Pathogens

16 pages, 1188 KiB

Open AccessArticle

Genetic Characterization of Palyam Serogroup Viruses Isolated in Japan from 1984 to 2018 and Development of a Real-Time RT-PCR Assay for Broad Detection of Palyam Serogroup Viruses and Specific Detection of Chuzan (Kasba) and D’Aguilar Viruses

by Hiroaki Shirafuji, Natsumi Kishida, Katsunori Murota, Yuto Suda and Tohru Yanase

Pathogens 2024, 13(7), 550; https://doi.org/10.3390/pathogens13070550 (registering DOI) - 28 Jun 2024

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Abstract

We performed whole genome sequencing (WGS) of 15 Palyam serogroup virus (PALV) strains isolated from cattle or Culicoides biting midges in Japan from 1984 to 2018. We found that the PALV strains consisted of Chuzan (Kasba) virus (CHUV), D‘Aguilar virus (DAGV), Bunyip Creek [...] Read more.

We performed whole genome sequencing (WGS) of 15 Palyam serogroup virus (PALV) strains isolated from cattle or Culicoides biting midges in Japan from 1984 to 2018. We found that the PALV strains consisted of Chuzan (Kasba) virus (CHUV), D‘Aguilar virus (DAGV), Bunyip Creek virus, and another PALV, Marrakai virus (MARV). The Japanese MARV strains isolated in 1997 were closely related to Australian PALV strains isolated in 1968–1976 in genome segments 2 and 10, but they were most closely related to other Japanese PALV strains in the other genome segments. Our data suggest that the Japanese MARV strains were reassortant viruses between Asian and Australian PALVs. In addition to the WGS, we developed a real-time reverse-transcription polymerase chain reaction assay that can broadly detect PALV and specifically detect CHUV and DAGV, utilizing the data obtained by the WGS in this study. We detected the DAGV gene in bovine stillborn fetuses and congenitally abnormal calves in 2019 using the newly developed assay. To our knowledge, this is the first report of isolation of MARV outside of Australia and the first report of detection of PALV in bovine fetuses or calves with congenital abnormality outside of Africa. Full article

(This article belongs to the Section Viral Pathogens)

17 pages, 1899 KiB

Open AccessArticle

Sunflower Oil and Cholesterol Nanoemulsion: A Novel Carrier for Micafungin to Combat Multi-Resistant Candida auris

by Gabriel Davi Marena, Alejandro López, Gabriela Corrêa Carvalho, María del Pilar Marín, María Dolores Pérez Ruiz, Jose Manuel Pérez-Royo, María Ángeles Tormo-Mas, Patricia Bernabé, Eulogio Valentín, Taís Maria Bauab, Marlus Chorilli, Javier Pemán and Alba Ruiz-Gaitán

Pathogens 2024, 13(7), 549; https://doi.org/10.3390/pathogens13070549 (registering DOI) - 28 Jun 2024

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Abstract

Candida auris is an emerging, multidrug-resistant yeast that causes systemic infections, mainly in hospitalized or immunosuppressed patients. This pathogen has a high mortality and morbidity rate. This study aims to evaluate the antifungal potential of micafungin (MICA) encapsulated in a nanoemulsion (NEM) against [...] Read more.

Candida auris is an emerging, multidrug-resistant yeast that causes systemic infections, mainly in hospitalized or immunosuppressed patients. This pathogen has a high mortality and morbidity rate. This study aims to evaluate the antifungal potential of micafungin (MICA) encapsulated in a nanoemulsion (NEM) against four clades of C. auris and other non-C. auris species. The antifungal potential of MICA and NEM was evaluated by determining mature biofilm inhibition (0.78–50 µg/mL). The antifungal activities of MICA and NEM (5.92 mg/Kg) were evaluated using an in vivo model of Galleria mellonella. The results showed that NEM intensified the antibiofilm action of MICA, especially in 48 h mature biofilms. In vivo results displayed a higher effectiveness of NEM against all clades of C. auris tested, inhibiting the fungal load in the hemolymph and tissues of G. mellonella with a difference of 3 log10. In addition, C. auris infection caused granulomas surrounded by hemocytes, mainly at the lower and upper ends. Conversely, C. albicans developed pseudohyphae, biofilms, filaments, and chlamydospores. In conclusion, encapsulation of MICA in a nanoemulsion enhances its antifungal activity against mature biofilms of C. auris. This strategy may be considered a therapeutic approach for the control of infections and the dissemination of this new global health threat. Full article

