Diarrhea Mortality Research Articles

The Relationship of Diarrhea in COVID-19 Patients With Transaminitis, Severity, and Mortality

Background: Coronavirus disease 2019 (COVID-19) is a pandemic that causes various types of symptoms. Diarrhea is a gastrointestinal symptom that is hypothesized to be associated with patient morbidity and mortality. Research on the relationship between diarrhea and the incidence of transaminitis, severity and mortality has never been carried out, so further research is neededMethod: Descriptive-analytic research design with a case-control study approach. Sampling using consecutive sampling on patients in the inpatient installation of dr. Saiful Anwar, Malang. Research data on diarrhea incidence, transaminitis, severity, and mortality were obtained from secondary data on COVID-19 patients. Data were analyzed using chi-square with a significance level p 0.05Results: The incidence of diarrhea in COVID-19 patients with transaminitis was 45.8% and significantly associated (p = 0.025). Diarrhea in COVID-19 patients was associated with severity (p = 0.046) and patient mortality (p = 0.028).Conclusion: There is a relationship between the incidence of diarrhea in COVID-19 patients with trasnsaminitis, severity and mortality

In vitro antibacterial activity of extracts of Dalbergia sissoo and Aegle marmelos against Enterotoxigenic Escherichia coli from calves

Neonatal calf diarrhoea (NCD) is the most common cause of morbidity and mortality in pre-weaned dairy calves, causing huge economic and productivity losses to dairy industry worldwide. Enterotoxigenic E. coli (ETEC) is one of the important causes of neonatal diarrhoea and high mortality in very young calves. Shisham (Dalbergia sissoo) leaves and Bael (Aegle marmelous) unripe fruit have been used in diarrhoea and dysentery with very good results. Hence the present study was conducted to evaluate the in vitro antibacterial activity of the aqueous and methanolic extract of Dalbergia sissoo and Aegle marmelos against 16 E. coli isolated from colibacillosis affected diarrhoeic calves in Bikaner district of Rajasthan. The average zones of inhibitions of aqueous extract of Aegle marmelos and methanolic extract of Dalbergia sissoo against E. coli were 23.75±0.34 mm, 18.75±0.32 mm, 16.5±0.24 mm, 11.58 ±0.14 mm and 7.88±0.24 mm at concentrations of 1000 mg/ml, 500 mg/ml, 250 mg/ml 125 mg/ml and 62.5 mg/ml, respectively. The average zones of inhibition of aqueous extract of Dalbergia sissoo and methanolic extract of Aegle marmelos against E. coli were 24.75±0.20 mm, 19.87±0.34 mm, 17.25±0.24mm, 12.86±0.13 mm and 8.50±0.14 mm at concentrations of 1000 mg/ml, 500 mg/ml, 250 mg/ml 125 mg/ml and 62.5 mg/ml, respectively. The maximum zone of inhibition was reported by aqueous extract of Dalbergia sissoo and methanolic extract of Aegle marmelos (>24 mm of diameter) at concentration of 1000 mg/ml. Based on average zone of inhibition, the in vitro antibacterial activity of aqueous extract of Dalbergia sissoo and methanolic extract of Aegle marmelos was found to be more against E. coli as compared to aqueous extract of Aegle marmelos and methanolic extract of Dalbergia sissoo at varying concentrations. In conclusion, both combination of aqueous and methanolic extract of Dalbergia sissoo and Aegle marmelos showed in vitro antibacterial properties against E. coli.

