Mixed Infections Research Articles

Metagenomic Next-Generation Sequencing in the Diagnosis of Pulmonary Infections after Allogeneic Hematopoietic Stem Cell Transplantation

Early and accurate identification of pathogens in pulmonary infections after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is critically important. The clinical usefulness of metagenomic next-generation sequencing (mNGS) in the diagnosis of pulmonary infections after allo-HSCT remains under discussion. This multicenter retrospective study was conducted to compare mNGS and conventional microbiological tests (CMTs) in identifying the pathogens of pulmonary infections in allo-HSCT recipients. One hundred forty allo-HSCT recipients with suspected pulmonary infections who underwent bronchoscopy were included. mNGS and CMTs performed on bronchoalveolar lavage fluid specimens showed 71.4% positivity on mNGS compared to 55.0% positivity on CMTs. mNGS identified 182 pathogens, including bacteria (n = 88), fungi (n = 35) and viruses (n = 59), compared to 106 pathogens detected by CMTs (bacteria, n = 31; fungi, n = 24; viruses, n = 51). Pulmonary infection was finally diagnosed in 98 patients, including 22 bacterial, 7 fungal, 18 viral, and 48 mixed infections and 3 infections with an unknown pathogen. Mixed infections were identified in 50.5% of the patients with pulmonary infection. The sensitivity of mNGS and CMTs for diagnosing pulmonary infections was 88.8% and 69.4%, respectively (P = .001), and the specificity were 81.0% and 85.7%, respectively (P = .688). Our findings suggest that mNGS may be a promising technology for diagnosing pulmonary infections in allo-HSCT recipients.

Occurrence and identification of lungworms in Iranian wild boars

This study represents the first investigation into the occurrence and identification of Metastrongylus spp. in wild boars (Sus scrofa) in Iran, utilizing both molecular and morphological methods. Thirteen wild boars from Kerman Province were examined, with 92.3% found to be infected with at least one species of Metastrongylus. Mixed infections were observed in 38.46% of the animals. Morphological and molecular analyses confirmed the presence of M. pudendotectus and M. salmi, with prevalence rates of 76.9% and 53.9%, respectively. Histopathological examination revealed transverse and longitudinal sections of Metastrongylus parasites within the airways, causing partial to complete obstruction, interstitial pneumonia, and inflammatory responses. The study also highlights the public health significance of these parasites. The higher prevalence observed compared to earlier studies suggests changes in environmental conditions, host dynamics, or agricultural practices as possible factors, warranting further investigation. The findings underscore the need for comprehensive surveillance and control measures to mitigate the risk of zoonotic transmission, particularly in regions with significant wild and domestic swine populations. This study contributes to the understanding of Metastrongylus spp. distribution and their pathological impact, emphasizing the ecological importance of wild boars and the necessity for continued monitoring and research to prevent and control infections in both animal and human populations.

Clinical diagnostic value of metagenomic next-generation sequencing in patients with acute infection in emergency department

ObjectiveTo explore the value of metagenomic next-generation sequencing (mNGS) and culture in microbial diagnosis of patients with acute infection. MethodsWe retrospectively analyzed 206 specimens from 163 patients who were admitted to the emergency department of The First Affiliated Hospital of Sun Yat-sen University between July 2020, and July 2021. We evaluated the diagnostic efficacy of mNGS and in-hospital traditional culture. ResultsThe total positive rate of mNGS was significantly higher than that culture methods (71.4 % vs 40.8 %, p < 0.001), while the sensitivity and accuracy of mNGS were found to be 92.9 % and 88.2 % respectively. However, culture exhibited superior specificity with a value of 92.6 % compared to 75.9 % for mNGS. The detection efficiency of mNGS and culture for fungi was comparable, but mNGS showed superior performance for bacterial detection. In the analysis of sepsis samples, mNGS outperformed traditional culture methods in diagnosing various types of samples, especially for sputum and bronchoalveolar lavage fluid. Among the identified infections, bacterial infections were the most common single infection (37.5 %). Additionally, bacterial-fungal infections represented the most prevalent form of mixed infection (77.3 %). Candida albicans and Staphylococcus aureus were identified as the predominant pathogens in the survival and death groups, respectively. No significant differences in microbial diversity were observed. ConclusionCompared to culture methods, mNGS demonstrates superior positive rates, sensitivity, and accuracy in the rapid detection of acute infections, particularly in critically ill patients such as those with sepsis. This capability establishes a foundation for the swift and precise identification of pathogens, allowing for the analysis of clinical indicators and patient prognosis based on the extensive data generated from mNGS.

The prevalence and mechanisms of heteroresistance to ceftazidime/avibactam in KPC-producing Klebsiella pneumoniae.

