Emergence and Genomic Characterization of a spa Type t4407 ST6-SCCmec Type IVa Methicillin-Resistant Staphylococcus aureus Strain Isolated from Al-Karak Hospital, Jordan.

Background and Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) is a major concern in Jordanian hospitals in terms of infection control. The purpose of this study was to identify the resistance patterns of Staphylococcus aureus strains isolated from surfaces of critical locations within the Al-Karak Governmental Hospital in 2019. Additionally, the study aimed to conduct whole-genome sequencing on the isolates. Materials and Methods: In February 2019, fourteen S. aureus strains were isolated from surfaces in critical sites in the Al-Karak Governmental Hospital. These isolates underwent antibiogram testing to determine their resistance profile. Genome sequencing using the Illumina MiSeq platform was applied to the extracted DNA from these isolates. The genomic data, including coding sequences, were analyzed to identify lineage, resistance genes, and plasmids. Results: The antibiogram results revealed that 11 of the 14 isolates were resistant to oxacillin, 6 to linezolid, and 1 to rifampicin, while none showed resistance to chloramphenicol. Eleven isolates were identified as MRSA, with a novel spa type (t4407) not previously reported in Jordan. High-quality sequencing data were obtained for only one isolate, i.e., A29, the genome showed 2,789,641 bp with a 32.7% GC content and contained 2650 coding sequences. Genomic analysis indicated the ST6 lineage, mecA gene (SCCmec type IVa(2B)), and a hybrid plasmid (pJOR_blaZ) carrying the blaZ gene for ß-lactam resistance. Genomic data were deposited in NCBI (CP104989). The A29 genome closely resembled an MRSA genome isolated from a Danish hospital in 2011. The SNP analysis revealed identical antimicrobial resistance genes in these two genomes. Conclusions: This study unveils the first genomic sequence of an MRSA isolate from Jordan, marked by distinctive genotypic traits. The findings enhance our understanding of the MRSA types circulating in Jordan and the region and substantiate the phenomenon of intercontinental MRSA transmission.

A combined inactivated cholera and hepatitis A vaccine-induced potent protective immunity in a mouse model.

Cholera and hepatitis A are serious infections spread by consuming contaminated food or water. Vaccination is the most effective strategy to prevent them. Inactivated vaccines are available for both diseases. Our goal in this study is to evaluate the immunogenic response of hepatitis A and cholera combination vaccines compared to the separate vaccines. Hepatitis A and cholera vaccine formulations with and without adjuvants (alum or chitosan) were developed and injected into mice intraperitoneally. We measured the rate of seroconversion; serum-specific antibody titers; lymphoproliferation analysis; cytokine secretions for IL2, IL4, IL10, and IFN-; and a challenge test against cholera strains in the vaccinated mice. Based on the results, the combined vaccination formulation, whether adjuvanted or not, significantly boosted the immune response on both humoral and cellular levels against both hepatitis A and cholera antigens compared to the individual vaccines. These findings validated an important concept for developing an effective combined cholera and hepatitis A vaccine that could be introduced as a novel combined vaccine for travelers as part of a standard immunization schedule. KEY POINTS: • Cholera and hepatitis A combined vaccines (with or without adjuvants) were prepared. • The vaccines were injected into mice groups for humoral and cellular immunity evaluation. • Combined vaccines gave substantial protection against both immunogens.

Evaluation of MMR live attenuated vaccine oncolytic potential using Ehrlich ascites carcinoma in a murine model.

MMR vaccine is a common vaccine that contains oncolytic viruses (Measles, Mumps, and Rubella) and could be used as a potential anti-cancer treatment. In this study, we assessed the anti-tumor activity of the MMR vaccine against Ehrlich ascites carcinoma (EAC) solid tumor induced in mice. The in vitro assay showed that vaccine IC50 in EAC was approximately 200 CCID50. The vaccine was intratumorally administrated twice weekly in EAC-bearing mice. The antitumor response of the vaccine was measured by tumor growth, survival rate, histopathologic examination, flow cytometry analysis, and body biochemical parameters. The MMR vaccine demonstrated a substantial reduction of tumor growth and prolongation of life span as well. The proliferation marker was significantly lower in the vaccine-treated group. Moreover, the apoptosis key parameter Casp-3 was also higher in the vaccine-treated group. The vaccine somewhat restored the deterioration of the biochemical parameters (LDH, GOT, GPT, MDA, NO, and PON-1) in the tumor-bearing mice. Finally, this study indicated the potential antitumor effect of MMR vaccine via anti­proliferative, apoptotic activities, and modulating the antioxidant parameters. This study opens a new field of inquiry for future research on the vaccine's anti-cancer properties.