Pre-weaned dairy calf management practices, morbidity and mortality of bovine respiratory disease and diarrhea in China

Controlling disease, maintaining animal well-being, and sustaining production efficiency in Chinese dairy industry will depend on whether enough of China's dairy farmers can adopt proper calf management practices. Consequently, to provide insights into a number of beneficial management practices used by many Chinese dairy farmers, ninety-eight dairy farms, located in the country's three typical climate zones (severe cold zone, SCZ; cold zone, CZ; and hot-summer/cold-winter zone, HSCWZ) were surveyed. The various management practices employed and the morbidity and mortality rates caused by bovine respiratory disease and diarrhea were recorded at each investigated farm. About three quarters of the calves on these farms were housed indoors (either individually or in group pens). The proportion of the calves reared in separate pens (individual indoor or outdoor hutches) was highest (56.20%) at large-scale farms (> 500 cows), followed by 33.3% at medium-scale farms (50–500 cows). A majority of the investigated farms (≥ 68.04%) did not have a system for monitoring either temperature, relative humidity, or air quality. A medium depth (score 2) of bedding was most common at 42.11% of the farms. 87.76% of the farms relied on natural ventilation. 36.10% of the farms did not provide supplemental heating in the calf barn, and the heating methods most often used differed among the three climate zones. Manual milk feeders involving bottles or buckets were more common (83.67%) than automated milk feeders. The mean morbidity and mortality rates of pre-weaned calves infected with bovine respiratory disease were 11.73% (95% CI:10.2–13.23%) and 3.65% (95% CI: 2.85–4.45%), while those infected with diarrhea were 14.17% (95% CI: 12.46–15.88%) and 3.61% (95% CI: 2.97–4.25%). The rates were highest in the SCZ, in spring and winter, and among group-housed calves. These results should provide insights that could lead to better management practices improvements that will help to sustain not only China's dairy farms but also dairy farms operated in the similar climate zones around the world.

A multidisciplinary approach to reduce and refine antimicrobial drugs use for diarrhoea in dairy calves

This study evaluated the impact of a multidisciplinary intervention to reduce and refine antimicrobial drug use for treatment of diarrhoea in dairy calves. The intervention consisted of modification of management practices for disease prevention, health evaluation training of farm staff, implementation of an algorithm directed at antimicrobial therapy for diarrhoeic calves, and monthly farm visits. A combination of retrospective (before intervention period [BP], 1 year) and prospective (immediately after intervention period [AP], 1 year) cohorts were used. Health outcomes measured included incidence of diarrhoea and overall mortality. Antimicrobial treatment rates for diarrhoea and total mass of antimicrobial drugs used at the calf and farm level were also evaluated. Outcomes were assessed using the χ2 or Fisher’s exact test, and odds ratios and 95% confidence intervals (95% CI) were calculated.Records of 2049 and 2251 calves from 10 farms were available in the BP and AP, respectively. Reduced antimicrobial treatment rates occurred on seven farms (P < 0.01), but not on three farms in the AP. A total of 85% (1303/1573) and 18% (310/1698) of diarrhoeic calves were treated with antimicrobial drugs in the BP and AP (P < 0.001), respectively. There were no differences in the incidence of diarrhoea or mortality between periods. In the AP, the use sulfamethazine and trimethoprim decreased on eight farms, one farm discontinued use of lincomycin and spectinomycin, while two farms discontinued use and one reduced use of cephalosporins. This multidisciplinary approach was effective in reducing antimicrobial drug use for calf diarrhoea on dairy farms without negative impacts on calf health.

Using Oral Rehydration Therapy (ORT) in the Community.

For ORT to have a maximum impact on public health it should be used in the community, in the home. A number of programs have been developed over the years to extend ORT to home use. One of the most successful approaches was the Oral Therapy Education Program (OTEP) developed by BRAC, the world’s largest NGO. Mothers were taught in the home by an OTEP worker using seven simple messages and a demonstration. The program, which led to high levels of use and knowledge retention, is described. What the OTEP and other successful home-based programs have demonstrated is that home care of diarrhea using ORS can be effectively implemented and can have a positive impact on the reduction of diarrhea morbidity and mortality.

Shigella-specific antibodies in the first year of life among Zambian infants: A longitudinal cohort study.