To investigate the prevalence and mechanisms of ceftazidime/avibactam heteroresistance in KPC-producing Klebsiella pneumoniae (KPC-KP) isolates, as well as the role of heteroresistance in the transition of ceftazidime/avibactam susceptibility to resistance. Clinical KPC-KP isolates were obtained from a tertiary hospital in China from 2016 to 2017 and 2019 to 2020. Antimicrobial susceptibility was determined by the broth microdilution method. Population analysis profiles were used to assess ceftazidime/avibactam heteroresistance. WGS and molecular cloning were conducted to reveal heteroresistance mechanisms and molecular characteristics. The findings indicated that the transition of ceftazidime/avibactam susceptibility to resistance during the treatment of KPC-KP infection is primarily attributed to the heteroresistance exhibited by KPC-KP isolates towards ceftazidime/avibactam. Among 355 ceftazidime/avibactam-susceptible KPC-KP isolates (indicating a resistance rate of 0%), 41 (11.55%) exhibited ceftazidime/avibactam heteroresistance, with the primary mechanism being the presence of KPC mutant subpopulations. These KPC variants, arising from point mutations, deletions and insertions, significantly increased ceftazidime/avibactam resistance while alongside enhanced carbapenem susceptibility. Notably, 11 new KPC variants were identified. Furthermore, four heteroresistant isolates were caused by mixed infection involving subpopulations carrying NDM-1 or NDM-5. Phylogenetic analysis indicated that the clonal spread of ST11-KL64 KPC-KP may be correlated with the prevalence of heteroresistance. Ceftazidime/avibactam heteroresistance, primarily driven by pre-existing KPC variants, underscores the importance of considering heteroresistance in ceftazidime/avibactam therapeutics. Awareness of these dynamics is crucial for the effective and sustainable clinical application of ceftazidime/avibactam.

Predictive performance of Metagenomic Next Generation Sequencing in early detection of post-liver transplantation infections

ObjectiveTo evaluate the predictive performance of metagenomic next-generation sequencing (mNGS) in identifying and predicting pulmonary infections following liver transplantation and to investigate its association with patient outcomes within the initial four-week post-transplantation period. MethodsWe retrospectively analyzed 41 liver transplant patients with suspected pulmonary infections from August 2022 to May 2023. Bronchoalveolar lavage fluid (BALF) samples were collected on the first postoperative day for metagenomic next generation sequencing (mNGS) and culture. The predictive capability of mNGS for subsequent infections was assessed by monitoring inflammatory biomarkers and comparing the detection rates with culture methods. Real-time Polymerase Chain Reaction (Rt-PCR) was used to monitor Human betaherpesvirus 5 (CMV) and Human parvovirus B19 (B19) weekly during a four-week postoperative period. Inflammatory biomarkers and blood coagulation function were evaluated on specific days throughout the first, third, fifth, and during four weeks following surgery. The study was conducted until August 2023 to evaluate the patients' prognostic survival outcome, classifying them into groups based on the mortality and survival. ResultsThe analysis included a total of 41 patients, comprising 32 males and 9 females, with an average age of 52 (47, 63) years. Within one week after liver transplantation, there were 7 cases of bacterial infections, 5 cases of fungal infections, 19 cases of mixed infections, 8 cases without any infection, and 2 cases with unidentified pathogen-associated infections. mNGS successfully predicted 39 (72 %) strains of pathogens, while culture-based methods only detected 28 (52 %) strains. Among the 8 patients diagnosed as non-infected, culture methods identified positive results in 4 cases (50 %), whereas mNGS yielded positive results in 7 cases (87.5 %). The detection rates of CMV and B19 by Rt-PCR within 4 weeks after liver transplantation were 61 % and 17 %, respectively (25/41, 7/41) among the patients. During the study period, a total of 9 patients succumbed while 32 patients survived. The death group and the survival group exhibited significant differences in CRP, HGB, and INR levels at specific monitoring time points. The proportion of CMV detection in blood was significantly higher in the death group compared to the surviving group. Elevated CRP level was identified as a prognostic risk factor. ConclusionsDespite the presence of false positives, mNGS still presents a potential advantage in predicting pulmonary infection pathogens following liver transplantation. Furthermore, the levels of CRP and CMV carrier status within four weeks post-surgery exhibit significant associations with patient survival and prognosis.

