Dynamics of indicators of cellular immunity in conditions of acute generalized peritonitis in rats

The course of peritonitis depends on the state of the immune system and the adequacy of the immune response. The aim of this investigation was to explore the features of the cellular immune response cell in rats with simulated acute generalized peritonitis (AGP). The study was conducted on 32 rats, divided into two groups: the main group – 24 animals with simulated peritonitis; control– 8 intact animals. In the animals of the main group AGP were modeleted by injecting of 10 % filtered stool suspension into the abdominal cavity of the study rats at a dose of 0.5 ml per 100 g of body weight. Removal of material for histological examination was carried out on days 1, 3 and 7. Indicators of cellular immunity were determined by a method that was based on the interaction of fluorescently labeled monoclonal antibodies with lymphocyte surface antigens. In the main group of animals, all indicators of the cellular immunity gradually decreased from 1 to 7 days of the experiment. The concentration of CD3+ - cells decreased by 1.90 times per day, by 1.97 times by 3 days, and by 2.10 times by 7 days, compared with intact animals. Suppression of the cellular immunity was observed after modeling of AGP, which is combined with a decrease in the number of both CD3+ lymphocytes and the main subpopulations of CD4+, CD8+ and CD16+ cells. The greatest decrease in the cellular level of immunity was observed on day 7 of the experiment.

Comparison of algorithms for the detection of enteroviruses in stool specimens from children diagnosed with Acute Flaccid Paralysis

ABSTRACTWith poliovirus eradication within reach, the WHO has included in its recommendations a cell-culture independent algorithm for enterovirus surveillance. This study was designed to compare both the cell culture dependent and independent algorithms and assess how either might impact our perception of the diversity of enterovirus types present in a sample.Sixteen paired samples (16 isolates from RD cell culture and their corresponding stool suspension. i.e. 32 samples) from AFP cases in Nigeria were analyzed in this study. One of these 16 sample pairs (the control) was previously identified and confirmed as poliovirus 2 (PV-2). All the samples were subjected to RNA extraction, cDNA synthesis, RT-snPCR (the WHO recommended cell-culture independent algorithm) and its modifications for co-infection detection and resolution. Amplicons were sequenced and strains identified using the enterovirus genotyping tool and phylogenetic analysis.The enterovirus diversity was shown to be the same between RD cell culture isolates and fecal suspension for the control and five (7, 10, 11, 12 & 14) of the samples analyzed. It was however, different for the remaining 10 (62.5%) samples analyzed. Fourteen different enterovirus types were identified in this study. To be precise, 9 (CV-B4, E6, E7, E13, E14, E19, E29, EV-B75 and EV-B77) and 5 (CV-A1, CV-A11, CV-A13, EV-C99 and PV2) EV-B and EV-C types, respectively where detected in this study. It is crucial to mention that E19 and EV-B75were only recovered from RD cell culture isolates while E14, EV-B77, CV-A11 and CV-A13 were only recovered from fecal suspension.The results of this study show that both the cell culture dependent and independent protocols recommended by the WHO for enterovirus detection unavoidably bias our perception of the diversity of enterovirus types present in a sample. Hence, rather than jettison one for the other, effort should be directed at harmonizing both for increased sensitivity.

Comparison of Algorithms for the Detection of Enteroviruses in Stool Specimens from Children Diagnosed with Acute Flaccid Paralysis

This study was designed to compare both the cell culture dependent and independent enterovirus detection algorithms recommended by the WHO and assess how either might impact our perception of the diversity of enterovirus types present in a sample. Sixteen paired samples (16 isolates from RD cell culture and their corresponding stool suspension, i.e., 32 samples) from AFP cases in Nigeria were analyzed in this study. All the samples were subjected to RNA extraction, cDNA synthesis, the WHO recommended RT-snPCR, and its modification. Amplicons were sequenced and strains identified. Enterovirus diversity was the same between the isolates and fecal suspension for the control and five of the samples. It was, however, different for the remaining 10 (62.5%) samples. Nine (CV-B4, E6, E7, E13, E14, E19, E29, EV-B75, and EV-B77) and five (CV-A1, CV-A11, CV-A13, EV-C99, and PV2) EV-B and EV-C types, respectively, were detected. Particularly, E19 and EV-B75 were only recovered from the isolates while E14, EV-B77, CV-A11, and CV-A13 were only recovered from fecal suspension. Both the cell culture dependent and independent protocols bias our perception of the diversity of enterovirus types present in a sample. Hence, effort should be directed at harmonizing both for increased sensitivity.

