Weak Precipitin Ring from A Fecal Specimen with Markedly Elevated Alpha-1 Antitrypsin Level Measured by Radial Immunodiffusion

Radial immunodiffusion (RID) is a classic methodology for antigen quantification that relies on the development of a distinct precipitin ring from precipitated antigen-antibody complex. As the precipitin ring grows, the precipitate at the inner edge of the ring constantly dissolves due to excess antigen flooding from the point of application, while new precipitate forms at the leading edge of the ring. RID plates with anti-human alpha-1-antitrypsin are prepared in our lab to measure fecal alpha-1 antitrypsin (A1AT) to aid in the diagnosis and monitoring of Protein Losing Enteropathy (PLE). The procedure has routinely produced precipitin rings with small radii and distinct edges after incubating at room temperature for 48 hours. Unexpectedly, a fecal specimen from a patient produced an extremely weak and large precipitin ring that could have been easily overlooked. Dilution studies confirmed a highly elevated A1AT level of 67 mg/g dry stool. The very weak and large precipitin ring was reproduced with a spiked specimen with similar A1AT concentration and kept expanding for several days until a distinct ring was formed eventually. Our data highlights a rare example of high-dose hook effect in RID and calls for meticulous attention and caution when reading and interpreting gel-based immunoassays with unexpected markedly elevated results to avoid additional confirmatory testing. In these cases, we recommend repeat testing with diluted specimens.

Pragmatic Strategy for Fecal Specimen Storage and the Corresponding Test Methods for Clostridioides difficile Diagnosis

The quality of fecal specimens is one of the factors responsible for successful Clostridioides difficile infection (CDI) diagnosis. The quality depends largely on the storage conditions, including the temperature and time period. In this study, we organized the outputs of previous studies, filled experimental gaps in the knowledge of storage conditions, and introduced a pragmatic strategy for fecal storage for CDI diagnosis. A 5-step pathway was adopted to develop the fecal specimen storage strategy as follows: step 1, bibliomic analysis; step 2, experimental gap-filling; step 3, comparative evaluation; step 4, strategy development; step 5, internal review. Step 1 identified eight articles providing experimental information on the effects of fecal specimen storage conditions on the effectiveness of C. difficile detection methods. Step 2 provided additional quantitative data on C. difficile vegetative and spore cell viability and DNA stability. All previous and current results were compared (step 3). In step 4, fir general and nine special strategies were developed, followed by an internal review of the overall approaches (step 5). It is recommended to separate fecal samples into aliquots before testing and storing them. It is particularly recommended that fecal specimen samples be stored for CDI diagnosis at 4 °C for up to 60 days for all test methods.

Prevalence of Intestinal Helminthes with Respect to Age, Sex and Breeds of Chicken Slaughtered at Eke Awka Market, Awka, Anambra State, Nigeria

Aim: This research was designed to assess the prevalence of intestinal helminthes with respect to age, sex and breeds of chicken slaughtered at Eke-Awka Market, Awka, Anambra State, Nigeria. Methodology: Fecal specimen was obtained from 150 Chicken slaughtered at Eke-Awka, Market, Anambra State, Nigeria from August to October 2019. The Fecal specimen was processed following floatation and the formalin-ethyl acetate concentration techniques. The collected data was analyzed using descriptive statistics to get the percentage prevalence and chi-square to determine the association of prevalence in relation to age, sex and breeds of the chickens; level of significance was set at P < 0.05. Results: Out of the 150 fecal specimens examined, 63 were found to be infected with intestinal helminthes parasites, giving a prevalence of 42%. The parasites found were Ascaridia galli, Heterakis gallinarum and Raillietina cesticillus. Raillietina cesticillus was observed to have the highest prevalence which is 16.7%. With respect to gender, a total of 38 males were found to be infected by helminthes parasites and the prevalence among males was 25.3%. Raillietina cesticillus infected the highest number of male chickens 17 (44.7%). A total of 25 female chickens were infected, with a prevalence of 16.7%, Ascaridia galli infected the highest number of female chickens 11 (44%). With respect to age groups, a total of 24 chickens aged (0 – 5) months were infected with a prevalence of 16%. Raillietina cesticillus had the highest infection rate 12 (50%). In the age group (6 – 10) months, a total of 21 chickens were infected giving a prevalence rate of 14%. Ascaridia galli recorded the highest infection rate 9 (49.9%). Among age group 11 months and above, the total number of infected chickens was 18, giving a prevalence rate of 12%. Heterakis gallinarum recorded the highest infection rate 8 (44.4%). With respect to breeds, a total of 28 old layer chickens were found to be infected by helminthes parasites and its prevalence was 18.7%, Raillietina cesticillus had the highest infection rate 13 (46.4%). The broiler had a total infection rate of 22 which gave a prevalence of 14.7%. Raillietina cesticillus had the highest infection rate of 9 (40.9%). Among the Native chickens, the total number of infections was 13 and its prevalence rate was 8.7%. Ascaridia galli recorded the highest infection rate which was 8 (61.5%). Conclusions: The overall prevalence of intestinal helminthes was 42%, the helminthes parasites found were Ascaridia galli, Heterakis gallinarum and Raillietina cesticillus. Raillietina cesticillus was observed to have the highest prevalence 16.7%. Government and poultry owners should ensure that good caging and management systems are adopted to prevent the spread of intestinal helminthes among chickens so as to ensure maximum output from poultry production.

