Concurrent Histomonas meleagridis and Hemorrhagic Enteritis Virus Infection in a Turkey Flock with Recurrent History of Blackhead Disease.

Intestinal health is one of the key factors required for the growth and production of turkeys. Histomoniasis (blackhead disease), caused by a protozoan parasite, Histomonas meleagridis, is a reemerging threat to the turkey industry. Increased incidences of histomoniasis have been reported in recent years due to withdrawal of antihistomonas treatments. H. meleagridis affects ceca and causes cecal inflammation and necrosis. H. meleagridis migrates from ceca to the liver and causes liver necrosis, resulting in high mortalities. Ironically, field outbreaks of histomoniasis are not always associated with high mortalities, while low mortalities have also been documented. There are several exacerbating factors associated with high mortality rates in histomoniasis outbreaks, with concurrent infection being one of them. Recurrent histomoniasis outbreaks in a newly constructed barn were documented, and concurrent infection of H. meleagridis and hemorrhagic enteritis virus was confirmed. Currently, neither commercial vaccines nor prophylactic or therapeutic solutions are available to combat histomoniasis. However, there are treatments, vaccines, and solutions to minimize or prevent concurrent infections in turkeys. In addition to implementing biosecurity measures, measures to prevent concurrent infections are critical steps that the turkey industry can follow to reduce mortality rates and minimize the production and economic losses associated with histomoniasis outbreaks.

Infección simultánea por Histomonas meleagridis y el virus de la enteritis hemorrágica en una parvada de pavos con antecedentes recurrentes de enfermedad de la cabeza negra. La salud intestinal es uno de los factores clave necesarios para el crecimiento y producción de los pavos. La histomoniasis (enfermedad de la cabeza negra), causada por un parásito protozoario, Histomonas meleagridis, es una amenaza reemergente para la industria del pavo. En los últimos años se ha informado de un aumento de la incidencia de histomoniasis debido al retiro de los tratamientos con antihistomonas. Histomonas meleagridis afecta los ciegos y causa inflamación y necrosis cecal. Histomonas meleagridis migra desde los ciegos al hígado y causa necrosis hepática, lo que resulta en una alta mortalidad. Irónicamente, los brotes de histomoniasis en el campo no siempre se asocian con una mortalidad elevada, aunque también se han documentado mortalidades bajas. Hay varios factores exacerbantes asociados con altas tasas de mortalidad en los brotes de histomoniasis, siendo la infección concurrente uno de ellos. Se documentaron brotes recurrentes de histomoniasis en un alojamiento avícola recién construido y se confirmó la infección concurrente de H. meleagridis y el virus de la enteritis hemorrágica. Actualmente no se dis-pone de vacunas comerciales ni soluciones profilácticas o terapéuticas para combatir la histomoniasis. Sin embargo, existen tratamientos, vacunas y soluciones para minimizar o prevenir infecciones concurrentes en los pavos. Además de implementar medidas de bioseguridad, las medidas para prevenir infecciones concurrentes son pasos críticos que la industria del pavo puede seguir para reducir las tasas de mortalidad y minimizar las pérdidas económicas y de producción asociadas con los brotes de histomoniasis.

Evaluation of the Association Between Coronary Artery Aneurysms and Concomitant Infection in Patients With Kawasaki Disease.

We analyzed the relationship between recovery from coronary artery aneurysms (CAAs) and concurrent infections in patients with Kawasaki disease (KD). The estimated median time of aneurysm persistence between patients with and without infections was compared using Kaplan-Meier survival analyses. Risk factors associated with persistent CAAs at 2 years were identified using multivariable analyses. Co-infection was confirmed in 20.5% (106/518) of patients diagnosed with KD. No significant differences regarding treatment or coronary artery outcome were identified between patients with and without infections. The estimated median time of aneurysm persistence was higher in the co-infected group (9 vs. 6 months). A maximum Z-score ≥ 4.00 at 1 month had 78% sensitivity and 83% specificity in predicting CAAs without recovery within 1 year of onset, whereas the predictability was higher within 2 years of onset, with a Z-score ≥ 4.88 (sensitivity, 92%; specificity, 91%). Concomitant infections did not affect the response to treatment or coronary artery outcomes in patients with KD.

The epidemiology of concurrent infection in patients with pyogenic spine infection and its association with early mortality: A nationwide cohort study based on 10,695 patients.