IDENTIFIKASI KRIPTOSPORIDIOSIS DI PASIEN ANAK HIV DENGAN DIARE KRONIS DI RUANG GASTRO ANAK

Based on the results of overseas researchers, Cryptosporidiosis occurs in immunosuppressive cases with chronic diarrhoea. In this study the researchers would like to know exactly whether that Cryptosporidiosis occurs also in paediatric HIV patients. The latest data show that the incidence of opportunistic infection is characterized by persistent diarrhoea and severe malnutrition as a complication of the paediatric HIV-infected patients is increasing. The objects of the research were fifteen paediatric HIV-infected patients which treated at the Paediatric Gastro Ward of Dr. Soetomo Hospital Surabaya due to persistent diarrhoea. Paediatric patients were less than five years old, suffered persistent diarrhoea more than two weeks with severe malnutrition. Stool specimens were transported using 10% formalin. The stool suspension was filtered, and distilled water was added followed by centrifugation (sedimentation method). The precipitate material was placed on a glass object and dried, and then fixed by methanol and stained with Acid Fast Staining and trichrome staining. The protozoa Cryptosporidium spp. was observed under a binocular microscope with 100× magnification (immersion oil) objective. The result was confirmed as positive if a red spherical or oval formation of oocyste of 4–6 micron appeared. Sixty percent of the 15 paediatric HIV-infected patients with chronic diarrhoea showed positive cryptosporidiosis. Cryptosporidiosis is one of the opportunistic infections resulting in chronic diarrhoea in paediatric HIV-infected patients. The results of the present research indicate that the enteric parasite Cryptosporidium spp. was the main cause of persistent diarrhoea in paediatric HIV-infected patients

Inactivation and UV Disinfection of Murine Norovirus with TiO2 under Various Environmental Conditions

ABSTRACT We studied inactivation and UV disinfection of murine norovirus (MNV) as a surrogate for human norovirus. We investigated the effects of different surface characteristics, temperatures, and NaCl concentrations on MNV survival using both a plaque assay and a real-time TaqMan reverse transcription (RT)-PCR assay. MNV survived more than 40 days on diaper material, on gauze, and in a stool suspension. Compared to inactivation at lower temperatures (−20 and 4°C), inactivation of MNV was greater at higher temperatures (18 and 30°C). On the surface of both gauze and diaper material, there was a <2-log10 reduction in the amount of infectious MNV in 40 days after incubation at both −20 and 4°C, compared to a >5-log10 reduction after incubation at 30°C in 24 days. MNV survived better in a stool suspension than on the surface of gauze or diaper material. A higher salt concentration increased the rate of inactivation of MNV. In 72 h, <0.3-, 1.5-, and 2.5-log10 reductions in the amount of infectious MNV occurred in distilled water and 0.5 and 1 M NaCl, respectively. We observed only minor reductions in the numbers of viral RNA copies as quantified by real-time TaqMan RT-PCR regardless of the temperature, the salt concentration, or the suspending medium. We also evaluated UV disinfection of infectious MNV with and without TiO2. The amount of MNV was significantly reduced by 254-nm UV with and without TiO2. When 25 mJ/cm2 UV was used, 3.3- and 3.6-log10 reductions in the amounts of infectious MNV occurred with and without TiO2, respectively. Our results demonstrate that MNV can persist in various environmental conditions and can be efficiently controlled by UV disinfection.