Dual Infection Research Articles

The Enemy of my Enemy is my Friend: Immune-Mediated Facilitation Contributes to Fitness of Co-Infecting Helminths.

The conceptual understanding of immune-mediated interactions between parasites is rooted in the theory of community ecology. One of the limitations of this approach is that most of the theory and empirical evidence has focused on resource or immune-mediated competition between parasites and yet there is ample evidence of positive interactions that could be generated by immune-mediated facilitation. We developed an immuno-epidemiological model and applied it to longitudinal data of two gastrointestinal helminths in two rabbit populations to investigate, through model testing, how immune-mediated mechanisms of parasite regulation could explain the higher intensities of both helminths in rabbits with dual than single infections. The model framework was selected and calibrated on rabbit population A and then validated on the nearby rabbit population B to confirm the consistency of the findings and the generality of the mechanisms. Simulations suggested that the higher intensities in rabbits with dual infections could be explained by a weakened or low species-specific IgA response and an asymmetrical IgA cross-reaction. Simulations also indicated that rabbits with dual infections shed more free-living stages that survived for longer in the environment, implying greater transmission than stages from hosts with single infections. Temperature and humidity selectively affected the free-living stages of the two helminths. These patterns were comparable in the two rabbit populations and support the hypothesis that immune-mediated facilitation can contribute to greater parasite fitness and local persistence.

High-Throughput Sequencing Reveals Tobacco and Tomato Ringspot Viruses in Pawpaw.

Pawpaw (Asimina triloba) trees exhibiting stunting and foliar mosaic, chlorosis, or distortions were observed in New York. In 2021, leaf samples from two symptomatic trees and a sapling, as well as two asymptomatic trees, were tested for the presence of viruses and viroids by high-throughput sequencing (HTS) using total RNA after ribosomal RNA depletion. HTS sequence information revealed tobacco ringspot virus (TRSV) and tomato ringspot virus (ToRSV) in symptomatic but not in asymptomatic leaves. HTS reads and de novo-assembled contigs covering the genomes of both viruses were obtained, with a higher average read depth for RNA2 than RNA1. The occurrence of TRSV and ToRSV was confirmed in the original leaf samples used for HTS and 12 additional trees and saplings from New York and Maryland in 2022 by RT-PCR combined with Sanger sequencing, and DAS-ELISA. Single infections by TRSV in 11 of 14 trees and dual infections by TRSV and ToRSV in 3 of 14 trees were identified. The nucleotide sequence identity of partial gene fragments of TRSV and ToRSV was high among pawpaw isolates (94.9-100% and 91.8-100%, respectively) and between pawpaw isolates and isolates from other horticultural crops (93.6-100% and 71.3-99.3%, respectively). This study is the first to determine the virome of pawpaw.

Evidence of orthohantavirus and leptospira infections in small mammals in an endemic area of Gampaha district in Sri Lanka

BackgroundOrthohantaviruses and leptospira are emerging zoonotic pathogens of high public health significance. The epidemiology of orthohantavirus infections and leptospirosis is similar and presents related clinical pictures in humans. However, a paucity of data on actual reservoir hosts for orthohantaviruses and leptospira exists. Therefore, this study aimed at determining the occurrence of orthohantaviruses and leptospira in small mammals captured in an endemic region of Sri Lanka.MethodsRodents and shrews were morphologically and/or genetically identified using morphological keys and DNA barcoding techniques targeting the cytochrome oxidase b subunit gene (Cytb). Lung tissues and sera were subsequently analyzed for the presence of orthohantavirus RNA using qRT-PCR. Sera of rats were tested for IgG antibodies against orthohantaviruses and leptospira.ResultsForty-three (43) small mammals representing: Rattus (R.) rattus (black rat) or R. tanezumi (Asian rat), Suncus murinus (Asian house shrew), R. norvegicus (brown rat) and Mus musculus (house mouse) were investigated. No orthohantavirus RNA was detected from the lung tissue or serum samples of these animals. Elevated levels of IgG antibodies against Puumala orthohantavirus (PUUV) and/or Seoul orthohantavirus (SEOV) antigens were detected in sera of 28 (72%) out of the 39 rats analysed. Interestingly, 36 (92%) of the 39 rats also showed presence of anti leptospira-IgG antibodies in their serum, representing dual infection or dual exposure in 26/39 (66.7%) of examined rats.ConclusionsThis project targets important public health questions concerning the occupational risk of orthohantavirus infections and/or leptospirosis in an endemic region of Sri Lanka. Most rats (72%) in our study displayed antibodies reacting to orthohantavirus NP antigens, related to PUUV and/or SEOV. No correlation between the orthohantavirus and leptospira IgG antibody levels were noticed. Finally, a combination of both morphological and DNA barcoding approaches revealed that several species of rats may play a role in the maintenance and transmission of orthohantavirus and leptospira in Sri Lanka.

