contrast, the prevalence of G. duodenalis was significantly higher in 41-50-day-old calves (odds ratio = 9.90, 95% confidence interval 2.37-41.34; P = 0.001) than in 1-10-day-old calves. Therefore, calves are a potential source of zoonotic transmission, which may have significant public health implications.

A new emerging serotype of Vibrio parahaemolyticus in China is rapidly becoming the main epidemic strain

ObjectivesDuring surveillance, we found a new type of Vibrio parahaemolyticus named ‘O4:KUT-recAin’ and studied the phenotypic, pathogenic and epidemiological characteristics of O4:KUT-recAin. MethodsV. parahaemolyticus were isolated from acute diarrhoeal patients in coastal hospitals of China. Serum agglutination test, specific PCR assay, growth curves under different conditions and rabbit diarrhoeal models were using to characterize O4:KUT-recAin. ResultsThe O4:KUT-recAin strain has a new type of K antigen and a 25 043-bp-large fragment encoding 20 proteins inserted in the housekeeping gene recA. Retrospective analysis found that only one O4:KUT-recAin strain was detected in 563 V. parahaemolyticus strains in 2014; then the proportion increased rapidly and reached 17.8% (105/590) in 2016 and 31.1% (224/721) in 2017, making O4:KUT-recAin the second dominant serotype following O3:K6. O4:KUT-recAin strains (100%, 14/14) exhibited increased acid resistance and could reproduce in medium at pH 4.9, while 92.9% (13/14) of the O3:K6 strains could not grow at this pH value. O4:KUT-recAin could cause diarrhoea and small intestinal tissue lesions in infant rabbits, but its diarrhoeal (93.1%, 27/29) and mortality (78.6%, 22/28) rates were slightly lower than those of O3:K6 (100% 16/16, 100% 16/16). Based on diarrhoea patients, there were no significant differences in the two groups for most clinical symptoms and laboratory results, except media age, haemoglobin and the number of red blood cells in stool samples. ConclusionsO4:KUT-recAin had enhanced acid resistance, was capable of causing infectious diarrhoea in both rabbits and humans, and has become widespread during a short period of time in China.

Comparison of oral, intravenous, and subcutaneous fluid therapy for resuscitation of calves with diarrhea

Neonatal diarrhea remains the primary cause of mortality in dairy calves around the world, and optimal treatment protocols are needed. The main goals of therapy are to restore hydration and electrolyte concentrations, correct strong ion (metabolic) acidemia, and provide nutritional support. Administration of oral electrolyte solutions (OES) has long been the primary method used to treat neonatal diarrhea in humans and calves because OES are capable of addressing each of the primary goals of therapy. In calves with moderate dehydration, we hypothesized that oral electrolytes would be as good as or better than small volumes of intravenous (IV) or subcutaneous (SC) fluids. Therefore, the main goal of this study was to compare the ability of a commercially available oral electrolyte solution (OES) administered alone or in combination with hypertonic saline with small volumes of IV or SC fluid therapy to resuscitate calves with diarrhea. Thirty-three Holstein calves from 5 to 14 d of age were utilized in this clinical trial. Diarrhea and dehydration were induced by adding sucrose to the milk replacer. In addition, hydrochlorothiazide and spironolactone were given orally and furosemide intramuscularly. Depression status, clinical hydration scores, fecal consistency, and body weight were recorded at regular intervals. Treatment began when calves had severe diarrhea and had a decrease in plasma volume of at least 10%. Calves were randomly assigned to 1 of 4 treatment groups of 8 to 9 calves per group: (1) OES; (2) OES with hypertonic saline (4 mL/kg, IV); (3) IV fluids (lactated Ringer's, 2 L); or (4) SC fluids (lactated Ringer's, 2 L). Treatments were given at 0 and 12 h. Changes in plasma volume, blood pH, electrolyte levels, and physical examination scores were determined before therapy and again at 1, 2, 4, 8, and 12 h after each treatment. All 4 treatments were ultimately successful in improving hydration as well as increasing blood pH; however, animals in both groups that received OES had much faster resuscitation than those in either the IV or SC fluid group. In conclusion, oral electrolyte products remain the gold standard for resuscitating diarrheic calves with moderate dehydration and acidemia and will likely perform better than small volumes of IV lactated Ringer's solution. Subcutaneous fluids by themselves are a poor treatment option and should be only be used as supportive therapy following the initial correction of hypovolemia and metabolic acidosis.