IntroductionShigellosis, is a leading cause of moderate-to-severe diarrhoea and related mortality in young children in low and middle income countries (LMICs). Knowledge on naturally acquired immunity can support the development of Shigella candidate vaccines mostly needed in LMICs. We aimed to quantify Shigella-specific antibodies of maternal origin and those naturally acquired in Zambian infants.MethodsPlasma samples collected from infants at age 6, 14 and 52-weeks were tested for Shigella (S. sonnei and S. flexneri 2a) lipopolysaccharide (LPS) antigen specific immunoglobulin G (IgG) and A (IgA) by enzyme-linked immunosorbent assay.ResultsAt 6 weeks infant age, the IgG geometric mean titres (GMT) against S. sonnei (N = 159) and S. flexneri 2a (N = 135) LPS were 311 (95% CI 259–372) and 446 (95% CI 343–580) respectively. By 14 weeks, a decline in IgG GMT was observed for both S. sonnei to 104 (95% CI 88–124), and S. flexneri 2a to 183 (95% CI 147–230). Both S. sonnei and S. flexneri 2a specific IgG GMT continued to decrease by 52 weeks infant age when compared to 6 weeks. In 27% and 8% of infants a significant rise in titre (4 fold and greater) against S. flexneri 2a and S. sonnei LPS, respectively, was detected between the ages of 14 and 52 weeks.IgA levels against both species LPS were very low at 6 and 14 weeks and raised significantly against S. flexneri 2a and S. sonnei LPS in 29% and 10% of the infants, respectively.ConclusionIn our setting, transplacental IgG anti-Shigella LPS is present at high levels in early infancy, and begins to decrease by age 14 weeks. Our results are consistent with early exposure to Shigella and indicate naturally acquired IgG and IgA antibodies to S. flexneri 2a and S. sonnei LPS in part of infants between 14 and 52 weeks of age. These results suggest that a potential timing of vaccination would be after 14 and before 52 weeks of age to ensure early infant protection against shigellosis.

Effects of Dietary Yucca Schidigera Extract and Oral Candida utilis on Growth Performance and Intestinal Health of Weaned Piglets.

Weaning piglets experienced the transformation from breast milk to solid feed and present the proliferation of pathogens, the presence of diarrhea, poor growth performance and even death. Plant extracts and probiotics have certain potential in improving animal growth performance, antioxidant capacity and immune function. The purpose of this study was to explore the effects of dietary yucca schidigera extract (YSE) and oral Candida utilis (CU) on growth performance and intestinal health weaned piglets. According to a 2 × 2 factorial design with the main factors being CU (orally administered 1 mL of 0.85% saline with or without CU; fed basal diet with or without 120 mg/kg YSE), forty 28 d healthy weaned piglets were randomly allocated into four groups of 10 barrows each: (1) piglets fed basal diet and orally administered 1 mL of 0.85% saline (CON); (2) piglets fed basal diet and orally administered 1 mL 1 × 109 cfu/mL C. utilis in 0.85% saline (CU); (3) piglets fed the basal diet containing YSE (120 mg/kg) and orally administered 1 mL of 0.85% saline (YSE); (4) Piglets fed the basal diet containing 120 mg/kg YSE and 1 mL 1 × 109 cfu/mL C. utilis in 0.85% saline (YSE+CU). This study lasted 28 days and evaluated the effects of dietary YSE and oral CU on growth performance, immunity, antioxidant function, ileal morphology, and intestinal microflora in weaned piglets. Dietary YSE increased ADG, the spleen and lymph node indexes, serum GLU, BUN, T-SOD, T-AOC, CAT concentrations, ileal villus height and villus height/crypt depth, jejunal occludin, and β-definsin-2 concentrations and ileal occludin concentration in weaned piglets (P < 0.05); decreased the diarrhea rate and mortality, rectal pH and urine pH, the BUN and MDA concentrations, crypt depth (P < 0.05); improved the diversity of cecal microflora. Orally CU increased ADG, and ADFI, the T-SOD, T-AOC, and CAT activity, ileal villus height, villus height/crypt depth, jejunum occludin, and β-definsin-2 concentrations (P < 0.05); reduced the diarrhea rate and mortality, urine pH, the BUN and MDA concentrations, crypt depth (P < 0.05); improved the diversity of cecal microflora. Dietary YSE and orally CU increased the T-SOD, T-AOC, and CAT activity, villus height/crypt depth, jejunal occludin concentration; reduced the diarrhea rate of weaned piglets by 28%, gastric pH, ileal pH, cecal pH and urine pH, MDA, crypt depth; improved the diversity of cecal microflora. YSE and CU could improve the growth performance, reduce the diarrhea rate, improve intestinal health, and increase the diversity and abundance of cecal microflora in weaned piglets and expected to be used as antibiotics alternative feed additives in the production of weaned piglets.