BACKGROUND: Generally, a sufficient duration of relevant antibiotics based on an appropriate culture combined with proper surgical treatment guarantees a favorable clinical outcome in patients with pyogenic spine infections. However, a patient's condition often deteriorates as concurrent infections occur in other organs, leading to mortality. Therefore, this study aimed to investigate the epidemiology of concurrent infections in patients with a pyogenic spine infection and estimate the rates and risks of early mortality. METHODS: Patients with a pyogenic spine infection were identified using a national claims database that includes the entire population. The epidemiology of the six types of concurrent infections was investigated, and the corresponding early mortality rates and risks were estimated. The results were validated internally by bootstrapping and externally by defining two additional cohorts for sensitivity analysis. RESULTS: Among 10,695 patients with a pyogenic spine infection, the prevalence of the six types of concurrent infections was 11.3 % for urinary tract infections, 9.4 % for intra-abdominal infections, 8.5 % for pneumonia, 4.6 % for septic arthritis or osteomyelitis of the extremities, 0.7 % for central nervous system infections, and 0.5 % for cardiac infections. Patients with a concurrent infection had approximately 4-fold greater mortality than those without (3.3 % vs. 0.8 %). The early mortality rates were particularly higher in patients with multiple or specific types of concurrent infections, including central nervous system infections, cardiac infections, and pneumonia. In addition, the mortality trends differed significantly according to the number and type of concurrent infections. CONCLUSIONS: These data on six types of concurrent infection among patients with pyogenic spinal infection can be used as a source of reference by clinicians.

COVID-19 and Early Post-Primary TB: Commonalities of Pathobiology in Pneumonitis and Therapies.

OBJECTIVE: Concurrent infection with COVID-19 and M. tuberculosis has been reported to be more severe than either alone, resulting in increased mortality. Our objective was to define the shared pathobiology of COVID-19 and the developmental stage of TB in the lung and explore adjunctive therapies to treat such commonalities. METHODS: Since morphoproteomics combines the disciplines of histopathology, molecular biology and protein chemistry to paint a portrait of the protein circuitry in diseased cells for the purpose of uncovering targets amenable to specific intervention [1], we used morphoproteomic analyses to study lung tissues of patients with early post-primary tuberculosis or COVID-19 infection. RESULTS: These studies showed co-localization of the COVID-19 virus and M. tuberculosis antigens with cyclo-oxygenase-2 and fatty acid synthase in the reactive alveolar pneumocytes and with programmed death-ligand 1 expression on the alveolar interstitium and alveolar pneumocytes. This was associated with accumulation of pro-infectious M2 polarized macrophages in the alveolar spaces. CONCLUSION: The commonalities in these pathways suggest that they might be susceptible to adjunctive therapies with metformin and vitamin D3. This is supported by published studies that metformin and vitamin D3 could reduce the severity of both COVID-19 and early post-primary TB infections.

Concurrent infections of Streptococcus iniae and Aeromonas veronii in farmed Giant snakehead (Channa micropeltes).

The giant snakehead, Channa micropeltes, is an increasingly important economic freshwater fish in Thailand and other regions of Asia. Presently, giant snakehead are cultured under intensive aquaculture conditions, leading to high stress and conditions favouring disease. In this study, we reported a disease outbreak in farmed giant snakehead with a cumulative mortality of 52.5%, continuing for 2 months. The affected fish exhibited signs of lethargy, anorexia and haemorrhage of the skin and eyes. Further bacterial isolations revealed two different types of colonies on tryptic soy agar: small white, punctate colonies of gram-positive cocci and cream-coloured, round and convex colonies of rod-shaped gram-negative bacteria. Additional biochemical and species-specific PCR analysis based on 16S rRNA confirmed the isolates as Streptococcus iniae and Aeromonas veronii. Multilocus sequence analysis (MLSA) placed the S. iniae isolate into a large clade of strains from clinically infected fish worldwide. Gross necropsy findings showed liver congestion, pericarditis and white nodules in the kidney and liver. Histologically, the affected fish showed focal to multifocal granulomas with inflammatory cell infiltration in kidney and liver, enlarged blood vessels with mild congestion within the meninges of the brain and severe necrotizing and suppurative pericarditis with myocardial infarction. Antibiotic susceptibility tests revealed that S. iniae was sensitive to amoxicillin, erythromycin, enrofloxacin, oxytetracycline, doxycycline and resistant to sulfamethoxazole-trimethoprim, while the A. veronii was susceptible to erythromycin, enrofloxacin, oxytetracycline, doxycycline, sulfamethoxazole-trimethoprim and resistant to amoxicillin. Conclusively, our findings highlighted the natural concurrent bacterial infections in cultured giant snakehead, which support the implementation of appropriate treatment and control strategies.