Blood Mucorales PCR to track down Aspergillus and Mucorales co-infections in at-risk hematology patients: A case-control study.

Serum Mucorales PCR can precede the final diagnosis of invasive mucormycosis by several days or weeks and could therefore be useful as a non-invasive screening tool. We assessed the performance of a commercial Mucorales PCR assay (MucorGenius®, PathoNostics, Maastricht, The Netherlands) on prospectively collected banked sera from hematology patients at risk for invasive mould infections. We evaluated if there is an underestimated incidence of missed Mucorales co-infections in patients with invasive aspergillosis (IA). We tested Mucorales PCR on the sera of all patients with a diagnosis of at least possible IA (EORTC-MSGERC consensus criteria) before the start of any antifungal therapy, and in a control group of similar high-risk hematology patients without IA (in a 1:4 ratio). When a positive Mucorales PCR was observed, at least 5 serum samples taken before and after the positive one were selected. Mucorales PCR was performed in 46 diagnostic serum samples of cases and in 184 controls. Serum Mucorales PCR was positive in 4 cases of IA (8.7%; 12.9% of probable cases) and in 1 control case (0.5%) (p=0.0061, OR=17.43 (1.90-159.96). Post-mortem cultures of the positive control became positive for Rhizopus arrhizus. Mortality of IA cases with and without a positive Mucorales PCR was not significantly different. Only in the PCR positive control case, serial serum samples before and after the diagnostic sample were also positive. It is not entirely clear what a positive Mucorales PCR in these cases implies since the 4 Mucorales PCR positive cases were treated with antifungals with activity against Mucorales. In addition, PCR was positive only once. This study does not provide enough evidence to implement Mucorales PCR screening. However, our findings emphasize once more the importance of considering the possibility of dual mould infections, even in patients with a positive galactomannan detection.

Cervical Epidural Electrical Stimulation Increases Respiratory Activity through Somatostatin-Expressing Neurons in the Dorsal Cervical Spinal Cord in Rats.

We tested the hypothesis that dorsal cervical epidural electrical stimulation (CEES) increases respiratory activity in male and female anesthetized rats. Respiratory frequency and minute ventilation were significantly increased when CEES was applied dorsally to the C2-C6 region of the cervical spinal cord. By injecting pseudorabies virus into the diaphragm and using c-Fos activity to identify neurons activated during CEES, we found neurons in the dorsal horn of the cervical spinal cord in which c-Fos and pseudorabies were co-localized, and these neurons expressed somatostatin (SST). Using dual viral infection to express the inhibitory Designer Receptors Exclusively Activated by Designer Drugs (DREADD), hM4D(Gi), selectively in SST-positive cells, we inhibited SST-expressing neurons by administering Clozapine N-oxide (CNO). During CNO-mediated inhibition of SST-expressing cervical spinal neurons, the respiratory excitation elicited by CEES was diminished. Thus, dorsal cervical epidural stimulation activated SST-expressing neurons in the cervical spinal cord, likely interneurons, that communicated with the respiratory pattern generating network to effect changes in ventilation.SIGNIFICANCE STATEMENT A network of pontomedullary neurons within the brainstem generates respiratory behaviors that are susceptible to modulation by a variety of inputs; spinal sensory and motor circuits modulate and adapt this output to meet the demands placed on the respiratory system. We explored dorsal cervical epidural electrical stimulation (CEES) excitation of spinal circuits to increase ventilation in rats. We identified dorsal somatostatin (SST)-expressing neurons in the cervical spinal cord that were activated (c-Fos-positive) by CEES. CEES no longer stimulated ventilation during inhibition of SST-expressing spinal neuronal activity, thereby demonstrating that spinal SST neurons participate in the activation of respiratory circuits affected by CEES. This work establishes a mechanistic foundation to repurpose a clinically accessible neuromodulatory therapy to activate respiratory circuits and stimulate ventilation.