Isolation and Identification of a Recombinant Porcine Epidemic Diarrhea Virus With a Novel Insertion in S1 Domain.

Porcine epidemic diarrhea virus (PEDV) is the major pathogen that causes diarrhea and high mortality in newborn piglets with devastating impact to the pig industry. Recombination and mutation are the main driving forces of viral evolution and genetic diversity of PEDV. In 2016, an outbreak of diarrhea in piglets occurred in an intensive pig farm in Central China. A novel PEDV isolate (called HNAY) was successfully isolated from clinical samples. Sequence analysis and alignment showed that HNAY possessed 21-nucleotide (nt) insertion in its S1 gene, which has never been reported in other PEDV isolates. Moreover, the sequence of the insertion was identical with the sequence fragment in PEDV N gene. Notably, the HNAY strain exhibited two unique mutations (T500A and L521Y) in the neutralizing epitopes of the S1 protein that were different from those of other PEDV variant strains and CV777-based vaccine strains. Additionally, PEDV HNAY might be derived from a natural recombination between two Chinese variant PEDV strains. Animal experiments demonstrated that HNAY displayed higher pathogenicity compared with two other clinical isolates. This study lays the foundation for better understanding of the genetic evolution and molecular pathogenesis of PEDV.

Differences in Intestinal Barrier Development between Intrauterine Growth Restricted and Normal Birth Weight Piglets.

Simple SummaryAnimals with intrauterine growth restriction (IUGR) are defined as neonates born at term but with low birth weight and a characteristic shape of the head. A number of structural and functional modifications in the IUGR intestine affecting its digestive and absorptive function and impairing intestinal barrier function have been reported in the past. Far less is known about the immune system in the gut of IUGR pigs. Therefore, the aim of the present study was to evaluate the structures of the immune system of the gut mucosa in IUGR neonates. We found that the immune deficiency in the gut mucosa that results from restricted intrauterine development occurs at postnatal day (PD) 7, but it disappears thereafter within a week. However, all examined IUGR piglets had an increased number of intraepithelial leukocytes in the gut mucosa on PD 14. We have shown that the immune system of the gut of IUGR piglets is able to quickly compensate for the immunological deficiency postnatally and hardly shows any morphological disabilities in later life.Intrauterine growth restricted (IUGR) piglets are born at term but have low birth mass and a characteristic shape of the head. Impaired general condition, especially in intestinal function, leads to an increase in the occurrence of diarrhoea and high mortality in the first days of life. So far, the mechanical and immunological gut barrier functions in IUGR are poorly understood. The aim of this study was to microscopically evaluate the early postnatal changes in the gut mucosa occurring in IUGR piglets. Whole-tissue small intestine samples were collected from littermate pairs (IUGR and normal) on postnatal day (PD) 7, 14 and 180 and analysed by light microscopy. We found that in the IUGR piglets, the percentage of intraepithelial leukocytes was reduced in the duodenum on PD 7, but it increased in the proximal and middle jejunum both on PD 7 and PD 14, which suggested the development of an inflammatory process. The number of goblet cells was also reduced on PD 14. The average size of the Peyer’s patches in the distal jejunum and ileum showed significant reduction on PD 7 as compared to normal pigs; however, on PD 14, it returned to normal. On PD 180, we did not find any differences in the measured parameters between the IUGR and the normal pigs. In conclusion, we found that in one-week-old IUGR pig neonates, the gut barrier and the immune system structures display signs of retarded development but recover within the second postnatal week of life.

