Antibiotic resistance of Mycoplasma Synoviae strains isolated in China from 2016 to 2019

Background In the past decade, Mycoplasma synoviae (M. synoviae) infection has become widely prevalent in China, has caused serious economic losses and has become one of the most important diseases in the chicken industry. Medication is a general approach for the control of M. synoviae infection, but antibiotics are sometimes ineffective in clinical practice. To investigate the sensitivity of M. synoviae to antimicrobials commonly used in the treatment of M. synoviae infection, the antibiotic susceptibility of 32 M. synoviae strains isolated from China from 2016 to 2019 were determined using the minimum inhibitory concentration (MIC) method. Results All isolates had low MIC values for the combination of lincomycin and spectinomycin, pleuromutilin, and macrolides. However, the M. synoviae isolates displayed variance in MICs for doxycycline hydrochloride with a range of 0.25 to 8 μg/mL, and oxytetracycline hydrochloride with a range of 0.5 to 8 μg/mL. Three and one M. synoviae isolates showed intermediate MIC values to doxycycline hydrochloride and oxytetracycline hydrochloride, respectively. High MIC values for enrofloxacin were detected in all isolates with MICs ranging from 4 to 32 μg/mL. Furthermore, comparison of the parC QRDR identified a mutation at nucleotide position 254 (C254T) resulting in a Thr 85 Ile amino acid change in all M. synoviae isolates and the reference strain ATCC 25204 being resistant to enrofloxacin. Moreover, mutations at Glu 804 Gly and Thr 686 Ala of gyrA QRDR were identified in all M. synoviae isolates and ATCC 25204. The mutation in the QRDR of the parE gene resulted in amino acid changes at positions 197 (Pro to Ser) in 27/32 M. synoviae isolates. Conclusion Three nonsynonymous mutations in gyrA and parE were first identified to be related to enrofloxacin resistance. Our results showed that M. synoviae resistance to enrofloxacin is widespread.

Neutralization of SARS-CoV-2 Omicron pseudovirus by BNT162b2 vaccine-elicited human sera

A new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineage, B.1.1.529, was recently detected in Botswana and South Africa and is now circulating globally. Just two days after it was first reported to the World Health Organization (WHO), this strain was classified as a variant of concern (VOC) and named Omicron. Omicron has an unusually large number of mutations, including up to 39 amino acid modifications in the spike (S) protein, raising concerns that its recognition by neutralizing antibodies from convalescent and vaccinated individuals may be severely compromised. In this study, we tested pseudoviruses carrying the SARS-CoV-2 spike glycoproteins of either the Wuhan reference strain, the Beta, the Delta or the Omicron variants of concern with sera of 51 participants that received two doses or a third dose (≥6 months after dose 2) of the mRNA-based COVID-19 vaccine BNT162b2. Immune sera from individuals who received two doses of BNT162b2 had more than 22-fold reduced neutralizing titers against the Omicron as compared to the Wuhan pseudovirus. One month after a third dose of BNT162b2, the neutralization titer against Omicron was increased 23-fold compared to two doses and antibody titers were similar to those observed against the Wuhan pseudovirus after two doses of BNT162b2. These data suggest that a third dose of BNT162b2 may protect against Omicron-mediated COVID-19, but further analyses of longer-term antibody persistence and real-world effectiveness data are needed.

BIOSURFACTANTS SYNTHESIS BY PSEUDOMONAS AERUGINOSA BACTERIA ISOLATED FROM THE SURFACE OF MUSSELS OF THE BLACK SEA

Aim. Establishing of the ability to synthesize surface-active compounds by Pseudomonas aeruginosa bacteria isolated from the surface of Black Sea mussels. Methods. During the research several marine Pseudomonas spp strains isolated from petroleum hydrocarbon contaminated areas of Black Sea wereused: P. aeruginosa M1, P. aeruginosa M4 and P. aeruginosa PA01 as reference strain in suspension and biofilm cultures (LB and Giss media). Cultivation of Pseudomonas aeruginosa strains was performed at 37 °C for 120 and 168 hours. Planktonic culture growth was determined spectrophotometrically on the wave length 600 nm. Biofilm mass was determined spectrophotometrically on the wave length 592 nm by CV-test. The presence of surface-active compounds was determined in a drop test. The quantitative content of rhamnolipids was evaluated by the color reaction of rhamnose with orcin. Results. P. aeruginosa strains M1 and M4 isolated from Black Sea mussel’s surfaces synthesize 25% and 66% more surfactants than the reference strain PA01. All strains in Giss medium synthesized 10–20 times less rhamnolipids than in LB medium. In biofilm cultures the same biosurfactant synthesis dependence on the composition of the nutrient medium is observed as in suspension cultures. According to the intensity of rhamnolipid production in biofilm cultures, the studied strains can be arranged in the following row: P. aeruginosa M4 > P. aeruginosa M1 >> P. aeruginosa PA01.Conclusions. The strains of P. aeruginosa isolated from the Black Sea are more efficient producers of rhamnolipids than the reference strain of P. aeruginosa PA01; the intensity of biosynthesis of surfactants significantly depends on the composition of the nutrient medium and the method of cultivation.