Effect of micronutrient deficiency on protozoal infection in stunting toddler

&lt;p class="pf0"&gt;&lt;strong&gt;&lt;em&gt;Background: &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;Stunting is a developmental disorder caused by chronic inadequate nutritional intake in toddlers, leading to deficiencies in various nutrients, including iron and zinc. This can weaken toddlers' immunity, making them more vulnerable to infectious diseases, such as intestinal protozoa. An infected digestive tract can interfere with absorbing nutrients and reduce appetite, causing the infection to persist and become chronic. Chronic infection triggers the formation of pro-inflammatory cytokines that affect bone growth, hindering growth and causing stunting.&lt;/em&gt;&lt;em&gt;&lt;/em&gt;&lt;/p&gt;&lt;p class="pf0"&gt;&lt;strong&gt;&lt;em&gt;Objectives: &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;This study confidently investigates the correlation between iron and zinc micronutrient intake and intestinal protozoan infection in stunting toddlers in Jember District.&lt;/em&gt;&lt;em&gt;&lt;/em&gt;&lt;/p&gt;&lt;p class="pf0"&gt;&lt;strong&gt;&lt;em&gt;Methods: &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;The research is observational and cross-sectional, with 568 stunting toddlers aged 0-59 months in Jember District, East Java as study subjects. Iron and zinc intake were collected through 2x24-hour food-recall questionnaire interview and processed using the Nutrisurvey software. Protozoan infection status in toddler feces samples was identified using direct smear and modified Ziehl-Neelsen methods. The data analysis method chosen was the Spearman correlation test.&lt;/em&gt;&lt;/p&gt;&lt;p class="pf0"&gt;&lt;strong&gt;&lt;em&gt;Results: &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;The study found that stunted toddlers in Jember District had mostly deficient micronutrient intake. The average iron intake is 2.56±3.06 mg and the average zinc intake is 1.8±1.47 mg. Protozoan infection was found in this study has a prevalence of 15.7%, with identified species including Entamoeba histolytica (3.3%), Giardia lamblia (2.6%), Cryptosporidium parvum (3.2%), Blastocystis hominis (5.3%), and mixed infections (1.2%). The statistical analysis confirms that there is no significant correlation between the intake of micronutrients iron (p=0.91;r=0.005) and zinc (p=0.76;r=0.013) and intestinal protozoan infection in stunting toddlers in Jember District.&lt;/em&gt;&lt;/p&gt;&lt;p class="pf0"&gt;&lt;strong&gt;&lt;em&gt;Conclusions: &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;Based on the data, protozoan infection may be caused by multiple factors, such as parenting practices and family socioeconomics. However, improving nutrition by education and giving additional supplementation are crucial to reduce the prevalence of malnutrition in toddlers.&lt;/em&gt;&lt;em&gt;&lt;/em&gt;&lt;/p&gt;

Clinical and Etiology Presentations of Keratitis Caused by Mixed Infectious Agents (Bacteria and Fungi)

Introduction: Microbiological studies following the culture of corneal infiltrates are the gold standard for determining the etiology of infectious keratitis caused by bacteria or fungi; however, even if a culture of the corneal ulcer is obtained. Purpose: To investigate the predisposing factors and clinical and Etiology presentations of keratitis caused by mixed infectious agents (bacteria and fungi). Methods: This was a prospective study of cases with mixed bacterial and fungal keratitis, done between July to December 2022 in Department of Ophthalmology, Netraloy Eye Care Center, Thanthania, Bogura Sadar, Bogura, Bangladesh. Sixty (60) cases of mixed bacterial and fungal keratitis were identified. Samples (corneal swabs and scrapings) were collected aseptically from corneal ulcer patients. Isolation and identification of the microbial agents and antimicrobial susceptibility testing were done in the Microbiology and Pathology departments. Results: 38 cases (63.3%) were men, and the mean age was 54.2±9.3 years. The affected people were mostly (50; 83.3%) residing in the rural areas. 34 patients (56.7%) were involved in agricultural activities. The people of 41- 60 years of age were particularly prone to this disease (40; 66.7%). The most common predisposing factor for mixed keratitis was a history of ocular trauma (42; 70%) and 28 patients (46.7%) had a history of trauma with vegetative matter. The incidence of the disease was highest in the monsoon season, between June to September (32; 53.3%). The most common causative bacterial organisms was Staphylococcus aureus (26;43.3%) followed by Pseudomonas secies (10;16.7%) and among fungal organisms was Aspervgillus fumigates (22;36.7%) followed by Fusarium species (12;20%). Conclusion: The identification of the etiology and the predisposing factors of corneal ulcers in this region are important for the prevention and early treatment of the disease. Usually, patients with mixed bacterial and fungal keratitis have poor prognosis. Thus, when the infectious keratitis is running an atypical course or found unresponsive to the initial medical treatment, the possibility of a mixed infection by bacterial and fungal organisms should be considered.

