Potential Rhizopus spp. in control the growth of Aspergillus flavus FNCC6109 in broiler chicken concentrate feed

Aspergillus flavus contamination of agriculture in Indonesia can cause problems to animal health and productivity. Some factors can support the appearance of contamination in feed, especially temperature and humidity. The main objective of this research was to investigate potency of Rhizopus spp. on inhibit A. flavus FNCC6109 in broiler chicken concentrate feed. The experiments were conducted dual culture method and the inhibition test of the Rhizopus spp. filtrate culture was incubated for 3, 4 and 5 days on in vitro. The in vivo test was directly applied in broiler chicken concentrate feed which added Rhizopus spp. filtrate culture concentration at 10% (v/v), 20% (v/v), 30% (v/v), 40% (v/v), dan 50% (v/v). The results showed that the Rhizopus spp. filtrate culture significantly (P?0,05) to inhibit the growth of A. flavus FNCC6109 both in vitro and in vivo. The percentage inhibition of Rhizopus spp. filtrate culture incubated for 5 days showed 67,47±2,10% relatively better results than 3 and 4 days, and therefore was used in the in vivo. Application of 50% (v/v) Rhizopus spp. filtrate culture to the broiler chicken concentrate feed significantly reduced 82% population of A. flavus FNCC6109 after 15 days incubated relative to that of negative control (concentrate feed without addition Rhizopus spp. filtrate culture and A. flavus FNCC6109).

Immunoreactivity against Dermatophagoides pteronyssinus Assessed by the Leukocyte Adherence Inhibition Test in Patients with Intrinsic Atopic Dermatitis and Correlated “Intrinsic” Non–IgE-mediated Allergic Conditions

Background: Due to the lack of standardized laboratory procedures able to demonstrate specific immune responses against the culprit allergens, the non—IgE-mediated allergy syndromes are a group of conditions diagnosed mostly by clinical examination and exclusion criteria. Objective: To evaluate the opportunity of the Leukocyte Adherence Inhibition Test (LAIT) to discriminate specific immunoreactivity against Dermatophagoides pteronyssinus (Dp) in a group of patients with non–IgE-mediated chronic allergic conditions. Methods: Ex vivo challenge tests performed with Dp were monitored by LAIT in patients presenting diverse non–IgE-mediated allergic conditions: intrinsic Atopic Dermatitis (iAD), intrinsic Allergic Rhinitis (iAR), intrinsic Ocular Allergy (iOA), intrinsic Chronic Pharyngitis (iCP), and intrinsic Asthma (iAS). Results: The mean LAI of the control group was 7%; the mean LAI of the iAR group was 34%; the mean LAI of the iCP group was 44%; the mean LAI of the iAS group was 45%; the mean LAI of the iOA group was 47%; the mean LAI of the iAD group was 55%. The non-parametric Wilcoxon-Mann-Whitney U test comparing the control group with each other group showed significance with p-value < α = 0.05 for all groups. Conclusion: The Leukocyte Adherence Inhibition Test is an easy, quick, and inexpensive ex vivo immunoassay with the potential to predict individual immunoreactivity against HDM allergens in real-world patients with non–IgE-mediated allergies.

Isolation and antibiotic sensitivity of Campylobacter species from fecal samples of broiler chickens in North West Province, South Africa

Background and Aim: Infections with Campylobacter species have gained recognition as the most frequent cause of foodborne gastroenteritis globally. Their significance in South Africa is still an area of study interest. This study was, therefore, carried out to determine the occurrence of Campylobacter species in chickens from North West Province of South Africa as well as their antibiotic sensitivity status. Materials and Methods: A total of 2400 chicken fecal samples were collected and pooled to a total of 480 samples from five registered active poultry abattoirs in the Ngaka Modiri Molema District of North West Province, South Africa. Polymerase chain reaction (PCR) was used for the detection of Campylobacter spp. targeting the 16S rRNA gene while antibiotic sensitivity was determined using disk diffusion inhibition test. Results: After isolation, a total of 26 samples were confirmed to be harboring Campylobacter jejuni by PCR and sequencing. C. jejuni was found to be the only isolate detected in all the fecal samples tested. The study further demonstrated that C. jejuni infections were highest in the summer season (3%) followed by autumn and winter at 1%, while there were none detected in the spring. The isolated C. jejuni-positive samples on disk diffusion inhibition test displayed resistance to nalidixic acid, tetracycline, erythromycin, and ciprofloxacin at 98%, 80%, 83%, and 21%, respectively. Conclusion: C. jejuni isolated in this study is known to cause disease in humans, and thus its occurrence requires application of "One Health" strategy to reduce the spread of this zoonotic pathogen in South Africa.