Occurrence of Dengue Virus Infection with Multiple Serotypes in Central Karnataka, India.

Objective To find the incidence of concurrent infection of dengue and correlate the difference in clinical features, laboratory diagnoses, and outcomes between dengue and dengue-like illnesses. Methodology A total of 2,256 patients with suspected dengue fever during the period of July 2014 to June 2020 as per the WHO case definition for dengue fever were enrolled in the study. All patients admitted with a suspected dengue fever were studied in detail in terms of demographic features, clinical features, and laboratory profiles. Two blood samples were collected from each patient with a history of fever for 5 to 7 days. Investigation consisted of CBC, Widal test, malarial card test, Weil Felix test, Scrub typhus test, chikungunya, dengue parameters such as dengue NS1, IgM, and IgG, and real-time PCR for dengue serotypes were performed for each sample. Results A total of 1,412 males (62.6%) and 844 females (37.4%) of age 2 to 44 years were hospitalized. Out of 2,256 clinically suspected dengue cases, 1,306 cases were positive and 950 were negative by RT-PCR test. Fever was the most common clinical features among the RT-PCR-positive cases, followed by retroorbital pain (85.9%), flushing in 77.5%, and rashes in 84.8% of patients. ARDS was seen in 9.7% and splenomegaly in 27.5% patients. A platelet count of less than 100,000 was observed in 1,838 (81.5%) patients, and a platelet count of less than 20,000 was observed in 147 (6.5%) patients. Of 2,256 samples, 1,306 (57.9%) tested positive for dengue viral RNA by RT-PCR. Also, 798 cases were infected with a single DENV serotype, and 608 had a concurrent infection. Of the 798 single DENV serotype infection cases, 392 (54.2%) were typed as DENV-2 and 218 (29.2%) as DENV-3. Coinfection with serotypes DENV-2 and DENV-3 was found to account for 67.8% of all concurrent infections. Conclusion The study showed that dengue fever with concurrent infection with multiple serotypes is on the rise, and an occurrence of recombination may lead to the emergence of more virulent strains showing varied clinical presentations.

Molecular evidence for concurrent infection of goats by orf virus and bovine herpesvirus 1.

Orf is a disease of small ruminant animals, including goats and sheep, that is caused by a parapoxvirus. Although the mortality rate is low, economic losses may occur due to the clinical signs. Bovine herpesvirus 1 (BoHV-1) infection is known to cause respiratory and reproductive disorders mainly in cattle; however, it has been found to circulate among goats and sheep as well. In contrast to orf virus (ORFV), BoHV-1 does not induce clinical disease in goats. In this study, we aimed to detect the presence of ORFV by molecular methods and to uncover eventual simultaneous herpesvirus infections masked by orf disease signs. To this end, 82 goats, housed near to a cattle herd, were tested. By polymerase chain reaction (PCR), three goats (3.7%) were found to harbour both viruses, while an additional goat was positive for ORFV only. The PCR products were sequenced and phylogenetic analyses were performed. This study revealed that ORFV and BoHV-1 may be present simultaneously in an animal causing a concurrent infection. These data should be taken into consideration when looking for secondary pathogens in diseased goats, and the prevention methods should be developed accordingly.

Clinical Characteristics of Patients with Pyogenic Vertebral Osteomyelitis and Concurrent Infections and Their Clinical Outcomes.