Bridging Diagnostic Gaps: Utilising HiCrome Agar and Tetrazolium Reduction Medium for the Rapid and Presumptive Identification and Speciation of Candida Species in Vulvovaginal Candidiasis in Low-Resource Environments.

Background Vulvovaginal candidiasis (VVC) is a common fungal infection caused by an overgrowth of Candida species, primarily Candida albicans (C. albicans). Using HiCrome agar and tetrazolium reduction medium offers cost-effectiveness in Candida detection by eliminating the need for additional tests, reducing equipment costs compared to automated systems, and simplifying workflow with direct species identification while maintaining high specificity. They expedite detection by directly identifying Candida species based on colony colour, bypassing the multiple steps of phenotypic methods. This efficiency saves time in the laboratory, providing rapid results without the extended processing times associated with automated systems and facilitating prompt diagnosis and treatment decisions. These diagnostic tools are especially valuable in low-resource environments where a quick and accurate diagnosis of VVC is crucial for effective treatment and management of antifungal resistance. Aims and objectives This study aims to evaluate the efficacy of HiCrome agar and tetrazolium reduction medium's efficacy in speciating Candida species in VVC cases. Materials and methods A cross-sectional observational study was conducted at Saveetha Medical College and Hospitals, Chennai, India, over six months. High vaginal swabs from 126 patients suspected of VVC were collected and plated on Sabouraud dextrose agar (SDA), HiCrome Candida differential agar (Himedia, Mumbai, India), and tetrazolium reduction medium. The results were compared with those obtained from the VITEK2 compact system (bioMérieux, Marcy-l'Étoile, France). Results Of the 126 samples, 74.6% showed single yeast infections, 7.9% displayed mixed yeast infections, and 17.5% showed no growth. A total of 114 Candida isolates were identified. Both HiCrome agar and tetrazolium reduction medium accurately identified all isolates, with complete concordance with the VITEK2 compact system. The most commonly isolated species were C. albicans (55.2%), Candida tropicalis (32.4%), Candida glabrata (8.8%), and Candida parapsilosis (3.6%). Both media provided rapid and accurate presumptive identification in low-resource settings. Conclusions HiCrome agar and tetrazolium reduction medium demonstrated high sensitivity and specificity in identifying Candida species. These methods are reliable for rapid and accurate diagnosis, particularly in resource-limited settings. However, they may require supplementary tests for definitive species identification. The adoption of these diagnostic tools represents a significant advancement in clinical microbiology, improving VVC management and addressing antifungal resistance.

Investigation of Tick-Borne Pathogens in Ixodes Ticks from Bosnia and Herzegovina.

Limited information is available regarding the presence of tick-borne pathogens and their distribution within Ixodes species in Bosnia and Herzegovina. This study aimed to identify Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato (s.l.) in Ixodes ticks collected from domestic and wild animals and vegetation in different regions across Bosnia and Herzegovina. A total of 7438 adult ticks, including 4526 Ixodes ricinus, Ixodes canisuga, and Ixodes hexagonus, were collected. Real-time PCR screening of 450 pooled I. ricinus samples revealed a 22.1% infection rate with at least one pathogen. Rickettsia spp. (6.3%) were found in ticks from dogs, cats, and goats, Babesia spp. (3.1%) in ticks from dogs and cattle, A. phagocytophilum (8.8%) in ticks from dogs, goats, and cattle, and B. burgdorferi s.l. (3.4%) in ticks from dogs and cats. Mixed infections with B. burgdorferi s.l. and A. phagocytophilum, as well as B. burgdorferi s.l. and Rickettsia spp., were found in two pools of I. ricinus from dogs and cats, respectively. Additionally, co-infection with Rickettsia spp. and A. phagocytophilum was confirmed in three tick pools from dogs and goats. Each tick from these pooled samples was individually retested to confirm the presence of pathogens. In the examined pooled samples of I. canisuga (1) and I. hexagonus (6), none of the tested pathogens were detected. Our findings represent the first detection of Rickettsia spp., Babesia spp., A. phagocytophilum, and B. burgdorferi s.l. in I. ricinus collected from domestic animals and vegetation in Bosnia and Herzegovina. Considering the established infection rates, the detection of tick-borne pathogens in adult ticks collected from domestic animals and vegetation enriches the current knowledge of the presence of tick-borne pathogens at the local, regional, national, and broader levels.

Serotype-specific clinical features and spatial distribution of dengue in northern Kerala, India.