Combination of conditional cash transfer program and environmental health interventions reduces child mortality: an ecological study of Brazilian municipalities

BackgroundThis study aims to assess the interactive effects of Brazilian public interventions, environmental health programs (access to water, sanitation and solid waste collection) and a Conditional Cash Transfer Program (PBF), on the mortality reduction due to diarrhea and malnutrition among children under 5 years old.MethodsThe study design is ecological, with longitudinal analysis in a balanced panel. The period covered is 2006 to 2016, including 3467 municipalities from all regions of the country, which resulted in 38,137 observations. The generalized linear models were adjusted considering the Negative Binomial (NB) distribution for the number of deaths due to malnutrition and diarrhea, with fixed effects. NB models with and without zero-inflation were assessed. Subsequent interaction models were applied to assess the combined effects of the two public policies.ResultsIn relation to the decline of mortality rates due to diarrhea in the municipalities, positive effect modification were observed in the presence of: high coverage of the target population by the PBF and access to water, 0.54 (0.28–1.04) / 0.55 (0.29–1.04); high coverage by the total population by the PBF and access to water, 0.97 (0.95–1.00) and high coverage by the total population by the PBF and access to sanitation, 0.98 (0.97–1.00). Decline on diarrhea mortality was also observed in the joint presence of high coverage of solid waste collection and access to water, categories 1 (> 60% ≤85%): 0.98 (0.96–1.00), 0.98 (0.97–1, 00) and 2 (> 85% ≤ 100%): 0.97 (0.95–0.98), 0.97 (0.95–0.99). Negative effect modification were observed for mortality due to malnutrition in the presence of simultaneous high coverage of the total population by the PBF and access to sanitation categories 1 (≥ 20 < 50%): 1.0061 (0.9991–1.0132) and 2 (≥ 50 < 100%): 1.0073 (1.0002–1.0145) and high coverage of the total population by the PBF and solid waste collection, 1.0004 (1.0002–1.0005), resulting in malnutrition mortality rates increase.ConclusionImplementation of environmental health services and the coverage expansion by the PBF may enhance the prevention of early deaths in children under 5 years old due to diarrhea, a poverty related disease.

Molecular Epidemiology of Human Cryptosporidiosis in Low- and Middle-Income Countries.

Cryptosporidiosis is one of the most important causes of moderate to severe diarrhea and diarrhea-related mortality in children under 2 years of age in low- and middle-income countries. In recent decades, genotyping and subtyping tools have been used in epidemiological studies of human cryptosporidiosis. Results of these studies suggest that higher genetic diversity of Cryptosporidium spp. is present in humans in these countries at both species and subtype levels and that anthroponotic transmission plays a major role in human cryptosporidiosis. Cryptosporidium hominis is the most common Cryptosporidium species in humans in almost all the low- and middle-income countries examined, with five subtype families (namely, Ia, Ib, Id, Ie, and If) being commonly found in most regions. In addition, most Cryptosporidium parvum infections in these areas are caused by the anthroponotic IIc subtype family rather than the zoonotic IIa subtype family. There is geographic segregation in Cryptosporidium hominis subtypes, as revealed by multilocus subtyping. Concurrent and sequential infections with different Cryptosporidium species and subtypes are common, as immunity against reinfection and cross protection against different Cryptosporidium species are partial. Differences in clinical presentations have been observed among Cryptosporidium species and C. hominis subtypes. These observations suggest that WASH (water, sanitation, and hygiene)-based interventions should be implemented to prevent and control human cryptosporidiosis in low- and middle-income countries.

Heparan Sulfate Is a Cellular Receptor for Enteric Human Adenoviruses.