(This article belongs to the Section Fungal Pathogens)

19 pages, 531 KiB

Open AccessReview

Endophytic Trichoderma: Potential and Prospects for Plant Health Management

by Dimitrios Natsiopoulos, Eleni Topalidou, Spyridon Mantzoukas and Panagiotis A. Eliopoulos

Pathogens 2024, 13(7), 548; https://doi.org/10.3390/pathogens13070548 (registering DOI) - 28 Jun 2024

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Abstract

The fungus Trichoderma is widely regarded as the most common fungal biocontrol agent for plant health management. More than 25 Trichoderma species have been extensively studied and have demonstrated significant potential in inhibiting not only phytopathogen growth but also insect pest infestations. In [...] Read more.

The fungus Trichoderma is widely regarded as the most common fungal biocontrol agent for plant health management. More than 25 Trichoderma species have been extensively studied and have demonstrated significant potential in inhibiting not only phytopathogen growth but also insect pest infestations. In addition to their use as biopesticides, there is increasing evidence that several Trichoderma species can function as fungal endophytes by colonizing the tissues of specific plants. This colonization enhances a plant’s growth and improves its tolerance to abiotic and biotic stresses. In recent decades, there has been a proliferation of literature on the role of Trichoderma endophytes in crop protection. Although the mechanisms underlying plant–fungal endophyte interactions are not yet fully understood, several studies have suggested their potential application in agriculture, particularly in the mitigation of plant pests and diseases. This review focuses on the diversity of Trichoderma endophytic strains and their potential use in controlling specific diseases and pests of crop plants. Trichoderma endophytes are considered a potential solution to reduce production costs and environmental impact by decreasing reliance on agrochemicals. Full article

(This article belongs to the Special Issue Fungal Pathogens of Crops)

18 pages, 580 KiB

Open AccessReview

Should Empiric Anti-Fungals Be Administered Routinely for All Patients with Perforated Peptic Ulcers? A Critical Review of the Existing Literature

by Kai Siang Chan, Lee Yee Calista Tan, Sunder Balasubramaniam and Vishal G. Shelat

Pathogens 2024, 13(7), 547; https://doi.org/10.3390/pathogens13070547 (registering DOI) - 28 Jun 2024

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Abstract

A perforated peptic ulcer (PPU) is a surgical emergency with a high mortality rate. PPUs cause secondary peritonitis due to bacterial and fungal peritoneal contamination. Surgery is the main treatment modality and patient’s comorbidites impacts perioperative morbidity and surgical outcomes. Even after surgery, [...] Read more.

A perforated peptic ulcer (PPU) is a surgical emergency with a high mortality rate. PPUs cause secondary peritonitis due to bacterial and fungal peritoneal contamination. Surgery is the main treatment modality and patient’s comorbidites impacts perioperative morbidity and surgical outcomes. Even after surgery, resuscitation efforts should continue. While empiric antibiotics are recommended, the role of empiric anti-fungal treatment is unclear due to a lack of scientific evidence. This literature review demonstrated a paucity of studies evaluating the role of empiric anti-fungals in PPUs, and with conflicting results. Studies were heterogeneous in terms of patient demographics and underlying surgical pathology (PPUs vs. any gastrointestinal perforation), type of anti-fungal agent, timing of administration and duration of use. Other considerations include the need to differentiate between fungal colonization vs. invasive fungal infection. Despite positive fungal isolates from fluid culture, it is important for clinical judgement to identify the right group of patients for anti-fungal administration. Biochemistry investigations including new fungal biomarkers may help to guide management. Multidisciplinary discussions may help in decision making for this conundrum. Moving forward, further research may be conducted to select the right group of patients who may benefit from empiric anti-fungal use. Full article

12 pages, 1728 KiB

Open AccessArticle

The Triterpenoid MOMORDIN-Ic Inhibits HCMV by Preventing the Initiation of Gene Expression in Eukaryotic Cells

by Eleanor Bradley, Emma Poole and Matthew B. Reeves

Pathogens 2024, 13(7), 546; https://doi.org/10.3390/pathogens13070546 - 28 Jun 2024

Viewed by 191

Abstract

Human cytomegalovirus (HCMV) primary infection, re-infection, and reactivation from latency cause morbidity in immune-compromised patients. Consequently, potential therapeutic strategies remain of interest for the treatment of infection. Naturally occurring triterpenoids derived from plants have been demonstrated to have anti-viral activity, although their precise [...] Read more.