Collection and compilation of spatial, meteorological, entomological, and virological data are critical in mitigating climate-sensitive emerging infections like dengue. This study was a holistic attempt to understand the dengue situation in the Kasaragod district of Kerala, India. This cross-sectional study was conducted in 13 health institutions from June to July 2021. Adult patients presenting with fever and testing positive for NS1 ELISA were subjected to Dengue RT-PCR and serotyping. The spatial and clinical features of the RT-PCR-positive patients, the district's meteorological data, and the vector indices were studied. The pre-epidemic months were marked by intermittent rainfall, peak ambient temperature and high larval indices. Among the 136 dengue RT-PCR patients studied, 41.2% had DENV2 followed by DENV1 (22.8%), DENV3 (5.9%) and DENV4 (4.4%); with 25% mixed infections. DENV1 showed a higher risk of gastrointestinal manifestations (80.6%, p=0.019) and musculoskeletal symptoms (77.4%, p=0.026) compared with other serotypes. In the context of dengue hyperendemicity, the possibility of an emerging serotype's dominance coupled with the mixing up of strains should warn the health system regarding future outbreaks. Furthermore, the study emphasizes the importance of monitoring larval indices and the window of opportunity to intervene between environmental predictors and dengue outbreaks.

Rhinocerebral mucormycosis: A clinicopathological analysis of COVID-19-associated mucormycosis.

During the coronavirus disease 2019 (COVD-19) pandemic, the incidence of mucormycosis also increased, especially affecting individuals who have had the COVID-19 infection in the past. The aim of the study is to assess risk factors and clinical and histopathological features of mucormycosis in post-COVID-19 cases. This is a retrospective study conducted in a tertiary care COVID-19-dedicated hospital, Dehradun, Uttarakhand, India, over a period of 2 months during the COVID-19 pandemic. All surgical specimens submitted for histopathology with a suspected diagnosis of mucormycosis were included. Histopathology was considered the gold standard. All histopathologically confirmed cases were studied in detail with respect to histopathology, clinico-radiological features, and microbiological results. Of 25 cases with clinical diagnosis of mucormycosis, nine were histopathologically confirmed as mucormycosis. Seven patients had diabetes, while two did not have any co-morbidity. The fungal load was heavy in 50% cases, and the proportion of necrosis was higher with diabetes mellitus, as compared to non-diabetic and non-co-morbidity patients. Angioinvasion (33.3% cases), soft-tissue invasion (44.4%), Splendor-Hoeppli phenomenon (44.4%), and neural invasion (11.1%) were also present. Mixed infection (Mucormycosis and Aspergillus species) was present in three of the cases who also had diabetes. The microbiological investigations were positive in only 55.5% cases. Post-COVID Mucormycosis has fatal outcomes. Early diagnosis and treatment are the key to successful treatment. Early and reliable diagnosis can be offered by histopathological examination.

A metagenomic next-generation sequencing (mNGS)-based analysis of bronchoalveolar lavage samples in patients with an acute exacerbation of chronic obstructive pulmonary disease.

The role of the bronchoalveolar lavage fluid (BALF) microbiome in acute exacerbations of chronic obstructive pulmonary disease (AECOPD) remains unclear. The advent of the metagenomic next-generation sequencing (mNGS) has made it possible to reveal the complex microbiome composition of the respiratory tract. This study aimed to explore whether there are differences in the BALF microbiome of AECOPD patients with different lung functions. We enrolled 55 AECOPD patients and divided them into a mild group (n = 31) and a severe group (n = 24) according to their lung function. We collected BALF and submitted it to mNGS and bioinformatics analysis. At the species level, mNGS identified 264 bacteria, 13 fungi and 12 viruses in the mild group, and 174 bacteria, 6 fungi and 6 viruses in the severe group. Mixed bacterial and viral infection occurred in both groups. At the genus level, Rothia and Veillonella were more abundant in the mild group, while Pseudomonas and Staphylococcus were more abundant in the severe group. At the species level, compared with the mild group, the relative abundance of Haemophilus influenzae and Pseudomonas aeruginosa was increased in the severe group. Besides, the BALF microbiome composition was similar between the two groups, and there was no significant difference in α and β diversity. Forced expiratory volume in 1s/forced vital capacity (FEV1/FVC) (%) showed no significant correlation with the Shannon or Simpson index. The microbiome abundance was different between the mild and severe groups; however, microbiome diversity was similar between the two groups. Based on our findings, Haemophilus influenzae and Pseudomonas aeruginosa may be the pathogenic bacteria that cause the difference in lung function in patients with AECOPD.

