Gut and Colony Microbiota of Honey Bees: Social Immunity and Opportunism Overwinter

Overwintering is a major contributor to honey bee colony loss and involves factors that influence disease susceptibility. Honey bees possess a secretory head gland that interfaces with the extended social environment on many levels. With the coming of winter, colonies produce a long-lived (diutinus) worker phenotype that survives until environmental conditions improve. We used a known-age worker cohort to investigate microbiome integrity and social gene expression of diutinus workers overwinter. We provide additional context by contrasting host-microbial interactions from warm outdoor and cold indoor overwintering environments. Our results provide the first evidence that social immune gene expression is associated with diutinus bees, and highlight the midgut as a target of opportunistic disease overwinter. Host microbial interactions suggest opportunistic disease progression and resistance in diutinus workers, but susceptibility to opportunistic disease in younger workers that emerged during the winter, including increases in Enterobacteriaceae, fungal load and bacterial diversity abundance. The results are consistent with increased social immunity overwinter, including host associations with the colony microbiota, and a social immune response by long-lived diutinus workers to combat microbial opportunism. The cost/benefit ratio associated with limited expression of the diutinus phenotype may be a strong determinant of colony survival overwinter.

Clinical Spectrum and Resistance Profile of Staphylococcus Infections in a Peri Urban Tertiary Care Hospital

Staphylococcus may be evaluated to be a serious microorganism to colonizes and contaminate both the healthy and immuno-competent people in the community along with the hospitalized patients with decreased immunity. This bacterium has been commonly present on the skin and in the nasal cavity of the human body. In the particular sites the organisms may give rise to local diseases of the nose, urethra, vagina and gastrointestinal tract, skin however most of them are minor and not associated with any mortality. S. aureus as well as coagulase-negative staphylococcus which are gram-positive bacteria that lives on the upper respiratory, mouth and skin structure creation, are the possible element for nosocomial and opportunistic disease in humans as well as in animals. Major complications involved with staphylococcus involved SSI, bullous impetigo, skin & soft tissues infection, bacteremia, pulmonary infections etc. The situation is further complicated by methicillin resistance in staphylococcus. MRSA/MRCoNS are considered as the emerging cause of hospital-acquired infections. it is very necessary to quantify the load of infections in order to achieve good hospital infection control policies. Furthermore, the morbidity and mortality rate may also be reduced in the same manner.

X-linked immunodeficient (XID) mice exhibit high susceptibility to Cryptococcus gattii infection

Cryptococcosis is an opportunistic disease caused by the fungus Cryptococcus neoformans and Cryptococcus gattii. It starts as a pulmonary infection that can spread to other organs, such as the brain, leading to the most serious occurrence of the disease, meningoencephalitis. The humoral response has already been described in limiting the progression of cryptococcosis where the B-1 cell seems to be responsible for producing natural IgM antibodies, crucial for combating fungal infections. The role of the B-1 cell in C. neoformans infection has been initially described, however the role of the humoral response of B-1 cells has not yet been evaluated during C. gattii infections. In the present study we tried to unravel this issue using XID mice, a murine model deficient in the Btk protein which compromises the development of B-1 lymphocytes. We use the XID mice compared to BALB/c mice that are sufficient for the B-1 population during C. gattii infection. Our model of chronic lung infection revealed that XID mice, unlike the sufficient group of B-1, had early mortality with significant weight loss, in addition to reduced levels of IgM and IgG specific to GXM isolated from the capsule of C. neoformans. In addition to this, we observed an increased fungal load in the blood and in the brain. We described an increase in the capsular size of C. gattii and the predominant presence of cytokines with a Th2 profile was also observed in these animals. Thus, the present study strongly points to a higher susceptibility of the XID mouse to C. gattii, which suggests that the presence of B-1 cells and anti-GXM antibodies is fundamental during the control of infection by C. gattii.

Gut and Colony Microbiota of Honey Bees Apis mellifera: Social Immunity, Opportunistic Disease and Survival Overwinter.