Human cytomegalovirus (HCMV) primary infection, re-infection, and reactivation from latency cause morbidity in immune-compromised patients. Consequently, potential therapeutic strategies remain of interest for the treatment of infection. Naturally occurring triterpenoids derived from plants have been demonstrated to have anti-viral activity, although their precise mechanisms of action are not always fully understood. Here, we investigate the activity of Mormordin Ic (Mc) and demonstrate that it is potently anti-viral against HCMV. Through investigation of the mechanistic basis of this anti-viral activity, we identify that it is inhibitory to both viral and host gene expression, and to highly induced genes in particular. We go on to observe that Mc impacts on RNA Pol II activity and, specifically, reduces the occupancy of elongating RNA Pol II at a viral promoter. Next, we demonstrate that Mc is inhibitory to HCMV reactivation, and in doing so identify that it has greater activity against the canonical major immediate early promoter compared to the alternative ip2 promoter located downstream. Finally, we see evidence of RNA Pol II occupancy at the ip2 promoter in undifferentiated myeloid cells. Thus, Mc is potently anti-viral and a potential tool to probe the activity of multiple promoters considered important for controlling HCMV reactivation. Full article

(This article belongs to the Special Issue Understanding Human Cytomegalovirus Pathogenesis: Evidence from the Clinic and Laboratory Models)

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12 pages, 2236 KiB

Open AccessArticle

Evaluating the Interaction of Emerging Diseases on White-Tailed Deer Populations Using an Agent-Based Modeling Approach

by Miranda Strasburg and Sonja Christensen

Pathogens 2024, 13(7), 545; https://doi.org/10.3390/pathogens13070545 - 28 Jun 2024

Viewed by 287

Abstract

Disease co-occurrence in wildlife populations is common yet understudied. In the case of disease-caused mortality, the mortality attributed to one disease has the potential to buffer populations against subsequent alternative disease outbreaks by reducing populations and thus contacts needed to sustain disease transmission. [...] Read more.

Disease co-occurrence in wildlife populations is common yet understudied. In the case of disease-caused mortality, the mortality attributed to one disease has the potential to buffer populations against subsequent alternative disease outbreaks by reducing populations and thus contacts needed to sustain disease transmission. However, substantial disease-driven population declines may also prevent populations from recovering, leading to localized extinctions. Hemorrhagic disease (HD), a vector-transmitted, viral disease in white-tailed deer (WTD), similar to chronic wasting disease (CWD), a prion disease, has increased in frequency and distribution in the United States. However, unlike CWD, which progresses slowly, HD can cause mortality only days after infection. Hemorrhagic disease outbreaks can result in substantial localized mortality events in WTD near vector habitats such as wetlands and may reduce local deer densities and consequent CWD transmission. The objective of our study was to evaluate the potential for HD outbreaks to buffer CWD risk where the diseases co-occur. Using an agent-based modeling approach, we found that frequent, intense HD outbreaks have the potential to mitigate CWD risk, especially if those outbreaks occur shortly after CWD introduction. However, HD outbreaks that do not result in substantial WTD mortality are unlikely to impact CWD or WTD population dynamics. Severe HD outbreaks may reduce CWD cases and could present an opportunity for managers to boost CWD control initiatives in a post-HD outbreak year. Full article

(This article belongs to the Special Issue Advances in Chronic Wasting Disease)

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20 pages, 1978 KiB

Open AccessArticle

Water Stress and Black Cutworm Feeding Modulate Plant Response in Maize Colonized by Metarhizium robertsii

by Imtiaz Ahmad, Maria del Mar Jimenez-Gasco and Mary E. Barbercheck

Pathogens 2024, 13(7), 544; https://doi.org/10.3390/pathogens13070544 - 27 Jun 2024

Viewed by 249

Abstract

Plants face many environmental challenges and have evolved different strategies to defend against stress. One strategy is the establishment of mutualistic associations with endophytic microorganisms which contribute to plant defense and promote plant growth. The fungal entomopathogen Metarhizium robertsii is also an endophyte [...] Read more.