Patients with pyogenic vertebral osteomyelitis (PVO) often develop concurrent infections, and a significant number of these patients show rapid deterioration in their medical condition, leading to mortality without PVO-related structural instability or neurological deficits. To improve clinical outcomes, we investigated the clinical presentation and treatment outcomes of patients with PVO and concurrent infections. This study included 695 patients with PVO, of which 175 (25%) had concurrent infections and 520 (75%) did not. The clinical characteristics of the two groups were compared, and multivariable analysis was performed to identify the association between concurrent infections and clinical outcomes. Patients with concurrent infections were older and had more comorbidities than those without. Moreover, there were significant intergroup differences in the anatomical involvement of PVO, and patients with concurrent infections had a higher number of regions involved more frequently than those without concurrent infections (15% vs. 6%). In contrast, patients with concurrent infections showed a lower degree of focal invasiveness, including a lower incidence of posterior abscess (47% vs. 59%; p = 0.008) and fewer neurological impairments according to the American Spinal Injury Association grade (p < 0.001) than those without concurrent infections. The causative organisms also differed significantly between the two groups, and patients with concurrent infections had a greater proportion of Gram-negative infections (31% vs. 16%, respectively) and a smaller proportion of methicillin-resistant S. aureus infections than those without concurrent infections (6% vs. 24%). Consequently, their clinical outcomes were significantly different, and patients with concurrent infections showed lower recurrence and higher mortality rates. We investigated the 1-year recurrence and mortality rates and their 95% confidence intervals according to the types of concurrent infections and their time of diagnosis and found variations in these parameters. Our results, based on a large number of patients, can be practically used as a reasonable reference to warn clinicians of the clinical risks of concurrent infections in patients with PVO and to help predict their clinical outcomes.

Eimeria pragensis infection alters the gut microenvironment to favor extrinsic shiga toxin-producing Escherichia coli O157:H7 colonization in mice.

We examined the effects of Eimeria pragensis infection on intestinal peristalsis, goblet cell proliferation and intestinal flora in C57BL/6 mice. Intestinal peristalsis was evaluated by radiography using barium at 7 days post-infection (p.i.). The intestinal peristalsis of E. pragensis-infected mice was significantly suppressed compared with uninfected control mice. Twenty-three mice were divided into 5 groups of 4 or 5 mice each; 2 groups of mice were infected with E. pragensis and the others were kept uninfected. At 7 days p.i., E. pragensis-infected and -uninfected mice were sacrificed to examine goblet cell numbers in the intestines, and significant decreases were observed only in the infected mice. Shiga toxin-producing Escherichia coli (STEC) O157:H7 was inoculated orally in mice both infected and uninfected with E. pragensis at 7 days p.i., with the remaining mice used as uninoculated controls. When mice were sacrificed at 2 days after STEC inoculation, STEC was only detected in the intestines of E. pragensis-infected mice. Colonization of STEC was also confirmed by immunohistochemistry on the surface of epithelial cells in concurrently infected/inoculated mice. Also, an overgrowth of residential E. coli was observed only in E. pragensis-infected mice. These results suggest that E. pragensis induces the suppression of intestinal peristalsis and modifies the intestinal environment to facilitate artificially introduced STEC colonization and multiplication, in addition to residential E. coli overgrowth.

Cryptosporidium spp. Infections in Combination with Other Enteric Pathogens in the Global Calf Population.

The most common worldwide diarrhoea-causing agents in neonatal calves are Cryptosporidium spp. (Crypto), bovine rotavirus (BRV), bovine coronavirus (BCoV), and enterotoxigenic Escherichia coli F5 (K99) (ETEC). Crypto is a zoonotic pathogen of diarrhoea in humans, particularly for children and immunocompromised adults. Four weighted-stratified random-effect meta-analyses including meta-regression analyses were performed to calculate the worldwide mean prevalence of Crypto and associated concurrent infections with BRV, BCoV and ETEC, as well as their potential influencing factors. The meta-analysis incorporated 28 studies (56 substudies) in 17 countries that determined the presence or absence of concurrent infections with Crypto in the global calf population. Approximately half of all considered studies presented here were conducted in Europe independently of the type of infections with Crypto. Within Europe, the highest estimated mean Crypto-BRV prevalence was identified in Ireland (16.7%), the highest estimated mean Crypto-BCoV prevalence was detected in the United Kingdom (4.3%), and the highest estimated mean Crypto-ETEC prevalence across the literature was determined in Turkey (4.7%). The chance of detecting BRV, BCoV, and ETEC in calves with diarrhoea was 0.8 (confidence interval (CI): 0.6-1.0), 0.7 (CI: 0.5-1.0) and 0.6 (CI: 0.4-0.9) lower in the presence of Crypto compared to calves without Crypto. This may indicate an inhibitory effect between BRV, BCoV, ETEC, and Crypto in calves. The variance in the published prevalence across the literature can mainly be explained by the "diagnostic" factor (R2 min-max: 0.0-40.3%), followed by the "health status of the sampled animals" (R2 min-max: 1.4-27.3%) and "geographical region" (R2 min-max: 5.9-23.6%).