Recent cases of co-infection of scrub typhus and leptospirosis: A short clinical study

Scrub typhus and leptospirosis are bacterial zoonotic diseases causing high morbidity and mortality. Both are important causes of acute febrile illness in India. The common epidemiology of both the diseases creates an opportunity of dual infections with these diseases. The present study was conducted at RG Kar Medical College and Hospital to determine simultaneous infection of leptospirosis and scrub typhus in sera of febrile patients and evaluate the cases of serological dual infections. IgM antibodies to both leptospira and scrub typhus were detected by Enzyme linked immunosorbant assay (ELISA). During the study period (August 2022-October 2022), serum samples of patients were received for serology for scrub typhus. Out of these samples, 5 patients diagnosed for scrub typhus infection also had IgM antibodies to leptospira, thus indicating co-infection of both diseases. The current study adds further to few previously published reports on co-infection of scrub typhus with leptospirosis. As both leptospirosis and scrub typhus present with similar clinical features, co-infection of these two diseases is not uncommon and requires further confirmation at molecular level. Accordingly, cases of dual infections must be considered seriously and choice of therapy should include those drugs that cover for both the infections.

HIV-TB Cross-referral and Collaborative Strategy: 8 Years of Our Experience from An Urban Health Centre in North India

Human immunodeficiency virus (HIV) and tuberculosis (TB) epidemics continue to fuel each other and with dual infections with these two deadly diseases on the rise, it becomes imperative to devise effective HIV-TB collaborative strategies. The present study was designed to evaluate the existing HIV-TB cross-referral mechanism at an urban health centre; to determine HIV sero-prevalence among pulmonary TB patients referred from chest clinic to the integrated counselling and testing centre (ICTC); and to evaluate the TB suspects referred from ICTC to the chest clinic for a possible TB aetiology. The present study was a retrospective analysis of HIV-TB cross-referrals whereby a line list of all the patients referred under this strategy from January 2006 to December 2013 was retrieved and analysed. A total of 3726 TB cases were referred to the ICTC and 641 TB suspects were identified by ICTC counsellors and referred to the chest clinic during this period. HIV sero-prevalence among TB patients was 2.8% (106 of 3726) and TB prevalence among HIV sero-positive and sero-negative TB suspects was 9.3% (10/108) and 4.3% (9/211), respectively (p=0.07). HIV prevalence was found to be significantly higher among male (n=2024) than among female (n=1702) TB patients (4.4% versus 0.9%; p<0.0001). Only 319 of 641 (49.8%) ICTC patients referred to the chest clinic reached there. Our study highlights the strong need to scale up the integration and partnership between HIV and TB programmes for better and integrated diagnosis and care of HIV-TB co-infected patients.

Hepatitis B, C, and D Virus Infection among Population Aged 10-64 Years in Mongolia: Baseline Survey Data of a Nationwide Cancer Cohort Study.

Hepatitis B, C, and D virus infections are a major public health problem, and Mongolia has one of the highest prevalences of dual and triple infections in the world. We aimed to determine the seroprevalence of hepatitis infection and dual or triple hepatitis infections among 10-64-year-olds. A questionnaire was used to identify risk factors for hepatitis infection, and seromarkers were measured by the fully automated immunologic analyzer HISCL-5000. Among a total of 10,040 participants, 8.1% of the population aged 10-64 was infected with HBV, 9.4% with HCV, and 0.4% with HBV and HCV, and the prevalence of the disease varied by age, sex, and the area of residence. Young people were particularly unaware of their hepatitis infection status. A small proportion of children aged 10 to 19 years and the majority of adults younger than 30 years were unaware of their HBV and HCV infection. Men were also more likely to be unaware of their HBV and HCV infection status than women. The results suggested that the prevalence of infection in the general population is high and that most people are unaware that they are infected or have become chronic carriers. Identifying mono-, co-, or triple-infection status is critical to prevent the rapid progression of liver disease among the Mongolian population.

Immunological Interactions between Intestinal Helminth Infections and Tuberculosis.