Effects of probiotic on gut microbiota in children with acute diarrhea: a pilot study

Background Acute diarrhea is a common health problem in Indonesia. During acute diarrhea, changes in gut microbiota are marked by decrease beneficial microbes Bifidobacterium and Lactobacillus, and increased pathogenic bacteria Enterobacter and Clostridium. Such microbial imbalances are known as dysbiosis. Treatment with probiotics may help repair dysbiosis, quicken healing time, and decrease complications. Objective To assess for dysbiosis during acute diarrhea, and determine if it can be normalized by probiotic treatment. Methods This placebo-controlled, unblinded clinical trial was performed in Budhi Asih District Hospital, Jakarta, from January to March 2018. Twenty-four children age 6-24 months with acute diarrhea and 12 healthy children were enrolled. First fecal specimen was collected for all subjects and analyzed using non-culture real time PCR to count the population of Lactobacillus, Bifidobacterium, Enterobacter, Clostridium, and all bacteria. Children with diarrhea were assigned to probiotic or placebo treatment for 5 days and the second fecal specimen was analyzed two weeks after the diarrhea subsided. Results Prior to treatment, significant higher amounts of Lactobacillus were observed in children with acute diarrhea than in healthy controls [median (interquartile range/IR): 1.52x103 (1.22x104) vs. 6.87x10 (2.41x102), respectively; proportion in percentage (from total bacteria population): 0.044% vs. 0.003%, respectively]. However, median (IR) Clostridium was significantly higher in healthy controls than in children with acute diarrhea [2.37x102 (4.64x103) vs. 4.67 (1.50x102), respectively (P<0.05), with proportion of 0.01% vs. 0.0001%, respectively]. Children who received probiotics had significantly higher count of Bifidobacterium compared to the placebo group [1.94x104 (4.97x104) vs. 1.74x103 (2.08x107), respectively, with proportion of 0.394% vs. 0.081%, respectively]. Conclusion This pilot study do not find evidence of dysbiosis in children with acute diarrhea. Group who received probiotic had higher Bifidobacterium count compared towards those who received placebo.

Effects of probiotic on gut microbiota in children with acute diarrhea: a pilot study

Background Acute diarrhea is a common health problem in Indonesia. During acute diarrhea, changes in gut microbiota are marked by decrease beneficial microbes Bifidobacterium and Lactobacillus, and increased pathogenic bacteria Enterobacter and Clostridium. Such microbial imbalances are known as dysbiosis. Treatment with probiotics may help repair dysbiosis, quicken healing time, and decrease complications. Objective To assess for dysbiosis during acute diarrhea, and determine if it can be normalized by probiotic treatment. Methods This placebo-controlled, unblinded clinical trial was performed in Budhi Asih District Hospital, Jakarta, from January to March 2018. Twenty-four children age 6-24 months with acute diarrhea and 12 healthy children were enrolled. First fecal specimen was collected for all subjects and analyzed using non-culture real time PCR to count the population of Lactobacillus, Bifidobacterium, Enterobacter, Clostridium, and all bacteria. Children with diarrhea were assigned to probiotic or placebo treatment for 5 days and the second fecal specimen was analyzed two weeks after the diarrhea subsided. Results Prior to treatment, significant higher amounts of Lactobacillus were observed in children with acute diarrhea than in healthy controls [median (interquartile range/IR): 1.52x103 (1.22x104) vs. 6.87x10 (2.41x102), respectively; proportion in percentage (from total bacteria population): 0.044% vs. 0.003%, respectively]. However, median (IR) Clostridium was significantly higher in healthy controls than in children with acute diarrhea [2.37x102 (4.64x103) vs. 4.67 (1.50x102), respectively (P<0.05), with proportion of 0.01% vs. 0.0001%, respectively]. Children who received probiotics had significantly higher count of Bifidobacterium compared to the placebo group [1.94x104 (4.97x104) vs. 1.74x103 (2.08x107), respectively, with proportion of 0.394% vs. 0.081%, respectively]. Conclusion This pilot study do not find evidence of dysbiosis in children with acute diarrhea. Group who received probiotic had higher Bifidobacterium count compared towards those who received placebo.