Clinical impact of metagenomic next-generation sequencing of bronchoalveolar lavage fluids for the diagnosis of pulmonary infections in respiratory intensive care unit

BackgroundThe real-world clinical impact of mNGS on BALF in the respiratory intensive care unit (RICU) is not yet fully understood. MethodsWe investigated the clinical impact of mNGS on BALF samples obtained from 92 patients admitted to the RICU over a 2-year period. We utilized both mNGS and culture methods to evaluate the effectiveness of mNGS in diagnosing pulmonary infections. The clinical impact of mNGS were evaluated by the clinician committees. ResultsAmong the 92 diagnosed patients, 78 cases (84.7 %) were determined to have infectious diseases caused by pathogenic microorganisms, and the bacterial infections constituted the most prevalent diagnostic category. For mixed infection, the most common type was the Pneumocystis jironecii and cytomegalovirus co-infection. The mNGS results had a positive impact on the clinical management of 43 cases (46.7 %). Moreover, 19 cases (44.2 %) of positive clinical impacts were solely based on new diagnoses made possible by mNGS results. These new diagnoses were particularly helpful for identifying rare pathogens, which could not be detected by conventional diagnostic methods. ConclusionsThe BALF mNGS has a positive real-world impact in RICU. Clinician committee play a critical role in ensuring the appropriate use of mNGS.

Single and mixed infections of six major potato viruses in four major potato-growing districts of eastern Ethiopia

Potato (Solanum tuberosum L.) is a co-staple food and a source of cash income for the smallholder farmers' in eastern Ethiopia; however, the productivity of the crop is constrained by biotic and abiotic factors. Of which viruses are considered to be major yield limiting factor, but information is lacking on the prevalence and distribution of potato viruses in major growing districts of eastern Ethiopia. This research is therefore initiated to assess and determine the status of six major and multiple potato viruses' infection at farmers' fields during the main rainy and irrigation seasons in four major potato-growing districts of eastern Ethiopia. The dot-blot assays performed for 1000 leaf samples collected from 20 farmers' fields during the two seasons indicated an overall high incidence (71.9% samples detected positive for at least one virus) of potato virus in the surveyed districts of the regions. The most prevalent (19.4%) type of infection was the mixture of all six potato viruses. Among the major viruses, the results also confirmed PVM as the most prevalent (57.3%) virus followed by PVY (48.1%), whereas PVA (45.1%), which was not commonly detected in the other regions of the country, was detected as third most prevalent virus in the current study. The comparison of the results of virus detection in the samples from two seasons indicated remarkable differences in the incidence and prevalence of viruses, where an increasing trend of virus incidence and mixed virus infections from 59 to 84.8% and 41.2 to 72%, respectively, were recorded from rainy to irrigation season. The two viruses; PVX and PVS, which were identified as least prevalent during rainy season, were ranked the 3rd and 4th most prevalent viruses after PVM and PVY during irrigation season. The detected high incidence of virus in potato plants during this first systemic research conducted suggested the importance of continuing the intensive survey and detecting the status of viruses' infection in potato growing districts of eastern Ethiopia. It is also suggested to create awareness among smallholder farmers about the management practices to reduce the effects of viruses’ infection on the productivity of potato in eastern Ethiopia.

Mixed Wolbachia infections resolve rapidly during in vitro evolution.

The intracellular symbiont Wolbachia pipientis evolved after the divergence of arthropods and nematodes, but it reached high prevalence in many of these taxa through its abilities to infect new hosts and their germlines. Some strains exhibit long-term patterns of co-evolution with their hosts, while other strains are capable of switching hosts. This makes strain selection an important factor in symbiont-based biological control. However, little is known about the ecological and evolutionary interactions that occur when a promiscuous strain colonizes an infected host. Here, we study what occurs when two strains come into contact in host cells following horizontal transmission and infection. We focus on the faithful wMel strain from Drosophila melanogaster and the promiscuous wRi strain from Drosophila simulans using an in vitro cell culture system with multiple host cell types and combinatorial infection states. Mixing D. melanogaster cell lines stably infected with wMel and wRi revealed that wMel outcompetes wRi quickly and reproducibly. Furthermore, wMel was able to competitively exclude wRi even from minuscule starting quantities, indicating that this is a nearly deterministic outcome, independent of the starting infection frequency. This competitive advantage was not exclusive to wMel's native D. melanogaster cell background, as wMel also outgrew wRi in D. simulans cells. Overall, wRi is less adept at in vitro growth and survival than wMel and its in vivo state, revealing differences between the two strains in cellular and humoral regulation. These attributes may underlie the observed low rate of mixed infections in nature and the relatively rare rate of host-switching in most strains. Our in vitro experimental framework for estimating cellular growth dynamics of Wolbachia strains in different host species and cell types provides the first strategy for parameterizing endosymbiont and host cell biology at high resolution. This toolset will be crucial to our application of these bacteria as biological control agents in novel hosts and ecosystems.