Helminth infections are among the neglected tropical diseases affecting billions of people globally, predominantly in developing countries. Helminths' effects are augmented by coincident tuberculosis disease, which infects a third of the world's population. The role of helminth infections on the pathogenesis and pathology of active tuberculosis (T.B.) remains controversial. Parasite-induced suppression of the efficacy of Bacille Calmette-Guerin (BCG) has been widely reported in helminth-endemic areas worldwide. T.B. immune response is predominantly proinflammatory T-helper type 1 (Th1)-dependent. On the other hand, helminth infections induce an opposing anti-inflammatory Th2 and Th3 immune-regulatory response. This review summarizes the literature focusing on host immune response profiles during single-helminth, T.B. and dual infections. It also aims to necessitate investigations into the complexity of immunity in helminth/T.B. coinfected patients since the research data are limited and contradictory. Helminths overlap geographically with T.B., particularly in Sub-Saharan Africa. Each disease elicits a response which may skew the immune responses. However, these effects are helminth species-dependent, where some parasites have no impact on the immune responses to concurrent T.B. The implications for the complex immunological interactions that occur during coinfection are highlighted to inform government treatment policies and encourage the development of high-efficacy T.B. vaccines in areas where helminths are prevalent.

Dactylogyrus extensus and Pseudomonas fluorescens dual infection in farmed common carp (Cyprinuscarpio)

Dactylogyrus extensus and Pseudomonas fluorescens are serious pathogens in Cyprinus carpio aquaculture causing severe impacts and substantial economic losses. During the early spring of 2021, abnormal mortalities were reported among farmed C. carpio. Moribund fish showed anorexia, respiratory distress, dermal ulcers, and septicemia. The water analysis revealed low dissolved oxygen (3.4 mg/L), and high un-ionized ammonia levels (0.65 mg/L). Seventy moribund C. carpio specimens were collected and subjected to parasitological and bacteriological examinations. The monogenetic trematode D. extensus was discovered in wet mounts from the gills of all the examined fish samples (100%). The identity of recovered parasites was confirmed by sequencing and alignment of the 28S rDNA gene. P. fluorescens was concurrently identified in the infested fish samples (58.5%) based on phenotypic characteristics using the API20 E. The identity of bacterial isolates was confirmed further by sequencing and alignment of 16S rRNA gene. The IL-1β and MHCII were upregulated in infested fish in tandem with the severity of infections. P. fluorescens isolates displayed high resistance to most of the tested antibiotics. The study is one of the earlier reports on D. extensus and P. fluorescens co-infections in farmed C. carpio and highlights the need of effective control programs to protect fish health and minimize losses.

Evidence of a novel cross-species transmission by ovine papillomaviruses.

Ovine papillomavirus (OaPV) comprises four genotypes; OaPV1, OaPV2 and OaPV4 are fibropapillomaviruses within the genus Deltapapillomavirus, whereas OaPV3 is an epitheliotropic virus that belongs to the genus Dyokappapapillomavirus. To date, all of them have been known to infect sheep only. OaPV1, OaPV2 and OaPV4 have been associated with ovine cutaneous and mucosal fibropapillomas, whereas OaPV3 is a key factor in the squamous cell carcinoma pathway of the sheep skin. Whole blood samples obtained from 128 cattle at public slaughterhouses were investigated using droplet digital polymerase chain reaction (ddPCR). ddPCR is a new-generation PCR technique that enables an accurate and absolute quantification of target molecules with high sensitivity and specificity. All OaPVs were detected by identification and quantification of nucleic acids using specific fluorescent probes. Of 128 blood samples, 100 (∼78%) showed OaPV infections. Further, 42, 35 and 23 blood samples showed single, double and triple OaPV infections, respectively. OaPV1 was responsible for 22 single infections, OaPV2 caused 16 single infections and OaPV3 and OaPV4 caused two single infections each. OaPV1 and OaPV2 were the most frequent ovine viruses in dual and triple infections. In many blood samples, both ovine deltapapillomavirus and dyokappapapillomaviruswere found to be transcriptionally active, as shown by the detection and quantification of E5 oncogene transcripts for OaPV1, L1 transcripts for OaPV2, E6 and E7 transcripts for OaPV3 and E6 for OaPV4. OaPVs were found in the blood samples from cattle that shared grasslands rich in bracken ferns known to contain immunosuppressant substances. Furthermore, OaPVs were also found in cattle from intensive livestock farming without any contact with sheep. Because OaPV DNA was detected in both grass hay and corn silage, it is conceivable that these feed may be the viral sources.