Streptococcus, Centrocestus formosanus and Myxobolus tilapiae concurrent infections in farmed Nile tilapia (Oreochromis niloticus).

Stress triggered concurrent microbial/parasitic infections are prevalent in earthen pond based farmed Nile tilapia Oreochromis niloticus. In the current study, a total of thirty five O. niloticus were collected from a commercial fish farm with a history of severe mortalities at Port Said, Egypt. Nile tilapia samples were subjected to bacteriological, parasitological and pathological examinations. Twenty one Enterococcus fecalis and 15 Streptococcus agalactiae isolates were presumptively identified utilizing the semi-automated API 20 Strept test kit. The identities of the retrieved bacteria were confirmed by the sequencing of 16 S rRNA gene. Moribund O. niloticus were found to be heavily infected by one or both of Centrocestus formosanus encysted metacercariae (EMC) and/or Myxobolus tilapiae spores presenting a unique form of synergistic and/or symbiotic relationship. The identities of both parasites were confirmed through morphological and molecular characterization. Variable circulatory, degenerative, necrotic and proliferative changes were also noticed in hematopoietic organs. Interestingly, multiple myxobolus spores and EMC were noticed in some histological sections. It was obvious that the current concurrent bacterial and parasitic infections are triggered by the deleterious effects of some stressing environmental conditions. The unfavorable climatic conditions (high temperature and high relative humidity) recorded at the surge of mortalities are probable predisposing stress factors.

Prevalence of Worldwide Neonatal Calf Diarrhoea Caused by Bovine Rotavirus in Combination with Bovine Coronavirus, Escherichia coli K99 and Cryptosporidium spp.: A Meta-Analysis.

Multiple enteropathogens such as bovine rotavirus (BRV), bovine coronavirus (BCoV), Escherichia coli K99 (ETEC) and Cryptosporidium spp. (Crypto) are the most common causes of calf diarrhoea during the first 30 days of animal age. Three weighted-stratified random-effects meta-analyses were performed to calculate the worldwide prevalence of mixed infections of the causative agents (i.e., BRV-BCoV, BRV-ETEC, BRV-Crypto) and their potential influencing factors. The meta-analysis covered 41 studies (94 sub-studies) in 21 countries that determined the presence or absence of mixed infections in global calf populations. The highest worldwide estimated pooled prevalence was identified for BRV-Crypto (6.69%), followed by BRV-BCoV (2.84%), and BRV-ETEC (1.64%). The chance of detecting BCoV in calves with diarrhoea was 1.83 higher in the presence of BRV compared to calves without BRV, whereby an inhibition effect (odds ratio: 0.77) was determined between BRV and Crypto infections. The diagnostic methods were identified as a significant influencing factor in the detection of all considered mixed infections, while the other analysed factors differed in relation to their effect on prevalence. In contrast to BRV-BCoV, the prevalence of BRV-ETEC and BRV-Crypto mixed infections followed the course of individual ETEC and Crypto prevalence related to the age class of the sampled animals.

A Longitudinal Case Study of Concurrent Infection with Syphilis and Human Immunodeficiency Virus During the Early Phase.

Due to the low incidence of concurrent human immunodeficiency virus (HIV) and syphilis infection identified during the early phase, such as window period (WP), little is known about the clinical manifestations, diagnosis, and treatment efficacy at very early stages. One longitudinal study was conducted in a 42-year-old blood donor who was concurrently infected with syphilis and HIV. This blood donor was treated with a penicillin-based regimen and early antiretroviral therapy (ART). Sequential serological and nucleic acid tests were performed and the results were comparatively analyzed. A regular male donor who had two occasions of high-risk sexual behaviors 41 and 35 days before donation donated whole blood at the Shenzhen Blood Center. ART was initiated at the 28th day after donation (DAD), and syphilis treatment was received at the 49th DAD. Microbiological analysis using a fourth-generation anti-HIV enzyme-linked immunosorbent assay (ELISA) (4th GAHE) and electro-chemiluminesent immunoassay indicated a positive signal at the 6th DAD, while a third-generation anti-HIV ELISA (3rd GAHE) showed positive at the 26th DAD. All nucleic acid testing (NAT) for HIV RNA were reactive except the minipool NAT of 6 pooled samples at 117th DAD. The HIV viral load declined more than 4-log in copies per milliliter over 3 months, until reaching nondetectable levels at 246th DAD. Nevertheless, HIV-1 DNA was still detectable at 403rd DAD. Among all methods utilized, anti-treponema pallidum ELISA detected syphilis infection at the earliest time. A successful serological response to syphilis treatment was reached around the 80th DAD. Concurrent infection with syphilis and HIV during early phases did not significantly change the sensitivity of reagents in detection nor alter the therapeutic efficacy for the treatment of both pathogens, but might result in delayed HIV serological WP.