BackgroundOverwintering is a major contributor to honey bee colony loss and involves changes in environmental conditions, host physiology and group behavior that influence disease susceptibility. Honey bees possess a secretory head gland that interfaces with the extended colony environment on many levels, producing pro-oxidants, antioxidants and antimicrobial peptides. With the coming of winter, colonies produce a long-lived (diutinus) worker phenotype that survives until environmental conditions improve. We used a known-age worker cohort to investigate microbiome integrity and social gene expression of diutinus workers overwinter. We provide additional context by contrasting host-microbial interactions from warm outdoor and cold indoor overwintering environments. ResultsWe produce the first evidence that social immune gene expression is associated with the core hindgut and colony microbiota in honey bees, and highlight the midgut as a target of opportunistic disease overwinter. We discovered a distinct physiological and microbiological trajectory for diutinus workers that differs drastically from younger, short-lived workers in the colony. Diutinus bees were associated with decreased fungal load and decreased bacterial diversity, and increased core microbiota and longevity. Colonies overwintered indoors maintained a stable or improved microbiota structure and complimentary gene expression overwinter. In contrast, workers from colonies overwintered outdoors in warm southern conditions possessed changes co-occurring throughout the alimentary tract microbiota that suggest opportunistic disease progression and resistance in diutinus workers, but susceptibility to opportunistic disease in younger workers that emerged during the winter, including increases in Enterobacteriaceae, fungal load and bacterial diversity abundance. ConclusionsOur results highlight social selection pressures that shaped the colony and hindgut microbiome with evolution to a perennial life history. The results are consistent with a “group level” explanation of social immunity, including host associations with the colony microbiota, and a social immune response by long-lived diutinus workers to accompany microbial opportunism. The cost/benefit ratio associated with limited expression of the diutinus phenotype may be a strong determinant of colony survival overwinter. The relationship of colony and gut microbiota with social immune function highlights the range of host-microbial interaction associated with the honey bee superorganism, and its potential influence on colony health, disease resistance and gut integrity.

Microflora of autopsy material from HIV-infected patients

According to literature, lungs are affected in 25 – 60% of HIV-infected individuals. The most common opportunistic disease in this group of patients is pneumonia of various etiology. HIV-positive patients show a wide microbiological landscape of pneumonia pathogens.The aim of the study was to determine the structure of the microflora of autopsy material obtained from the lungs of HIV-infected patients.Methods. 83 samples of autopsy material from patients with HIV infection and 27 samples from HIV-negative patients were analyzed. The deceased were treated in the pulmonary departments of Samara with a diagnosis of community-acquired pneumonia. Due to the large variety of microorganisms recovered from the autopsy material, the microorganisms were divided into several groups: Enterobacteria, non-fermentative gram-negative bacteria, Enterococci, Staphylococci, Sterptococci, and Corynebacteria. The fungi were divided into two subgroups. The first included Candida spp., the second included other types of fungi. The tests for pneumocystis infection, Mycobacterium tuberculosis, and non-tuberculosis mycobacteria were not performed.Results. Enterobacteria were found in 39.2%, non-fermentative gram-negative bacteria – in 27.3% and Enterococci – in 19.6% of the HIV-infected patients. Staphylococcus spp. were found in 4.2%, and Corynebacteria and Streptococci in 0.7% of those patients. Candida spp. accounted for 6.3%, other fungi were found in 2.0% of cases. Enterobacteria and non-fermentative gram-negative bacteria were found in 26.2% of HIV-negative patients. Enterococcus spp. – 21.4%, Staphylococci – 7.1%, Streptococci – 2.4% of those patients. Candida spp. 16.7% were found in 16.7% of HIV-negative patients. Enterobacteria were found significantly more often in the autopsy material from HIV-infected patients. The study shows the high importance of gram-negative flora as a causative agent of infectious lesions of the lung tissue. At the same time, we statistically confirmed an increase in prevalence of Enterobacteria in HIV-positive patients. Although these pathogens are not always detected in the sputum from HIV-infected patients with bronchopulmonary disorders, they are detected only by autopsy examination of the lung tissue.Conclusion. Diagnosis and treatment of pneumonia in HIV-infected patients with severe immunodeficiency have a few specific features, such as increasing the frequency of sputum culture, reducing the number of errors at the preanalytical stage of collecting the biomaterial, ensuring proper conditions for its transportation to a bacteriological laboratory, improving the quality of analysis. The physicians should be aware of the possible detection of gram-negative flora as a causative agent of lung infections and the possible change of the causative agent.