Virulence Comparison of Four Porcine Circovirus Type 2 (PCV2) Genotypes (2a, 2b, 2d and 2e) in Pigs Single-Infected with PCV2 and Pigs Dual-Infected with PCV2 and Mycoplasma hyopneumoniae

The objective of this study was to compare the virulence of four porcine circovirus type 2 (PCV2) genotypes (2a, 2b, 2d and 2e). Pigs were infected with one of these four genotypes. Pigs were also dually infected with Mycoplasma hyopneumoniae and one of the four PCV2 genotypes. Virulence was determined based on the amount of PCV2 loads in the blood and lymph nodes and the severity of lymphoid lesions. Marked differences in virulence were found among the four genotypes. Within the single infection model, PCV2a, PCV2b and PCV2d were more virulent than PCV2e, while significant differences in virulence were not found among the PCV2a, PCV2b and PCV2d groups. Within the dual infection model, PCV2d was more virulent than the other three PCV2 genotypes. Mycoplasma hyopneumoniae potentiated the severity of PCV2-associated lymphoid lesions and increased the amount of PCV2 loads in the blood and lymph nodes, regardless of the PCV2 genotype. By contrast, PCV2 was not able to potentiate the severity of mycoplasmal-induced lung lesions or the level of M. hyopneumoniae laryngeal load. The results of this study demonstrate that PCV2d is of major clinical importance, while PCV2e is of minor clinical importance.

Molecular typing of dengue virus in Mizoram, Northeast India.

Dengue is an emerging vector-borne public health threat and characterization at the molecular level is important for proper management of the disease. The aim of the study is to examine the diversity of the dengue viral serotypes from a hilly mountainous region of Northeast India. Thirty-six blood samples that were positive for dengue virus IgM antibodies identified by the enzyme-linked immunosorbent assay (ELISA) method were collected and quantified for the IL6 gene expression by using reverse transcriptase polymerase chain reaction (RT-PCR). All the patients had dengue hemorrhagic fever (DHF); 12 samples had a monotypic infection and 14 samples had dual infection with various dengue virus (DENV) serotypes; one sample had triple infection with DENV-1, DENV-2, and DENV-3. This study identified DENV-1 as the major serotype in the state of Mizoram and it is the first report on the molecular typing of the dengue virus from the hilly mountainous state located in the Indo-Burma region bordering Myanmar and Bangladesh.

Effect of Human Immunodeficiency Virus Infection on Human Papillomavirus Clearance Among Women in Senegal, West Africa.

Persistent infection with high-risk human papillomavirus (HPV) is associated with development of invasive cervical cancer. Longitudinal data was collected from 174 Senegalese women. We employed marginal Cox proportional hazards models to examine the effect of human immunodeficiency virus (HIV) status (HIV positive vs HIV negative) and HIV type (HIV-1 vs HIV-2 vs dual HIV-1/HIV-2) on clearance of type-specific HPV infection. Analyses were stratified by incident versus prevalent HPV infection. Incident HPV infections in HIV-positive women were less likely to clear than those in HIV-negative women (adjusted hazard ratio [HR] = 0.60; 95% confidence interval [CI], .38-.94). Among HIV-positive women, HIV-2-infected women and HIV-1/2 dually infected women were more likely to clear HPV incident infections than HIV-1-infected women (HR = 1.66; 95% CI, .95-2.92 and HR = 2.17; 95% CI, 1.12-4.22, respectively). Incident HPV infections in HIV-positive women with CD4 cell count ≤500 cells/μL were less likely to clear than those in HIV-positive women with CD4 cell count >500 cells/μL (HR = 0.65; 95% CI, .42-1.01). No significant associations were observed for prevalent HPV infections. HIV infection reduced the likelihood of clearance of incident HPV infection. Furthermore, among HIV-positive women, low CD4 cell count and dual HIV infection were each associated with reduced likelihood of clearance.

Dual positivity of HIV and anti-HCV in the highly infected population of Rivers State, Nigeria