Human adenovirus (HAdV)-F40 and -F41 are leading causes of diarrhea and diarrhea-associated mortality in children under the age of five, but the mechanisms by which they infect host cells are poorly understood. HAdVs initiate infection through interactions between the knob domain of the fiber capsid protein and host cell receptors. Unlike most other HAdVs, HAdV-F40 and -F41 possess two different fiber proteins—a long fiber and a short fiber. Whereas the long fiber binds to the Coxsackievirus and adenovirus receptor (CAR), no binding partners have been identified for the short fiber. In this study, we identified heparan sulfate (HS) as an interaction partner for the short fiber of enteric HAdVs. We demonstrate that exposure to acidic pH, which mimics the environment of the stomach, inactivates the interaction of enteric adenovirus with CAR. However, the short fiber:HS interaction is resistant to and even enhanced by acidic pH, which allows attachment to host cells. Our results suggest a switch in receptor usage of enteric HAdVs after exposure to acidic pH and add to the understanding of the function of the short fibers. These results may also be useful for antiviral drug development and the utilization of enteric HAdVs for clinical applications such as vaccine development.

Water purification efforts and the black‐white infant mortality gap, 1906–1938

According to Troesken (2004), efforts to purify municipal water supplies at the turn of the 20th century dramatically improved the relative health of Blacks. There is, however, little empirical evidence to support the Troesken hypothesis. Using city-level data published by the U.S. Bureau of the Census for the period 1906–1938, we explore the relationship between water purification efforts and the Black-White infant mortality gap. Our results suggest that, while water filtration was effective across the board, adding chlorine to the water supply reduced mortality only among Black infants. Specifically, chlorination is associated with a 9 percent reduction in Black infant mortality and a 10 percent reduction in the Black-White infant mortality gap. We also find that chlorination led to a substantial reduction in the Black-White diarrhea mortality gap among children under the age of two, although this estimate is measured with less precision.

A STUDY OF CLINICAL AND SUBCLINICAL CHARACTERISTICS IN ACUTE DIARRHEA IN CHILDREN FROM 2 MONTHS TO 5 YEARS IN HUE CENTRAL HOSPITAL

Background: Mortality of diarrhea have decreased significantly starting in 2000, from about 1.2 million deaths among children under 5 years old to 526,000 in 2015 - a 57% reduction. Despite this, diarrhea remains the second most common cause of death among children under 5, closely following pneumonia. Objectives: To describe clinical and subclinical characteristics of acute diarrhea in children from 2 months to 5 years of age. Subjects and methods: 148 patients from 2 months to 5 years of age who were diagnosed with acute diarrhea for treatment in General Pediatrics Department 1 - Pediatric Center - Hue Central Hospital from April 2018 to April 2019. Set up a cross-sectional study. Results: The group of 2 - 24 months of age accounted for the highest proportion (83.1%), no dehydration (84.5%), dehydration (15.5%), no severe dehydration. The common electrolyte disturbance is hyponatremia (15.5%) and hypokalemia (32.4%). Conclusion: Acute diarrhea in children usually lasts less than 7 days; The common symptoms are vomiting, fever, inflammation of the upper respiratory tract; Laboratory tests showed that peripheral leukocytes were usually not increased and most of them were caused by Rotavirus. Key words: clinical, subclinical, acute diarrhea

Effects of Taraxacum and Astragalus extracts combined with probiotic Bacillus subtilis and Lactobacillus on Escherichia coli–infected broiler chickens