In vitro effect of salinity and pH on Fusarium sp., the causal agent of sweet-potato root rot

Fusarium root rot in a common pathogen of sweet potato, with a wide range of host plants. In the current study six new isolates of Fusarium sp., collected from infected sweet potato plants, along with a reference strain of Fusarium oxysporum, had their growth behavior studied in various pH and saline conditions. In vitro studies showed that salinity higher than 6% NaCl in the PDA substrate significantly reduces the fungal growth. At 12% NaCl, four of seven strains revealed complete mycelia inhibition. However, for the other two isolates, and for the reference strain, 12% salinity only reduced the growth with 77.4%. Regarding the fungal growth at different pH values, it was noticed that tested fusaria were not perturbed at up to 8.5 alkalinity. However, at a pH of 4.5, the growth rate was reduced, although the growth differences were diminished during prolonged incubation time. Considering the in vitro results, saline water should be tested as preventive immersion treatment on the sweet potato sprouts, before their planting, in order to reduce the incidence of Fusarium infection.

Modeling the onset of symptoms of COVID-19: Effects of SARS-CoV-2 variant

Identifying order of symptom onset of infectious diseases might aid in differentiating symptomatic infections earlier in a population thereby enabling non-pharmaceutical interventions and reducing disease spread. Previously, we developed a mathematical model predicting the order of symptoms based on data from the initial outbreak of SARS-CoV-2 in China using symptom occurrence at diagnosis and found that the order of COVID-19 symptoms differed from that of other infectious diseases including influenza. Whether this order of COVID-19 symptoms holds in the USA under changing conditions is unclear. Here, we use modeling to predict the order of symptoms using data from both the initial outbreaks in China and in the USA. Whereas patients in China were more likely to have fever before cough and then nausea/vomiting before diarrhea, patients in the USA were more likely to have cough before fever and then diarrhea before nausea/vomiting. Given that the D614G SARS-CoV-2 variant that rapidly spread from Europe to predominate in the USA during the first wave of the outbreak was not present in the initial China outbreak, we hypothesized that this mutation might affect symptom order. Supporting this notion, we found that as SARS-CoV-2 in Japan shifted from the original Wuhan reference strain to the D614G variant, symptom order shifted to the USA pattern. Google Trends analyses supported these findings, while weather, age, and comorbidities did not affect our model’s predictions of symptom order. These findings indicate that symptom order can change with mutation in viral disease and raise the possibility that D614G variant is more transmissible because infected people are more likely to cough in public before being incapacitated with fever.

A Case Report: Genetically Distinct Severe Acute Respiratory Syndrome Coronavirus-2 Variant Causing Reinfection

Background: The emergence of novel variants has been a great deal of international concern since the recently published data suggest that previous infections with SARS-CoV-2 may not protect an individual from new variants. We report a patient had two distinct episodes of COVID-19 with different variants of SARS-CoV-2.Methods: The nasopharyngeal samples collected from the two episodes were subjected to whole-genome sequencing and comparative genome analysis.Results: The first infection presented with mild symptoms, while the second infection presented with severe outcomes which occurred 74 days after the patient recovered from the first episode. He had elevated C-reactive protein, ferritin, and bilateral consolidation as a sign of acute infection. Genome analysis revealed that the strains from the first and second episodes belonged to two distinct Nexstrain clades 20B and 20I and Pangolin lineages B.1.1.25 and B.1.1.7, respectively. A total of 36 mutations were observed in the episode-2 strain when compared with the reference strain Wuhan-Hu-1. Among them, eight mutations were identified in the receptor-binding domain (RBD).Conclusion: Our findings concern whether the immunity acquired by natural infection or mass vaccination could confer adequate protection against the constantly evolving SARS-CoV-2. Therefore, continuous monitoring of genetic variations of SARS-CoV-2 strains is crucial for interventions such as vaccine and drug designs, treatment using monoclonal antibodies, and patient management.