The study looked to establish dual positivity of human immunodeficiency virus (HIV) and anti-hepatitis C virus (HCV) antibody among HIV-infected individuals in Port Harcourt, Nigeria. Plasma samples from 89 HIV-infected individuals presenting at the Retroviral Clinic of the University of Port Harcourt Teaching Hospital (UPTH), Rivers State, Nigeria, were assayed for anti-HCV-antibody. Seropositivity of anti-HCV-antibody was detected with ELISA kits. Variables tested include sex, age group, educational status, marital status and occupation. Significant variance (p&lt;0.05) existed between patients with mono-HIV infection and those with dual infection of HIV and HCV. Of these infections, 20(22.5%) had HIV and HCV dual infection, while 69(77.5%) had HIV mono-infection. Higher seroprevalence of HIV was found in females [65(73.0%)] than males [24(27.0%)]. The highest seroprevalence of HIV was found in the age group 41-60 years [41(46.1%)], and the age-group 20-30 years had the least prevalence [19(21.3%)]. Also, females had higher HIV and HCV dual positivity (23.1%) than males (20.8%). The age group 31-40 years in this study had the highest HIV and HCV dual positivity (34.5%), while the age group 41-60 years had the lowest prevalence (12.2%). Regarding marital status, singles constituted most study participants and had a dual positivity rate of 28.6% for HIV and HCV. This dual positivity was higher than those divorced and married, with 20.0% and 14.3% prevalence, respectively. Patients with tertiary education (27.3%) and those employed (41.2%) had a higher prevalence than others. None of these demographic characteristics was significantly associated with HIV-HCV dual positivity (p&gt;0.05) except for occupations (p=0.04). Our study further confirms the dual positivity of HIV and anti-HCV in Rivers State, Nigeria. Planned prevention, screening, and treatment are required to reduce further transmission and morbidity.

Prevalence of oral Epstein-Barr virus and Porphyromonas gingivalis and their association with periodontal inflamed surface area: A cross-sectional study.

We previously reported that oral herpesviruses, such as Epstein-Barr virus (EBV), are associated with periodontitis. However, the relationship between oral EBV or dual oral EBV and Porphyromonas gingivalis infections and periodontal inflammation severity remains unclear. We conducted this study to determine the relationship between oral EBV and P gingivalis prevalence and the periodontal inflamed surface area (PISA) in middle-aged and older adults. We analyzed 205 patients (median age, 70 years) who visited Hiroshima University Hospital. Tongue swab samples were used to investigate the presence of EBV and P gingivalis DNA using real-time PCR. Probing pocket depth and bleeding on probing were measured at 6 sites per tooth. PISA scores were calculated based on the results of probing pocket depth and bleeding on probing. Propensity scores were calculated via logistic regression analysis of 8 clinical factors: age, sex, smoking status, remaining teeth, denture use, hypertension, diabetes, and hyperlipidemia. EBV DNA was present in 41 of the 205 participants (20.0%). Thirty-seven EBV-positive or -negative participants in 74 matched pairs after propensity-score matching were examined via univariate analysis. EBV-positive participants exhibited higher plaque control record scores and PISAs than did EBV-negative participants. EBV DNA was significantly associated with plaque control record scores and PISA (both P = .04). Of the 205 participants, 111 were positive for P gingivalis (54.1%). Nineteen participants (9.3%) were infected with both oral EBV and P gingivalis. Logistic regression analysis revealed that dual infection with EBV and P gingivalis was significantly associated with diabetes (odds ratio = 3.37, 95% confidence interval: 1.13-10.1; P = .03). Oral EBV prevalence is associated with oral hygiene and the spread of inflamed periodontal tissue. Diabetes may be a risk factor for dual infection with oral EBV and P gingivalis.

Impact of advanced practice providers on characteristics and quality of care of patients with chronic hepatitis B.

Advanced practice providers (APP) may be able to play a role in improving the linkage to care in patients with chronic hepatitis B (CHB), but data are limited. To compare management of patients with CHB under APP-assisted versus physician-only care METHODS: This retrospective analysis identified patients with CHB infection from Optum's de-identified Clinformatics® Data Mart Database (2003-2021) using ICD-9/ICD-10 codes. We compared the proportion of patients with CHB who had adequate evaluation for treatment (defined as ALT, HBV DNA, ± HBeAg), and the proportion of treatment-eligible patients with CHB who received treatment between APP versus physician-only care. We included 42,140 eligible patients (mean age: 51.9 ± 15.1; 56.1% male). Overall, 34.3% received APP care with increasing utilisation over time. Compared to physician-only care, patients who also received APP care were more likely to have viral co-infection, and more likely to have been seen by a specialist (72.1%). Overall, 62.8% and 56.2% of treatment-eligible patients based on AASLD and EASL guidelines, respectively, received treatment. APP care patients were more likely to be treated (AASLD adjusted HR: 1.18, 95%CI: 1.03-1.34; EASL adjusted HR:1.24, 95%CI: 1.09-1.41) after adjustment for age, sex, race/ethnicity, viral dual infection, baseline cirrhosis/liver cancer, number of HBV DNA and alanine aminotransferase measurements, and physician provider type. Treatment-eligible patients with CHB receiving APP care were more likely to receive antiviral therapy. APP care may help to expand the pool of providers for patients with CHB, and to improve current suboptimal treatment rates.