m6A RNA methylation facilitates pre-mRNA 3’-end formation and is essential for viability of Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular parasite that can cause serious opportunistic disease in the immunocompromised or through congenital infection. To progress through its life cycle, Toxoplasma relies on multiple layers of gene regulation that includes an array of transcription and epigenetic factors. Over the last decade, the modification of mRNA has emerged as another important layer of gene regulation called epitranscriptomics. Here, we report that epitranscriptomics machinery exists in Toxoplasma, namely the methylation of adenosines (m6A) in mRNA transcripts. We identified novel components of the m6A methyltransferase complex and determined the distribution of m6A marks within the parasite transcriptome. m6A mapping revealed the modification to be preferentially located near the 3’-boundary of mRNAs. Knockdown of the m6A writer components METTL3 and WTAP resulted in diminished m6A marks and a complete arrest of parasite replication. Furthermore, we examined the two proteins in Toxoplasma that possess YTH domains, which bind m6A marks, and showed them to be integral members of the cleavage and polyadenylation machinery that catalyzes the 3’-end processing of pre-mRNAs. Loss of METTL3, WTAP, or YTH1 led to a defect in transcript 3’-end formation. Together, these findings establish that the m6A epitranscriptome is essential for parasite viability by contributing to the processing of mRNA 3’-ends.

Molecular epidemiology of Microsporidia among HIV-positive and HIV-negative patients in the Limpopo province, South Africa

Introduction: Human microsporidiosis represents an important and rapidly emerging opportunistic disease. The present study investigated the prevalence of microsporidia among HIV positive and HIV negative patients with or without diarrhoea in Vhembe and Mopani Districts in the Limpopo Province. Methodology: A total of 170 stool samples were collected from these patients and microsporidia species was detected using a Real-Time PCR targeting a conserved region of the small ribosomal subunit rRNA (SSU-rRNA) gene of Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi. Results: Fifty six (32.9%) were positive for microsporidia. The prevalence was higher in HIV negative patients (36.6%) while 24.1% of patients who were HIV positive had microsporidia. Microsporidia was more common among patients aged between 1 and 10 years (52.6%). However among the HIV positive patients, microsporidia prevalence was higher among those that were not taking antiretrovirals (ARVs) compared to those who were on ARVs, (36.6%) and (24.1%), respectively. Microsporidia was also noted to be significantly associated with diarrheal and stomach pains; p = 0.02 and p = 0.048, respectively. Furthermore, microsporidia infections was more prevalent among patients who had animals at home (p = 0.037). Conclusions: Study has shown a high prevalence of microsporidia among patients attending primary health centers in the Mopani District for the first time. Prevalence of microsporidia was higher among HIV negative and HIV positive patients who were not on ARV treatment. Keeping animals in the household appeared to be a risk of getting infected with microsporidia. Further studies are needed to determine the genetic characteristics of these organisms in the study population.

Structural insights into the intermolecular interaction of the adhesin SdrC in the pathogenicity of Staphylococcus aureus

Staphylococcus aureus is an opportunistic disease-causing pathogen that is widely found in the community and on medical equipment. A series of virulence factors secreted by S. aureus can trigger severe diseases such as sepsis, endocarditis and toxic shock, and thus have a great impact on human health. The transformation of S. aureus from a colonization state to a pathogenic state during its life cycle is intimately associated with the initiation of bacterial aggregation and biofilm accumulation. SdrC, an S. aureus surface protein, can act as an adhesin to promote cell attachment and aggregation by an unknown mechanism. Here, structural studies demonstrate that SdrC forms a unique dimer through intermolecular interaction. It is proposed that the dimerization of SdrC enhances the efficiency of bacteria–host attachment and therefore contributes to the pathogenicity of S. aureus.