Viability, Stability and Biocontrol Activity in Planta of Specific Ralstonia Solanacearum Bacteriophages After Their Conservation Prior to Commercialization and Use

Ralstonia solanacearum is a pathogen that causes bacterial wilt producing severe damage in staple solanaceous crops. Traditional control has low efficacy and/or environmental impact. Recently, the bases of a new biotechnological method by lytic bacteriophages vRsoP-WF2, vRsoP-WM2 and vRsoP-WR2 with specific activity against R. solanacearum were established. However, some aspects remain unknown, such as the survival and maintenance of the lytic activity after submission to a preservation method as the lyophilization. To this end, viability and stability of lyophilized vRsoP-WF2, vRsoP-WM2 and vRsoP-WR2 and their capacity for bacterial wilt biocontrol have been determined against one pathogenic Spanish reference strain of R. solanacearum in susceptible tomato plants in different conditions and making use of various cryoprotectants. The assays carried out have shown satisfactory results with respect to the viability and stability of the bacteriophages after the lyophilization process, maintaining high titres throughout the experimental period, also with respect to the capacity of the bacteriophages for the biological control of bacterial wilt, controlling this disease in more than 50% of the plants. The results offer good prospects for the use of lyophilization as a conservation method for the lytic bacteriophages of R. solanacearum in view of their commercialization as biocontrol agents.

DETERMINACIÓN DE LA CONCENTRACIÓN EFECTIVA 50, 90 Y 99 DE IVERMECTINA SOBRE UNA CEPA DE GARRAPATA COMÚN DEL BOVINO (Rhipicephalus (Boophilus) microplus) PROVENIENTE DE LA PROVINCIA DE SANTA FE-ARGENTINA

The aim of this study was to obtain the sensitivity data of the native strains of R. microplus to ivermectin (IVM) in the province of Santa Fe-Argentina, in order to detect the emergence of resistant strains. Native (field isolation) and susceptible (reference) larvae of R. microplus were treated with increasing concentrations of IVM in order to determine its efficacy at different doses. The statistical treatment of the concentration-efficacy curves was performed by the PROBIT model, who is a standardized method proposed by FAO. The results obtained indicated that the larvae isolated were susceptible, being the values of LD50 (9.66 ppm) and LD90 (17.41 ppm), which were lower than those of the reference strain (LD50:13.58 ppm, LD90:19.31 ppm), and the RR (resistance ratio) were less than one (RR50: 0.711; RR90: 0.901). The DL99 (51.48 ppm) was only obtained in the reference strain to determine the discriminant dose (DL99x2), which is a parameter that could be used to easily discriminate susceptible and resistant larvae populations.

EFECTO INHIBITORIO DE Lactobacillus plantarum AISLADOS DESDE LAS DISTINTAS ETAPAS DE LA CADENA PRODUCTIVA DE CARNE PORCINA FRENTE A Campylobacter coli

Lactobacillus plantarum has a great capacity to adapt to environmental and have antibacterial capacity against different pathogen. This is a potential alternative for the control of pathogen in food coming slaughter animal. The objective of this work was to determine the antibacterial effect of L. plantarum strains against C. coli strains. L. plantarum LP1, LP3, LP5 and LP7 of porcine origin and a reference strain of Campylobacter coli NCTC 11366 were used. C. coli and L. plantarum was reactivated. Lyophilized ELC and ELCn were obtained. C. coli was distributed on the surface of mCCDA by swab and the ELC and the ELCn were inoculated in wells previously made on the agar. After 48 h, the diameter of the zone of inhibition was measured. The inhibition halos produced were 23.2 mm. The ELCn showed less antagonistic activity. The LP5 ELC generated higher inhibition halos and ELCn showed no differences between the four strains studied. L. plantarum strains, isolated from the pig production chain, could potentially be applied to control C. coli. This data is added to the scarce scientific evidence of the inhibitory effect of L. plantarum before this zoonotic pathogen of importance in public health.

MOLECULAR-GENETIC CHARACTERIZATION OF PARAINFLUENZA-3 VIRUS ISOLATES

This study reports on the molecular-genetic characterization of a field strain of BPIV-3 isolated in the Republic of Tatarstan. M gene fragments of the Russian isolate LD-9 and the vaccine strain PTK/86 were sequenced and aligned with the corresponding sequences of the virus strains presented in GenBank. The results of the phylogenetic analysis showed that the Russian isolate LD-9 (MW52481) and the vaccine strain PTK45/86 belong to the genotype A of the parainfluenza virus-3, as well as the reference strain SF-4. The Russian isolate LD-9 showed 98.3 % similarity with the reference strain SF-4, and the Russian vaccine strain PTK-45/86 was 100 % identical to the reference strain.