Dual Fungal Infection of Aspergillosis and Mucormycosis in a COVID-19 Patient: A Rare Case Report

Coronavirus disease 2019 (COVID-19) infections can be related to vast spectrum of co-existent bacterial and fungal infections. A 49-year-old diabetic male was admitted with a history of fever, cough and breathlessness since 5 days. He developed persistent headache with right sided purulent nasal discharge. Relevant histo-pathological, biochemical, microbiological and imaging studies were performed which proved it to be a dual infection of Aspergillosis and Mucormycosis. We present one such case in a COVID-19 patient to highlight its unusual clinical features along with the diagnostic and therapeutic challenges.

Multilocus sequence typing of Giardia duodenalis genotypes circulating in humans in a major metropolitan area.

Giardia duodenalis is an intestinal protozoan parasite of humans and animal hosts and comprises eight microscopically indistinguishable molecularly-diverse lineages designated as assemblages A–H. Assemblages A and B are the primary sources of infections in humans and a wide range of mammals. Here, we identified assemblages, and inter-/intra-assemblage genetic diversity of human G. duodenalis isolates based on the multilocus sequence typing of the triosephosphate isomerase (tpi), β -giardin (bg), and glutamate dehydrogenase (gdh) loci. Multilocus sequence analysis of 62 microscopically-positive G. duodenalis fecal samples identified 26 (41.9%), 27 (43.5%), and nine (14.5%) isolates belonging to assemblages A, B, and discordant assemblages, respectively. The tpi locus assemblage-specific primers identified dual infections with A and B assemblages (45.2%). The sequence analysis of multiple alignments and phylogenetic analysis showed low genetic polymorphism in assemblage A isolates, classified as sub-assemblage AII at three loci, subtype A2 at tpi and gdh loci, and subtype A2 or A3 at bg locus. High genetic variations were found in assemblage B isolates with 14, 15, and 23 nucleotide patterns at tpi, bg, and gdh loci, respectively. Further concatenated sequence analysis revealed four multilocus genotypes (MLG) in 24 assemblages A isolates, two previously-identified (AII-1 and AII-5), with one novel multilocus genotype. However, the high genetic variations observed in assemblage B isolates among and within the three genetic loci prevented the definitive designation of specific MLGs for these isolates. Multilocus sequence typing may provide new insight into the genetic diversity of G. duodenalis isolates in Tehran, suggesting that humans are likely a potential source of G. duodenalis infection. Further host-specific experimental transmission studies are warranted to elucidate the modes of transmission within multiple host populations.

Assessment of multiplex PCR for detection of FMDV, BVDV, BTV, and possible coinfection with Pasteurella multocida in cattle

Multiplex polymerase chain reaction (mPCR) assay is a nucleic acid amplification method that is considered reliable and practical means for several pathogen detections in a single reaction, especially when multiple pathogens are suspected. In this study, a novel mPCR assay was validated for the detection of four notifiable diseases in cattle, including foot and mouth disease (FMD), Bovine viral diarrhea (BVD), Bluetongue (BT), and Hemorrhagic Septicemia (HS). The assay was operated in a two-step procedure. The first one was a reverse transcription of viral RNA, then mPCR of viral cDNA and bacterial DNA. The optimized mPCR was applied on blood (26) and vesicular epithelium (10) samples collected from 26 clinically infected animals from three governorates (Qalubia, Sharkia, and Gharbia). mPCR detected at least 10 pg of microbial nucleic acid extracted from the local isolates. The mPCR results showed that 22/26 (84.6%) of clinically infected animals were positively infected by single or dual infection. Mixed infection of FMDV and Pasteurella multocida was recorded in 11 animals (42.3%), while single FMDV infection was recorded in 5 animals (19.2 %). Single BVDV infection was detected in 5 animals (19. 2 %) and dual infection with FMDV in 1 animal (3.8%). Notably, BTV was not detected in any of the clinical samples. The assessed mPCR was a rapid, accurate, and sensitive test for diagnosing single and mixed infections in cattle and could be used to screen the notifiable diseases affecting cattle.