Plants face many environmental challenges and have evolved different strategies to defend against stress. One strategy is the establishment of mutualistic associations with endophytic microorganisms which contribute to plant defense and promote plant growth. The fungal entomopathogen Metarhizium robertsii is also an endophyte that can provide plant-protective and growth-promoting benefits to the host plant. We conducted a greenhouse experiment in which we imposed stress from deficit and excess soil moisture and feeding by larval black cutworm (BCW), Agrotis ipsilon, to maize plants that were either inoculated or not inoculated with M. robertsii (Mr). We evaluated plant growth and defense indicators to determine the effects of the interaction between Mr, maize, BCW feeding, and water stress. There was a significant effect of water treatment, but no effect of Mr treatment, on plant chlorophyl, height, and dry biomass. There was no effect of water or Mr treatment on damage caused by BCW feeding. There was a significant effect of water treatment, but not Mr treatment, on the expression of bx7 and rip2 genes and on foliar content of abscisic acid (ABA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), and gibberellin 19 (GA19), whereas GA53 was modulated by Mr treatment. Foliar content of GA19 and cis-Zeatin (cZ) was modulated by BCW feeding. In a redundancy analysis, plant phenology, plant nutrient content, and foliar DIMBOA and ABA content were most closely associated with water treatments. This study contributes toward understanding the sophisticated stress response signaling and endophytic mutualisms in crops. Full article

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14 pages, 1907 KiB

Open AccessReview

Emerging and Re-Emerging Pathogens in Valvular Infective Endocarditis: A Review

by Maximilian Reisinger, Mateusz Kachel and Isaac George

Pathogens 2024, 13(7), 543; https://doi.org/10.3390/pathogens13070543 - 27 Jun 2024

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Abstract

Infective endocarditis (IE) is a microbial infection of the endocardial surface, most commonly affecting native and prosthetic valves of the heart. The epidemiology and etiology of the disease have evolved significantly over the last decades. With a growing elderly population, the incidence of [...] Read more.

Infective endocarditis (IE) is a microbial infection of the endocardial surface, most commonly affecting native and prosthetic valves of the heart. The epidemiology and etiology of the disease have evolved significantly over the last decades. With a growing elderly population, the incidence of degenerative valvopathies and the use of prosthetic heart valves have increased, becoming the most important predisposing risk factors. This change in the epidemiology has caused a shift in the underlying microbiology of the disease, with Staphylococci overtaking Streptococci as the main causative pathogens. Other rarer microbes, including Streptococcus agalactiae, Pseudomonas aeruginosa, Coxiella burnetti and Brucella, have also emerged or re-emerged. Valvular IE caused by these pathogens, especially Staphylococcus aureus, is often associated with a severe clinical course, leading to high rates of morbidity and mortality. Therefore, prompt diagnosis and management are crucial. Due to the high virulence of these pathogens and an increased incidence of antimicrobial resistances, surgical valve repair or replacement is often necessary. As the epidemiology and etiology of valvular IE continue to evolve, the diagnostic methods and therapies need to be progressively advanced to ensure satisfactory clinical outcomes. Full article

(This article belongs to the Special Issue Emerging and Re-emerging Bacterial Infections)

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18 pages, 1587 KiB

Open AccessArticle

Neurodevelopmental Outcomes of Preschoolers with Antenatal Zika Virus Exposure Born in the United States

by Sarah B. Mulkey, Elizabeth Corn, Meagan E. Williams, Emily Ansusinha, Robert H. Podolsky, Margarita Arroyave-Wessel, Gilbert Vezina, Colleen Peyton, Michael E. Msall and Roberta L. DeBiasi

Pathogens 2024, 13(7), 542; https://doi.org/10.3390/pathogens13070542 - 27 Jun 2024

Viewed by 183

Abstract

Neurodevelopmental outcomes for preschool-age children in the United States with in utero Zika virus (ZIKV) exposure have not yet been reported. We performed a case-control study to assess whether children exposed in utero to ZIKV have abnormal neurodevelopment at age 4–5 years compared [...] Read more.