Parasitosis of Ruminant Animal in Private Farms of Vysokogorny and Laishevskiy Districts of the Republic of Tatarstan

The purpose of the research is to study the species composition of intestinal parasitosis agents in ruminant animals in citizens’ private farms in Vysokogorny and Laishevskiy districts of the Republic of Tatarstan. Materials and methods. The work has been executed in the Epizootology, Parasitology and Radiobiology Department at the Kazan State Academy of Veterinary Science named after N.E.Bauman and in citizens’ private farms in Vysokogorny and Laishevskiy districts of the Republic of Tatarstan during autumn-winter period in 2016-2017. 586 fecal specimen, including 364 samples of cattle, 168 samples of sheep and 54 samples of goats, have been studied by the modified method of flotation. Animals extent of invasion were determined as well as average number of helminth eggs and eimeria oocyst in 1 g of feces were calculated with the help of VIGIS count chamber. Kheisin key was used to identify eimeria types. Statistical analysis of numeric material was carried out with the help of Microsoft Excel spreadsheet processor. Results and discussion. Eggs of helminth classes Fasciola, Moniezia, Nematodirus, Trichocephalus as well as eimeria oocyst had been founded in cattle and small ruminants fecal specimen. On investigated areas the degree of infection by fascioles was 28.2%, by moniezia 41.5%, by nematodirus 52.7%, by trichocephalus 28.3%, and by eimeria 51.3%. Small ruminants were infected with fascioles in 56.5%, moniezia in 52.7%, thysaniezia in 15.7%, nematodirus in 58.1%, trichocephalus in 38.5%, and eimeria in 21.3%. Taking into consideration prevalence of parasitosis of ruminant animal in private farms of Vysokogorny and Laishevskiy districts of the Republic of Tatarstan it is necessary to make up a plan of antiparasitic measures inclusive of climate pattern and developmental biology of agents.

Spread of eimeriosis among calves under the conditions of south European part of Russia

The purpose of the research: to define the degree of spread of eimeriosis among calves under the conditions of south European part of Russia. Materials and methods. The researches were carried out in 11 entities of Krasnodar Territory, Stavropol Territory and Rostov Region in 2016-2017. The material for the researches was fecal specimen of cattle from entities of different forms of ownership. Total of 975 fecal specimen of cattle were studied. Fecal specimens were studied according to Fulleborn’s method. Invasion extensiveness (IE, %) and average number of oocysts in 1 g of feces were determined. Seasonal dynamics of infection of young stock cattle was studied by studies of fecal specimens of 38 heads in January - February, 43 heads in March - May, 64 heads during summer and 55 heads during autumn period. Influence of the stock density of calves in a pen on infection by eimeria was studied on 249 calves, among which 62 animals were kept in individual pens, 25 animals were kept in a pen for 2-5 animals, 28 calves were kept in a pen for 6-10 animals, 55 calves were kept in a pen for 11-15 animals and 75 calves were kept in a pen for 16 animals and more. Fecal specimens were studied three times. Obtained results were processed statistically. Results and discussion. Animals were infected by eimeria regardless of the type of entity. Average infection of the cattle in all tested entities was 47.43% when 180.5±8.3 samples of oocysts were found in average in 1 g of feces. In entities the infection of young stock cattle varied from 0 to 100% with maximum result at Urozhaynoe CJSC of Stavropol Territory when 362.3±9.7 samples of oocysts were found in 1 g of feces. To the fullest extent calves were infected by eimeria at the age of 3-4 months (IE 70.73%) when 364.3±8.2 samples of oocysts were found in 1 g of feces in average. Mature animals were infected minimally (IE 20.0%). Maximum animal infection was in spring and in autumn, 53.49% and 61.81% respectively when 282.3±7.5 and 324.6±8.3 samples of oocysts were available in 1 g of feces. It was established, that while rising the stocking rate of calves in the pen the eimeria infection also greatly increases. Minimal invasion was in calves which were kept in individual pens.