Concurrent Infection of Fascioliasis andTrypanosomosis and Associated Risk Factors in Local Zebu Breed Cattle of Western Ethiopia.

BACKGROUND: A cross-sectional study was conducted from late October 2016 to June 2017, with the primary objective of estimating and analyzing the concurrent occurrence of both fascioliasis and trypanosomosis infections and associated risk factors along the tsetse-infested Didessa river basin. METHODS: The methodology applied was based on stratified sampling for the parasitological study, with entomological and malacological surveys, including fly dissection. RESULTS: The result of variance-ratio testing between trypanosomosis and fascioliasis infections (mean prevalence 0.117±0.322 and 0.283±0.451, respectively), was statistically significant (P[F>f]=0), with higher observed fascioliasis infection (n=147, 28.27%). Severe anemia was observed in trypanosomosis infection, with mean packed cell volume of 19.57 (OR=0.71, P>|z|=0.000), and vast fascioliasis infections identified among cattle with medium and poor body condition in terms of weight (n=91 [32.73%] and n=38 [21.47%]). On entomological study, 578 (62.62%) and 345 (37.38%) female and male Glossina tachinoides fly species were cached, respectively, with overall mean flies/trap/day of 5.19 (n=923). Despite the prevalence of trypanosomosis in infected cattle, of 130 G. tachinoides flies dissected, only three were found to be positive for an infection rate of 2.31%. Malacological study identified three snail species known to maintain fascioliasis: Lymnea truncatula (n=28, 45.16%), Lymnea natalensis (n=23, 37.10%), and Biomphalaria (n=11, 17.74%). Concurrent infection with fascioliasis and trypanosomosis was mainly associated with the co-occurrence of their intermediate host snails and Glossina flies, respectively, with 4.42% (n=23) prevalence. CONCLUSION: This study clearly demonstrated that the former parasite was highly associated with emaciation, whereas the second was responsible for anemia. In future,researchers should focus solely on estimating meat and milk production of local cattle to assess the economic impact of the study parasites.

End Point Multiplex PCR for Diagnosis of Haemoprotozoan Diseases in Cattle.

BACKGROUND: Theileriosis, trypanosomosis and babesiosis are the three major haemoprotozoan diseases causing huge economic losses worldwide. Difficulty in diagnosis of these diseases lies with the detection of carrier state with low parasitemia and concurrent infection. PURPOSE: The present study was conducted to standardize and evaluate multiplex PCR assay for specific, fast and simultaneous detection of Theileria annulata, Trypanosoma evansi and Babesia bovis in bovines. METHODS: Positive parasitic DNA was obtained from microscopically positive samples. Simplex PCR assay was developed targeting repetitive nucleotide sequences for Trypanosoma evansi and gene coding enzyme carbamoyl phosphate synthetase II for Babesia bovis. For theileriosis conditions already standardized targeting cytochrome b gene was used. Gradient PCR assay was used to determine common amplification conditions and develop multiplex PCR assay. Limit of detection was determined using tenfold serial dilution of parasitic DNA. Blood samples collected from 117 bovines suspected for haemoparasite infection was tested by simplex and multiplex PCR assay. RESULTS: Simplex PCR assay was able to detect Theileria annulata, Trypanosoma evansi and Babesia bovis at dilution 10-9, 10-8 and 10-8 which corresponds to copy number 1, 10 and 10, respectively, whereas of multiplex PCR assay was found to be 10-7 dilution corresponding to 100 copy number. PCR products bands obtained in multiplex PCR assay at 257, 312 and 446 bp were easily distinguishable. Results of simplex PCR assay for detection of individual parasites revealed 48 (41.02%), 27 (23.07%) and 5 (4.27%) samples positive for T. annulata, T. evansi and B. bovis, respectively. Sixty-three (53.8%) samples were found positive by multiplex PCR assay with 15 samples (23.8%) showing mixed infection. CONCLUSION: Multiplex PCR assay was found to be highly specific and can be used for easy, early, sensitive, specific and simultaneous diagnosis of haemoprotozoan diseases in epidemiological survey as a robust tool.