Neurodevelopmental outcomes for preschool-age children in the United States with in utero Zika virus (ZIKV) exposure have not yet been reported. We performed a case-control study to assess whether children exposed in utero to ZIKV have abnormal neurodevelopment at age 4–5 years compared to unexposed controls. Thirteen ZIKV-exposed cases that did not have microcephaly or other specific features of congenital Zika syndrome and 12 controls were evaluated between ages 4–5 years. Child neurodevelopment was assessed using the Pediatric Evaluation of Disability Inventory, Behavior Rating Inventory of Executive Function, Peabody Picture Vocabulary Test, Bracken School Readiness Assessment (BSRA), and Movement Assessment Battery for Children (MABC). Caregivers answered questions on the child’s medical history and family demographics. Cases and controls were evaluated at mean (SD) ages 4.9 (0.3) and 4.8 (0.4) years, respectively. Caregivers reported more behavior and mood problems in cases than controls. MABC scores showed more gross and fine motor coordination difficulties among cases than controls. Controls trended towards higher performance on concepts underlying school readiness on BSRA. Three cases had a diagnosis of autism spectrum disorder or global developmental delay. Continued follow-up through school age for children with prenatal ZIKV exposure is needed to understand the impact of in utero ZIKV exposure on motor coordination, cognition, executive function, and academic achievement. Full article

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12 pages, 969 KiB

Open AccessArticle

Asymptomatic Malaria Reservoirs in Honduras: A Challenge for Elimination

by Sharon Banegas, Denis Escobar, Alejandra Pinto, Marcela Moncada, Gabriela Matamoros, Hugo O. Valdivia, Allan Reyes and Gustavo Fontecha

Pathogens 2024, 13(7), 541; https://doi.org/10.3390/pathogens13070541 - 27 Jun 2024

Viewed by 316

Abstract

Background: Efforts on a global scale for combating malaria have achieved substantial progress over the past twenty years. Two Central American nations have accomplished their goal of eliminating malaria: El Salvador and Belize. Honduras has decreased the incidence of malaria and now reports [...] Read more.

Background: Efforts on a global scale for combating malaria have achieved substantial progress over the past twenty years. Two Central American nations have accomplished their goal of eliminating malaria: El Salvador and Belize. Honduras has decreased the incidence of malaria and now reports fewer than 4000 malaria cases annually, aspiring to reach elimination by 2030. To accomplish this goal, it is essential to assess the existing strategies employed for malaria control and to address the task of incorporating novel intervention strategies to identify asymptomatic reservoirs. Methods: A survey for detecting asymptomatic cases was carried out in the community of Kaukira, in Gracias a Dios, Honduras, focusing on malaria transmission during 2023. Asymptomatic community members were recruited as participants, malaria screening was performed through a rapid diagnostic test in situ, and a blood sample was collected on filter paper. Highly sensitive molecular assays based on photo-induced electron transfer PCR (PET-PCR) were performed to detect the two species of Plasmodium circulating in Honduras: Plasmodium vivax and Plasmodium falciparum. In addition, the identification of the parasite species was verified by amplifying three genetic markers (Pvmsp3α, Pvmsp3ß, and Pfmsp1). Results: A total of 138 participants were recruited, mostly adult women. All individuals tested negative on the rapid diagnostic test. Positive results for malaria were detected by PET-PCR in 17 samples (12.3%). Most samples (12 out of 17) were amplified with a Ct value between 37 and 42, indicating very low parasitemias. Out of the 17 samples, 16 of them also showed amplification in the species assays. There were nine cases of P. falciparum infections and seven cases of P. vivax infections that were further confirmed by nested PCR (nPCR) of Pvmsp3 and Pfmsp1. Parasitemias ranged from 100 p/μL to less than 0.25 p/μL. One sample showed mixed infection. Conclusions: The existence of asymptomatic malaria reservoirs in Honduras can contribute to disease transmission and pose a challenge that may hinder elimination efforts, requiring public health authorities to modify surveillance strategies to identify the disease and treat this population accordingly. Full article

(This article belongs to the Special Issue Current Research on Host–Pathogen Interaction in 2024)

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10 pages, 246 KiB

Open AccessArticle

Persistently Elevated Expression of Systemic, Soluble Co-Inhibitory Immune Checkpoint Molecules in People Living with HIV before and One Year after Antiretroviral Therapy

by Robyn-Brooke Labuschagne Naidoo, Helen C. Steel, Annette J. Theron, Ronald Anderson, Gregory R. Tintinger and Theresa M. Rossouw

Pathogens 2024, 13(7), 540; https://doi.org/10.3390/pathogens13070540 - 27 Jun 2024

Viewed by 191

Abstract

Introduction: Increasing drug resistance and the absence of a cure necessitates exploration of novel treatment strategies for people living with HIV (PLWH). Targeting of soluble co-inhibitory immune checkpoint molecules (sICMs) represents a novel, potentially effective strategy in the management of HIV. Methods: In [...] Read more.