ЦИРКОВИРУС-МИКОПЛАЗМЕННАЯ ИНФЕКЦИЯ ПОРОСЯТ В ЭКСПЕРИМЕНТЕ

The aim of the study is to investigate the manifestation of circovirus-mycoplasma infection in piglets both as a mixed infection and as a monoinfection in the experiment. Objectives: to study the interaction of circovirus type 2 with Mycoplasma hyopneumoniae in the body of weaned piglets, as well as the manifestation of monoinfection in piglets during experimental infection; to assess the pathogenicity of PCV2d/M.hyo mixed infection, PCV2d monoinfection and M. hyo monoinfection; to identify the clinical and pathomorphological manifestation of mixed and single circovirus and mycoplasma infection in the experiment. Four groups of 23-day-old piglets (5 heads each) were formed and tested for the absence of pathogens. Experimental group I – mixed circovirus-mycoplasma infection, experimental group II – PCV2d monoinfection, experimental group III – M.hyo monoinfection, group IV – control. After infection, rectal temperature was measured daily and clinical manifestations were observed, nasal swabs were taken to determine the number of copies of M. hyo nucleic acid, blood was taken from each piglet weekly to detect viral nucleic acid and antibodies. All experimental and control piglets were euthanized on the 35th day from the beginning of the experiment. Pathomorphological and pathohistological studies were carried out. The experimental study reveals the synergistic pathogenicity of PCV2d and M. hyo in mixed infection. In experimental infection of weaning piglets with both M. hyo monoinfection and M. hyo/PCV2d mixed infection, it was found that M. hyo nucleic acid was detected in nasal smears in 80 % of animals and in lung tissue in 100 %. The number of copies of the PCV2d circovirus genome in the inguinal lymph node tissue of piglets in the mixed infection group was higher than in the monoinfection group. Circovirus type 2 in piglets can proceed latently, and circovirus-mycoplasma infection can be a complex respiratory disease.

Epidemiology, Clinical Manifestations, Treatment, and Outcome of Mucormycosis: A Review of 77 Cases From a Single Center in France.

The aim of this study was to assess the epidemiology, clinical manifestations, and outcome of mucormycosis over 15 years in a single center in France. We conducted a retrospective analysis of all mucormycosis cases in our institution from 1 January 2006 to 31 December 2020 and analyzed patients' medical records, laboratory results, and treatment to describe the epidemiology, clinical manifestations, diagnosis, treatment, and outcome. Mucorales quantitative polymerase chain reaction (qPCR) for the diagnosis was implemented in 2015. Seventy-seven mucormycosis cases were analyzed in 77 patients, with a median age of 54 years (60% male). Identified risk factors were hematological diseases (46 cases [60%]), solid malignancies (2 cases), solid organ transplants (3), burns (18), diabetes only (7), and trauma (1). Sites of infection were lungs (42%), sinus (36%), skin (31%), central nervous system (9%), liver (8%), others (6%), and disseminated (12%). Diagnosis remained difficult and qPCR contributed to mucormycosis diagnosis in 30% of cases. Among hematology patients, serum qPCR was the only positive test in 15% of cases. A mixed mold infection was diagnosed in 24 of 77 (31%) patients. Surgical treatment was undertaken in 43 (56%) cases. Most patients received liposomal amphotericin B (89%), with a combination therapy in 18 of 77 cases (23%). Three-month survival rate was 40% (95% confidence interval [CI], .30-.53]). As for treatment, adjunction of surgery (hazard ratio, 0.47 [95%CI, .25-.91); P = 0.02) was associated with lower mortality. Mucormycosis remained associated with high mortality, especially in the hematological and burn populations. Surgery in combination with antifungal treatment was associated with improved survival.

The plasma viral communities associate with clinical profiles in a large-scale haematological patients cohort

BackgroundHaematological patients exhibit immune system abnormalities that make them susceptible to viral infections. Understanding the relationship between the virome in the blood plasma of haematological patients and their clinical characteristic is crucial for disease management. We aimed to explore the presence of viral pathogens and identify close associations between viral infections and various clinical features.ResultsA total of 21 DNA viruses and 6 RNA viruses from 12 virus families were identified from 1383 patients. Patients with haematological diseases exhibited significantly higher diversity, prevalence, and co-detection rates of viral pathogens. During fever episodes, pathogen detection was notably higher, with Epstein-Barr virus (EBV) and Mucorales infections being the most probable culprits for fever symptoms in non-haematological patients. The detection rate of torque teno virus (TTV) significantly increases in haematological patients after transplantation and during primary lung infections. Additionally, TTV-positive patients demonstrate significantly higher absolute neutrophil counts, while C-reactive protein and procalcitonin levels are notably lower. Furthermore, TTV, cytomegalovirus, and parvovirus B19 (B19V) were found to be more prevalent in non-neutropenic patients, while non-viral pathogenic infections, such as Gram-negative bacteria and Mucorales, were more common in neutropenic patients. Pegivirus C (HPgV-C) infection often occurred post-transplantation, regardless of neutropenia. Additionally, some viruses such as TTV, B19V, EBV, and HPgV-C showed preferences for age and seasonal infections.ConclusionsAnalysis of the plasma virome revealed the susceptibility of haematological patients to plasma viral infections at specific disease stages, along with the occurrence of mixed infections with non-viral pathogens. Close associations were observed between the plasma virome and various clinical characteristics, as well as clinical detection parameters. Understanding plasma virome aids in auxiliary clinical diagnosis and treatment, enabling early prevention to reduce infection rates in patients and improve their quality of life.EnXnzQJ5tBc4iYM41USpCeVideo