Comparative evaluation of the humoral immune interaction when BCG and conjugated meningococcal vaccines combined or co-administrated in mice.

BCG and conjugated meningococcal vaccines (cMen) are part of the recommended immunization program for young children (<5 years) in many countries all over the world. However, there was no immunogenicity supportive data to assess the effect of BCG and cMen vaccines when combined or co-administrated. Therefore, we sought to evaluate the antibody response in mice groups when BCG and cMen vaccines were whether in combination, co-administration simultaneously or administration with a 14-day gap interval. Our results showed that cMen either combined with BCG vaccine or co-administrated had a significant negative humoral immune effect on each other. This negative impact was observed also when there were 2 weeks between their administration regardless of the vaccination order. Therefore, our results support that it is preferable to separate the two vaccination for > 14 days at least. But additional studies are needed to explore the cellular-mediate response of the immune intervention as well.

Chitosan and alginate salt as biomaterials are potential natural adjuvants for killed cholera vaccine.

Chitosan and alginate salts are natural biopolymers that have gained recent attention in the biomedical sectors. Their properties allow them to become potential candidates as safe, cheap, and effective vaccine adjuvants. The present study aimed to enhance the immunogenic response of a current injectable killed cholera vaccine (KCV) using chitosan and alginate salt as natural adjuvants against alum. We tested KCV adjuvanted with alum, chitosan, and sodium alginate in mice. Mice were immunized intraperitoneally with KCV adjuvanted with alum, chitosan, or alginate salt and compared with a control unadjuvanted immunized group. Humoral, cellular, and functional immune responses were evaluated in all groups. The addition of adjuvants, particularly natural adjuvants, to KCV significantly improved the immune response as demonstrated by specific antibody increase, strong proliferation effects, and high protection rate against different challenge doses of cholera strains. Our findings demonstrate that chitosan and alginate salt are superior adjuvants for boosting the KCV immune response and highlights the requirement for further vaccine development.

Alginate-coated chitosan nanoparticles act as effective adjuvant for hepatitis A vaccine in mice.

The numerous recent hepatitis A outbreaks emphasize the need for vaccination; despite the effectiveness of the current ones, developments are needed to overcome its high cost plus some immune response limitations. Our study aims to evaluate the use of chitosan and alginate-coated chitosan nanoparticles as an adjuvant/carrier for the hepatitis A vaccine (HAV) against the traditional adjuvant alum. Immune responses towards (HAV-Al) with alum, (HAV-Ch) with chitosan, and (HAV-aCNP) with alginate-coated chitosan nanoparticles, were assessed in mice. HAV-aCNP significantly improved the immunogenicity by increasing the seroconversion rate (100%), the hepatitis A antibodies level, and the splenocytes proliferation. Thus, the HAV-aCNP adjuvant was superior to other classes in IFN-γ and IL-10 development. Meanwhile, the solution formula of HAV with chitosan showed comparable humoral and cellular immune responses to alum-adjuvanted suspension with a balanced Th1/Th2 immune pathway. The current study showed the potential of alginate-coated chitosan nanoparticles as an effective carrier for HAV. Consequently, this would impact the cost of HAV production positively.

Chitosan and Sodium Alginate Combinations Are Alternative, Efficient, and Safe Natural Adjuvant Systems for Hepatitis B Vaccine in Mouse Model.

Hepatitis B viral (HBV) infections represent major public health problem and are an occupational hazard for healthcare workers. Current alum-adjuvanted HBV vaccine is the most effective measure to prevent HBV infection. However, the vaccine has some limitations including poor response in some vaccinee and being a frost-sensitive suspension. The goal of our study was to use an alternative natural adjuvant system strongly immunogenic allowing for a reduction in dose and cost. We tested HBV surface antigen (HBsAg) adjuvanted with chitosan (Ch) and sodium alginate (S), both natural adjuvants, either alone or combined with alum in mouse model. Mice groups were immunized subcutaneously with HBsAg adjuvanted with Ch or S, or triple adjuvant formula with alum (Al), Ch, and S, or double formulations with AlCh or AlS. These were compared to control groups immunized with current vaccine formula or unadjuvanted HBsAg. We evaluated the rate of seroconversion, serum HBsAg antibody, IL-4, and IFN-γ levels. The results showed that the solution formula with Ch or S exhibited comparable immunogenic responses to Al-adjuvanted suspension. The AlChS gave significantly higher immunogenic response compared to controls. Collectively, our results indicated that Ch and S are effective HBV adjuvants offering natural alternatives, potentially reducing dose.