Diarrhea caused by Escherichia coli (E. coli) is one of the most common diseases that affects the growth and development of poultry. This study was conducted to investigate the synergistic effects of traditional Chinese medicine (TCM) combined with probiotics against E. coli infection and its mechanism in broiler chickens. The optimal proportion formula TCM and probiotics was screened by orthogonal test and range analysis method; the in vitro antibacterial activity was based on the Oxford cup method. Isolated pathogenic E. coli was injected subcutaneously into the neck of the broilers to establish an E. coli–infected model. The broilers were administrated with drugs in drinking water daily for 7 d before and after E. coli infection. The diarrhea rate, mortality, body weight (BW) gain, feed intake, immune organ index, intestinal and hepatic histopathological changes were monitored. The expression of IL-2, IL-10, and TLR-4 mRNA in the intestinal tissues was measured by RT-PCR. Our results showed that the optimal proportion formula of Taraxacum extracts: total flavonoids of Astragalus: polysaccharides of Astragalus: probiotics was 5: 2: 2: 2; TCM combined with probiotics was highly sensitive to E. coli. TCM combined with probiotics synergistically increased BW gain, decreased the diarrhea rate and mortality of broilers, alleviated intestinal and hepatic pathological changes, accompanied by the increase of IL-2 and IL-10 mRNA expression and the inhibition of TLR-4 mRNA expression. It suggests that the combination of TCM and probiotics may produce a synergistic protective effect against E. coli infection by improving the indicators of diarrhea and regulating the expression of IL-2, IL-10, and TLR-4 mRNA in broiler chickens. The synergistic interactions between TCM and probiotics represent a promising strategy for the treatment of E. coli infection.

Pathological diagnosis of infectious bursal disease in 24 weeks old vaccinated commercial laying hens in Kagarko, Nigeria: a case report

Infectious bursal disease (Gumboro disease) was diagnosed in 24 weeks old vaccinated commercial laying hens. Clinical signs observed were severe depression, greenish-white diarrhea and high mortality. Postmortem findings observed were ruffled feathers, congestion of the lungs, enlarged kidneys, enlarged spleens, congested ovarian follicles, pale livers, haemorrhages of the mucosa of the proventriculus and ventricular junction, enlarged, oedematous and haemorrhagic bursa of Fabricius and haemorrhages of caecal tonsils. Histopathology findings showed severe congestion of kidneys, presence of red blood cells in the interstitial spaces, tubular degenerative necrosis with dissolution of nuclear material, glomerular atrophy and increase in capsular spaces. There was severe follicular inflammation of the bursa of Fabricius evident by massive presence of inflammatory cells. Infectious bursal disease was diagnosed based on the gross pathological and histopathological findings. King herbs was administered for 5 days at dose rate 2ml/L in the birds drinking water ad-lib and the client was advised to isolate and treat the sick birds with the drug and to enforce strict biosecurity measures. There was no mortality after the treatments.

Immunogenicity and duration of immunity of the polyvalent vaccine against chicken salmonellosis

Poultry salmonellosis causes serious economic damage and must be prevented by disinfection, zoohygienic measures or by vaccination. To develop a new polyvalent vaccine against poultry salmonellosis, we used bacterial strains of Salmonella enteritidis, S. typhimurium and S. gallinarum. Antigenic and immunogenic efficacy of the vaccine was tested on specific-pathogen free chickens, which were divided into five groups of 10 birds in each group and were vaccinated intramuscularly at 8 and 12 weeks: group A (non-immunized control), group B (S. enteritidis mono-vaccine), group C (S. typhimurium mono-vaccine), group D (S. gallinarum mono-vaccine) and group E (trivalent vaccine Polimun Salmo). None of the immunized birds showed such adverse reactions as abnormal behaviour, mortality or signs of anorexia, depression or diarrhea. Two weeks after the revaccination, 5 birds in each group were challenged by watering 3 cm3 of working suspensions of S. gallinarum, S. typhimurium and S. enteritidis control strains at a concentration of 1 × 109 CFU. 72 h after the challenge, faeces were collected from all chickens in each group to identify Salmonella excretion with faeces, and the chickens were euthanized. Significant protection against the virulent challenge was observed in all immunized groups based on mortality and post-mortem lesions compared with the non-immunized control group. Blood samples were selected weekly from 5 chickens of each group for 184 days. The antigenic efficacy of the vaccines was studied by reaction of haemagglutination in the obtained serum. The potent antigen-specific response to lymphocyte activation found in all immunized groups indicated the induction of immune responses. Overall, the results showed that persistent immunity is formed in 4 weeks after the revaccination and lasts for a productive period. Immune response of chickens on day 184 after vaccination with Polimun Salmo was 1: 647, indicating that the developed polyvalent vaccine against common serovars of S. enterica in poultry is effective and immunogenic and can be further used in field studies.