Clinical efficacy of a parasiticide formulation containing eprinomectin, esafoxolaner and praziquantel (NexGard® Combo) in the treatment of natural feline aelurostrongylosis and troglostrongylosis

The metastrongyloids Aelurostrongylus abstrusus and Troglostrongylus brevior are primary causes of feline clinical respiratory disease. The present field trial evaluated the clinical efficacy of a spot-on formulation containing eprinomectin, esafoxolaner and praziquantel (NexGard® Combo) administered per label recommendations to cats affected with aelurostrongylosis and/or troglostrongylosis. Overall, 36 naturally infected cats were randomly assigned to Group 1 (G1) or Group 2 (G2) of 18 cats each. The two groups included 6 cats with A. abstrusus, T. brevior and mixed infection, each. All cats completed the study. Cats in G1 were treated on study Days (SDs) 0 and 28±2. Cats in G2 served as negative control until SD 56±2 and were then treated on SD 56±2 and 84±2. On SD 0/-7, 28±2 and SD 56±2 all cats were subjected to parasitological (quali-quantitative Baermann) and clinical examinations (physical exams and thoracic X-rays). Hematology and biochemistry analyses were performed on SD 0/-7 and SD 56±2. On SD 84±2 quali-quantitative Baermann, clinical examination and thorax radiography were performed on all G2 cats and on two G1 cats that still had radiographic alterations on SD 56±2. On SD 112±2 all G2 cats were subjected to parasitological and clinical evaluations and one cat from G1 that still had radiographic signs at SD 84±2 was clinically and radiographically evaluated. Efficacy criteria were the reduction of larval shedding in faeces and the clinical response in terms of pathological and radiographic scores after treatment compared to the baseline. An efficacy of 100 % based on LPG reduction was recorded after one (20/24 cats) or two (all 24 cats) treatments in cats with single infection by A. abstrusus or T. brevior. For cats with mixed infections, larval shedding was stopped after one (11/12 cats) or two (all 12 cats) treatments. Statistically significant clinical and radiographic improvement was evident in all study cats after 2 treatments. The present data show that two monthly treatments with NexGard® Combo stopped larval shedding and led to a significant clinical recovery and a complete resolution of radiographic abnormalities in cats infected with A. abstrusus and/or T. brevior.

Soluble expression and immunogenicity analysis of capsid proteins of porcine circoviruses types 2, 3, and 4

Porcine circoviruses (PCVs) contain four types: PCV1, PCV2, PCV3, and PCV4, all of which can infect pigs. Among them, PCV1 is non-pathogenic, and PCV2 can cause porcine circovirus diseases (PCVD) or porcine circovirus-associated diseases (PCVAD). Although the pathogenicity of PCV3 and PCV4 is still controversial, increasing evidence shows that PCV3 and PCV4 can cause PCV-related disease. However, mixed infection of PCV2, PCV3, and PCV4 with other pathogens often occurs in large-scale pig breeding, bringing severe economic losses to the global pig industry. In this study, the soluble recombinant proteins of PCV2, PCV3, and PCV4 Cap were expressed by the prokaryotic expression system and biotinylated to combine with the Streptavidin magnetic beads, followed by immunogenicity evaluation of the recombinant proteins. Furthermore, we also assessed the efficacy and immunogenicity of trivalent recombinant proteins conjugated with different adjuvants in mice. The results showed that the highly effective anti-PCV serum was successfully prepared, and the recombinant proteins conjugated with different adjuvants produced various degrees of humoral and cellular immunity in mice. Three recombinant proteins are effective immunogens, and the trivalent proteins coupled with the aluminum adjuvant or GM-CSF-CpG for two-dose immunization can stimulate prominent humoral and cellular immunity against PCVs in vivo. The soluble recombinant proteins are the most promising candidate for developing a trivalent vaccine against PCVs (PCV2, PCV3, and PCV4) infection simultaneously.