Metatranscriptomic Characterization of Coronavirus Disease 2019 Identified a Host Transcriptional Classifier Associated With Immune Signaling.

BACKGROUND: The recent identification of a novel coronavirus, also known as severe acute respiratory syndrome coronavirus 2, has caused a global outbreak of respiratory illnesses. The rapidly developing pandemic has posed great challenges to diagnosis of this novel infection. However, little is known about the metatranscriptomic characteristics of patients with coronavirus disease 2019 (COVID-19). METHODS: We analyzed metatranscriptomics in 187 patients (62 cases with COVID-19 and 125 with non-COVID-19 pneumonia). Transcriptional aspects of 3 core elements, pathogens, the microbiome, and host responses, were evaluated. Based on the host transcriptional signature, we built a host gene classifier and examined its potential for diagnosing COVID-19 and indicating disease severity. RESULTS: The airway microbiome in COVID-19 patients had reduced alpha diversity, with 18 taxa of differential abundance. Potentially pathogenic microbes were also detected in 47% of the COVID-19 cases, 58% of which were respiratory viruses. Host gene analysis revealed a transcriptional signature of 36 differentially expressed genes significantly associated with immune pathways, such as cytokine signaling. The host gene classifier built on such a signature exhibited the potential for diagnosing COVID-19 (area under the curve of 0.75-0.89) and indicating disease severity. CONCLUSIONS: Compared with those with non-COVID-19 pneumonias, COVID-19 patients appeared to have a more disrupted airway microbiome with frequent potential concurrent infections and a special trigger host immune response in certain pathways, such as interferon-gamma signaling. The immune-associated host transcriptional signatures of COVID-19 hold promise as a tool for improving COVID-19 diagnosis and indicating disease severity.

Concurrent infection of Avibacterium paragallinarum and fowl adenovirus in layer chickens.

The diagnosis of a concurrent infection of Avibacterium paragallinarum and fowl adenovirus (FAdV) in an infectious coryza-like outbreak in the outskirt of Beijing is reported. The primary signs of the infection were acute respiratory signs, a drop in egg production, and the presence of hydropericardium-hepatitis syndrome-like gross lesions. Laboratory examination confirmed the presence of A. paragallinarum by bacterial isolation and a species-specific PCR test. In addition, conventional serotyping identified the isolates as Page serovar A. Fowl adenovirus was isolated from chicken liver specimen and identified by hexon gene amplification. In addition, histopathologic analysis and transmission electron microscopy examination further confirmed the presence of the virus. Both hexon gene sequencing and phylogenetic analysis defined the viral isolate as FAdV-4. The pathogenic role of A. paragallinarum and FAdV was evaluated by experimental infection of specific-pathogen-free chickens. The challenge trial showed that combined A. paragallinarum and FAdV infection resulted in more severe clinical signs than that by FAdV infection alone. The concurrent infection caused 50% mortality compared with 40% mortality by FAdV infection alone and zero mortality by A. paragallinarum infection alone. To our knowledge, this is the first report of A. paragallinarum coinfection with FAdV. The case implies that concurrent infections with these 2 agents do occur and more attention should be given to the potential of multiple agents during disease diagnosis and treatment.

Coinfection with Human Norovirus and Escherichia coli O25:H4 Harboring Two Chromosomal blaCTX-M-14 Genes in a Foodborne Norovirus Outbreak in Shizuoka Prefecture, Japan.