Long-Term Persistence and Relevant Therapeutic Impact of High-Titer Viral-Neutralizing Antibody in a Convalescent COVID-19 Plasma Super-Donor: A Case Report

A convalescent, non-severe, patient with COVID-19 was enrolled as a hyper-immune plasma voluntary donor by the Immuno-Hematology and Transfusion Unit of the Regina Elena National Cancer Institute in Rome, under the TSUNAMI national study criteria. During a nearly 6-month period (May–October 2020), the patient was closely monitored and underwent four hyperimmune plasma collections. Serum SARS-CoV-2 (anti-S + anti-N) IgG and IgM, anti-S1 IgA, and neutralizing titers (NTs) were measured. Anti-SARS-CoV-2 antibody levels steadily decreased. No correlation was found between anti-S/anti-N IgG and IgM levels and viral NT, measured by either a microneutralization test or the surrogate RBD/ACE2-binding inhibition test. Conversely, NTs directly correlated with anti-S1 IgA levels. Hyperimmune donor plasma, administered to five SARS-CoV-2 patients with persistent, severe COVID-19 symptoms, induced short-term clinical and pathological improvement. Reported data suggest that high NTs can persist longer than expected, thus widening hyperimmune plasma source, availability, and potential use. In vitro RBD/ACE2-binding inhibition test is confirmed as a convenient surrogate index for neutralizing activity and patients’ follow-up, suitable for clinical settings where biosafety level 3 facilities are not available. IgA levels may correlate with serum neutralizing activity and represent a further independent index for patient evaluation.

Antibacterial activity of endosymbiotic bacterial compound from Pheretima sp. earthworms inhibit the growth of Salmonella Typhi and Staphylococcus aureus: in vitro and in silico approach

Background and Objectives: Earthworms coexist with various pathogenic microorganisms; thus, their immunity mecha- nisms have developed through a long process of adaptation, including through endogenous bacterial symbionts. This study aims to identify earthworm endosymbiont bacteria compounds and their antibacterial activity through an in vitro approach supported by an in silico approach. Materials and Methods: This research was conducted using the in vitro inhibition test through agar diffusion and the in silico test using molecular docking applications, namely, PyRx and Way2Drugs Prediction of Activity Spectra for Substances (PASS). Results: The in vitro results showed a potent inhibition activity with a clear zone diameter of 21.75 and 15.5 mm for Staph- ylococcus aureus and Salmonella Typhi, respectively. These results are supported by chromatography and in silico tests, which showed that several compounds in endosymbiotic bacteria, cyclo (phenylalanyl-prolyl) and sedanolide, have high binding affinity values with several antibiotic-related target proteins in both pathogenic bacteria. Cyclo (phenylalanyl-prolyl) has the highest binding affinity of -6.0 to dihydropteroate synthase, -8.2 to topoisomerase, and -8.2 to the outer membrane, whereas sedanolide has the highest binding affinity to DNA gyrase with approximately -7.3. This antibiotic activity was also clarified through the Way2Drugs PASS application. Conclusion: Ten active compounds of endosymbiont bacteria, Cyclo (phenylalanyl-prolyl) and sedanolide were potential candidates for antibacterial compounds based on the inhibition test of the agar diffusion method and the results of reverse docking and Way2Drugs PASS.

A Preliminary Study on Cross-Reactivity of Heat-Treated Quail and Hen’s Egg White Proteins in Young Children

We investigated the effects of different types of heat treatments on hen’s egg white (HEw) and quail egg white (QEw) proteins and their cross-reactivity in young children. Crude extracts of raw and water-boiled HEw and QEw and commercially developed stone-baked HEw were prepared. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) was then performed. Patients diagnosed with HEw allergy were enrolled, and pooled sera were tested with each extract in an enzyme-linked immunosorbent assay (ELISA)-inhibition test. A skin prick test (SPT) and oral food challenge (OFC) were also performed. The median age of 12 patients was 2.5 years. SDS-PAGE results revealed strongly stained bands for the ovomucoid of boiled HEw and QEw, while stone-baked HEw displayed remarkable changes for all protein fractions. In the ELISA-inhibition test, pre-incubation of the sera led to a profound decrease, moderate decrease, and minimal decrease in the amount of IgE binding to boiled and raw HEw, boiled and raw QEw, and stone-baked HEw proteins, respectively. SPTs and OFC demonstrated cross-reactivity values of 41.7% (5/12) and 16.7% (2/12) for boiled QEw and stone-baked HEw, respectively. We observed moderate cross-reactivity between QEw and HEw. Boiling had a limited effect on altering egg allergenicity. Commercially developed, stone-baked HEw can be an alternative food for children with HE allergy.

Assessment of Immunoreactivity against Therapeutic Options Employing the Leukocyte Adherence Inhibition Test as a Tool for Precision Medicine

Background: The Precision Medicine’s approach employs the endotype concept as a central feature to personalize medical treatment. Individual immunoreactivity, alongside characteristics such as genetics, environment, and diet, is one of the factors that differentiates the therapeutic-driven endotypes. Objective: To evaluate the opportunity of the Leukocyte Adherence Inhibition test to differentiate the immunoreactivity between two similar therapeutic agents employed on Allergen Immunotherapy. Methods: Side by side Leukocyte Adherence Inhibitions tests were performed with ovalbumin and carbamylated ovalbumin on a population of 33 self-reported egg-allergic individuals. Results: The results showed two endotypes inside the immune response of the studied groups: The first endotype was defined by the 16 individuals that presented a significant decrease in ovalbumin’s immunoreactivity after carbamylation (mean of differences = 35%; p = 0.002). The second endotype was defined by 17 individuals that presented a significant increase in ovalbumin’s immunoreactivity after carbamylation (mean of differences = 32%; p = 0.001). Conclusion: The Leukocyte Adherence Inhibition test was able to differentiate two distinct immunoreactivity patterns when comparing two similar therapeutic agents suggesting, as proof of concept, a potential role to be employed as a Precision Medicine tool.