Evolution of Transmissible Gastroenteritis Virus (TGEV): A Codon Usage Perspective.

Transmissible gastroenteritis virus (TGEV) is a coronavirus associated with diarrhea and high mortality in piglets. To gain insight into the evolution and adaptation of TGEV, a comprehensive analysis of phylogeny and codon usage bias was performed. The phylogenetic analyses of maximum likelihood and Bayesian inference displayed two distinct genotypes: genotypes I and II, and genotype I was classified into subtypes Ia and Ib. The compositional properties revealed that the coding sequence contained a higher number of A/U nucleotides than G/C nucleotides, and that the synonymous codon third position was A/U-enriched. The principal component analysis based on the values of relative synonymous codon usage (RSCU) showed the genotype-specific codon usage patterns. The effective number of codons (ENC) indicated moderate codon usage bias in the TGEV genome. Dinucleotide analysis showed that CpA and UpG were over-represented and CpG was under-represented in the coding sequence of the TGEV genome. The analyses of Parity Rule 2 plot, ENC-plot, and neutrality plot displayed that natural selection was the dominant evolutionary driving force in shaping codon usage preference in genotypes Ia and II. In addition, natural selection played a major role, while mutation pressure had a minor role in driving the codon usage bias in genotype Ib. The codon adaptation index (CAI), relative codon deoptimization index (RCDI), and similarity index (SiD) analyses suggested that genotype I might be more adaptive to pigs than genotype II. Current findings contribute to understanding the evolution and adaptation of TGEV.

Subcellular localization of the porcine deltacoronavirus nucleocapsid protein.

Porcine deltacoronavirus (PDCoV) has been recently identified as an emerging enteropathogenic coronavirus that mainly infects newborn piglets and causes enteritis, diarrhea and high mortality. Although coronavirus N proteins have multifarious activities, the subcellular localization of the PDCoV N protein is still unknown. Here, we produced mouse monoclonal antibodies against the PDCoV N protein. Experiments using anti-haemagglutinin antibodies and these monoclonal antibodies revealed that the PDCoV N protein is shuttled into the nucleolus in both ectopic PDCoV N-expressing cells and PDCoV-infected cells. The results of deletion mutagenesis experiments demonstrated that the predicted nucleolar localization signal at amino acids 295–318 is critical for nucleolar localization. Cumulatively, our study yielded a monoclonal antibody against the PDCoV N protein and revealed a mechanism by which the PDCoV N protein translocated into the nucleolus. The tolls and findings from this work will facilitate further investigations on the functions of the PDCoV N protein.

Aloe extract inhibits porcine epidemic diarrhea virus in vitro and in vivo

Porcine epidemic diarrhea virus (PEDV) causes severe diarrhoea and high mortality in neonatal suckling piglets, leading to significant economic losses to the swine industry. Currently there are no adequate control strategies against circulating PEDV variants, making an urgent need to exploit effect antiviral therapies to compensate for vaccines. Here, we report that Aloe extract can hamper completely the proliferation of PEDV at a non-cytotoxic concentration of 16 mg/mL determined by CCK-8 assay in Vero and IPEC-J2 cells in vitro. Furthermore, time course analysis indicated the extract exerted its inhibition at the late stage of the viral life cycle. Moreover, we also confirmed that the extract can inactivated PEDV directly but did not act on the viral genome and S1 protein. Importantly, the extract at a relatively safety concentration of 100 mg/kg of body weight, which was confirmed in mice, could reduce virus load and pathological change in intestinal tract of pigs and protect newborn piglets from lethal challenge with highly pathogenic PEDV variant GDS01 infection, indicating that Aloe extract efficiently inhibited PEDV infection in vivo. Collectively, our findings suggest that the aqueous extract from the Aloe could inhibit PEDV replication in vitro and in vivo and might be a good target for drug development against PEDV.