ABSTRACT: Hospital-acquired infections caused by extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli are a global problem. Healthy people can carry ESBL-producing E. coli in the intestines; thus, E. coli from healthy people can potentially cause hospital-acquired infections. Therefore, the transmission routes of ESBL-producing E. coli from healthy persons should be determined. A foodborne outbreak of human norovirus (HuNoV) GII occurred at a restaurant in Shizuoka, Japan, in 2018. E. coli O25:H4 was isolated from some of the HuNoV-infected customers. Pulsed-field gel electrophoresis showed that these E. coli O25:H4 strains originated from one clone. Because the only epidemiological link among the customers was eating food from this restaurant, the customers were concurrently infected with E. coli O25:H4 and HuNoV GII via the restaurant food. Whole genome analysis revealed that the E. coli O25:H4 strains possessed genes for regulating intracellular iron and expressing the flagellum and flagella. Extraintestinal pathogenic E. coli often express these genes on the chromosome. Additionally, the E. coli O25:H4 strains had plasmids harboring nine antimicrobial resistance genes. These strains harbored ESBL-encoding blaCTX-M-14 genes on two loci of the chromosome and had higher ESBL activity. Multilocus sequence typing and fimH subtyping revealed that the E. coli O25:H4 strains from the outbreak belonged to the subclonal group, ST131-fimH30R, which has been driving ESBL epidemics in Japan. Because the E. coli O25:H4 strains isolated in the outbreak belonged to a subclonal group spreading in Japan, foods contaminated with ESBL-producing E. coli might contribute to spreading these strains among healthy persons. The isolated E. coli O25:H4 strains produced ESBL and contained plasmids with multiple antimicrobial resistance genes, which may make it difficult to select antimicrobials for treating extraintestinal infections caused by these strains.

Pulmonary tuberculosis and pneumocystis jirovecii concurrent pneumonia in HIV infected patients at a resource limited setting in Eastern India: A case series.

Pneumocytis jirovecii pneumonia (PJP) and Pulmonary TB (PTB) both are common opportunistic infections among HIV infected individuals. But concurrent infections pose a diagnostic challenge owing to similar clinical features. Data suggests a high prevalence of such concurrent infections in developing countries but limited diagnostic modalities especially in resource constraint setup limits accurate diagnosis. At our centre we came across 6 newly diagnosed PTB patients among HIV infected ones had persistent shortness of breath (SOB) and hypoxia despite starting anti-tuberculous treatment (ATT). We excluded concomitant bacterial pneumonia by imaging, sputum examination and blood culture. Serum lactate dehydrogenase (LDH) was estimated and hypoxia by arterial blood gas (ABG). We found all 6 patients had elevated serum LDH, hypoxia and imaging suggestive of PJP were offered sputum for Geisma stain and standard treatment for PJP in form of Bactrim-double strength and steroid. 1 patient had PJ cysts in sputum. 5 patient's classical radiologic findings in form of ground glass opacities in lower lobes along with bilateral infiltrates and 1 had honeycombing. Serum LDH was elevated all 6 subjects. 5 were newly diagnosed HIV and 4 had CD4 count below 50 cells/mm3 and 2 had below 200 cells/mm3.1 patient had developed bilateral pneumothorax as complication. 4 patients responded to treatment and 2 (33.3%) died of respiratory failure during treatment. We were able to diagnose only severe PJP cases as concurrent infection with PTB as there was no availability of broncho alveolar lavage (BAL) as well as direct fluorescent antigen (DFA) test for PJ detection. A high index of suspicion for PJP even in PTB patients with low CD4 count will guide to appropriate therapy for both infections and eventually reduces morbidity and mortality.

Simultaneous detection of scale drop disease virus and Flavobacterium columnare from diseased freshwater-reared barramundi Lates calcarifer.

Freshwater farming of barramundi Lates calcarifer in Thailand is a growing sector in aquaculture, but mortalities due to infectious diseases are still a major threat to this industry. In 2018, an episode of severe mortality in juvenile barramundi was noted in a freshwater earth pond site. Fish presented with severe gill necrosis, as well as severe cutaneous hemorrhages, scale loss, and discoloration at the base of dorsal fin (saddleback lesions). Histopathology revealed extensive necrosis of skeletal muscle and gill filaments, as well as basophilic inclusion bodies and megalocytosis in muscle, gill, liver, and kidney. Scale drop disease virus (SDDV) infection was subsequently confirmed by virus-specific semi-nested PCR. Further, DNA sequences of the viral major capsid protein (MCP) and ATPase genes had a respective homology of 99.85 and 99.92% with sequences of SDDV infecting barramundi in saltwater culture. Gill necrosis and saddleback lesions are not typical lesions associated with scale drop syndrome. Their presence was explained by Flavobacterium columnare isolation from the gill, followed by positive F. columnare-specific PCR. To our knowledge, this is the first report of SDDV-associated mortality in freshwater-farmed barramundi. Furthermore, this mortality presented as a concurrent infection with SDDV and F. columnare, with typical lesions of both infections.