m6A RNA methylation facilitates pre-mRNA 3’-end formation and is essential for viability of Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular parasite that can cause serious opportunistic disease in the immunocompromised or through congenital infection. To progress through its life cycle, Toxoplasma relies on multiple layers of gene regulation that includes an array of transcription and epigenetic factors. Over the last decade, the modification of mRNA has emerged as another important layer of gene regulation called epitranscriptomics. Here, we report that epitranscriptomics machinery exists in Toxoplasma, namely the methylation of adenosines (m6A) in mRNA transcripts. We identified novel components of the m6A methyltransferase complex and determined the distribution of m6A marks within the parasite transcriptome. m6A mapping revealed the modification to be preferentially located near the 3’-boundary of mRNAs within the consensus sequence, YGCAUGCR. Knockdown of the m6A writer enzyme METTL3 resulted in diminished m6A marks, loss of a target transcript, and a complete arrest of parasite replication. Furthermore, we examined the two proteins in Toxoplasma that possess YTH domains, which bind m6A marks, and showed them to be integral members of the cleavage and polyadenylation machinery that catalyzes the 3’-end processing of pre-mRNAs. Together, these findings establish that the m6A epitranscriptome is essential for parasite viability by contributing to the processing of mRNA 3’-ends.Author SummaryToxoplasma gondii is a parasite of medical importance that causes disease upon immuno-suppression. Uncovering essential pathways that the parasite uses for its basic biological processes may reveal opportunities for new anti-parasitic drug therapies. Here, we describe the machinery that Toxoplasma uses to modify specific adenosine residues within its messenger RNAs (mRNA) by N6-adenosine methylation (m6A). We discovered that m6A mRNA methylation is prevalent in multiple stages of the parasite life cycle and is required for parasite replication. We also establish that m6A plays a major role in the proper maturation of mRNA. Two proteins that bind m6A modifications on mRNA associate with factors responsible for the cleavage and final processing steps of mRNA maturation. Since all of the machinery is conserved from plants to Toxoplasma and other related parasites, we propose that this system operates similarly in these organisms.

A Case of Pneumocystis jirovecii Pneumonia under Belatacept and Everolimus: Benefit-Risk Balance between Renal Allograft Function and Infection

<i>Pneumocystis jirovecii</i> pneumonia is an opportunistic disease usually prevented by trimethoprim-sulfamethoxazole. A 49-year-old HLA-sensitized male with successful late conversion from tacrolimus-based to belatacept-based immunosuppression developed <i>P. jirovecii</i> pneumonia for which he presented several risks factors: low lymphocyte count with no CD4+ T cells detected since 2 years, hypogammaglobulinemia, history of acute cellular rejection 3 years before, and immunosuppressive treatment (belatacept, everolimus). Because of respiratory gravity in the acute phase, the patient was given oxygen, corticosteroids, and trimethoprim-sulfamethoxazole. Thanks to the improvement of respiratory status, and because of the renal impairment, trimethoprim-sulfamethoxazole was converted to atovaquone for 21 days. Indeed, after 1 week on intensive treatment, the benefit-risk balance favored preserving renal function according to respiratory improvement status. <i>P. jirovecii</i> pneumonia prophylaxis for the next 6 months was monthly aerosol of pentamidine. Long-term safety studies or early/late conversion to belatacept did not report on <i>P. jirovecii</i> pneumonia. Four other cases of <i>P. jirovecii</i> pneumonia under belatacept therapy were previously described in patients having no <i>P. jirovecii</i> pneumonia prophylaxis. Studies on the reintroduction of <i>P. jirovecii</i>pneumonia prophylaxis after conversion to belatacept would be of interest. It could be useful to continue regular evaluation within the second-year post-transplantation regarding immunosuppression: T-cell subsets and immunoglobulin G levels.