Introduction: Increasing drug resistance and the absence of a cure necessitates exploration of novel treatment strategies for people living with HIV (PLWH). Targeting of soluble co-inhibitory immune checkpoint molecules (sICMs) represents a novel, potentially effective strategy in the management of HIV. Methods: In this retrospective, longitudinal, observational study, the plasma levels of five prominent co-inhibitory sICMs—CTLA-4, LAG-3, PD-1 and its ligand PD-L1, as well as TIM-3—were quantified in 68 PLWH—before and one year after antiretroviral therapy (ART)—and compared with those of 15 healthy control participants. Results: Relative to control participants, PLWH had substantially elevated pre-treatment levels of all five co-inhibitory sICMs (p < 0.0001–p < 0.0657), which, over the 12-month period of ART, remained significantly higher than those of controls (p < 0.0367–p < 0.0001). PLWH with advanced disease, reflected by a CD4+ T cell count <200 cells/mm³ before ART, had the lowest levels of CTLA-4 and LAG-3, while participants with pre-treatment HIV viral loads ≥100,000 copies/mL had higher pre-treatment levels of TIM-3, which also persisted at 12 months. Conclusions: Plasma levels of CTLA-4, LAG-3, PD-1, PD-L1 and TIM-3 were significantly elevated in treatment-naïve PLWH and remained so following one year of virally-suppressive ART, possibly identifying LAG-3 and TIM-3 in particular as potential targets for adjuvant immunotherapy. Full article

(This article belongs to the Section Viral Pathogens)

13 pages, 1371 KiB

Open AccessArticle

Evaluation of Type I Interferon Treatment in Hospitalized COVID-19 Patients: A Retrospective Cohort Study

by Vivian Y. Tat, Pinghan Huang, Kamil Khanipov, Nathan Y. Tat, Chien-Te Kent Tseng and George Golovko

Pathogens 2024, 13(7), 539; https://doi.org/10.3390/pathogens13070539 - 26 Jun 2024

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Abstract

Coronavirus disease 2019 (COVID-19) continues to cause morbidity and mortality worldwide; therefore, effective treatments remain crucial to controlling it. As interferon-alpha (IFN-α) and -beta (β) have been proposed as COVID-19 treatments, we sought to assess their effectiveness on respiratory, cardiovascular, neurological, and psychiatric [...] Read more.

Coronavirus disease 2019 (COVID-19) continues to cause morbidity and mortality worldwide; therefore, effective treatments remain crucial to controlling it. As interferon-alpha (IFN-α) and -beta (β) have been proposed as COVID-19 treatments, we sought to assess their effectiveness on respiratory, cardiovascular, neurological, and psychiatric signs and symptoms, as well as PASC and death, in hospitalized COVID-19 patients without multiple sclerosis (MS). Using a federated data research network (TriNetX), we performed a retrospective cohort study of hospitalized COVID-19 patients without MS who received IFN-α or -β treatment, comparing them to a similar cohort who did not receive treatment. Following propensity-score matched analyses, we demonstrate that hospitalized COVID-19 patients who were treated with IFN-α or -β had significantly higher odds of death. In contrast, there was no significant difference in any other outcomes between 1–30 days or 1 day to anytime afterward. Overall, hospitalized COVID-19 patients without MS who were treated with IFN-α or -β had similar short- and long-term sequelae (except for mortality) as those who did not receive treatment. The potential benefits of utilizing IFN-α or -β treatment as therapeutics remain to be realized, and our research highlights the need to explore repurposing drugs for COVID-19 using real-world evidence. Full article

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11 pages, 2293 KiB

Open AccessArticle

The Prevalence of Antibiotic Tolerance in Neisseria gonorrhoeae Varies by Anatomical Site

by Margaux Balduck, Akim Strikker, Zina Gestels, Saïd Abdellati, Dorien Van den Bossche, Irith De Baetselier, Chris Kenyon and Sheeba Santhini Manoharan-Basil

Pathogens 2024, 13(7), 538; https://doi.org/10.3390/pathogens13070538 - 26 Jun 2024

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Abstract

Background: Tolerance enables bacteria to survive intermittent antibiotic exposure without an increase in antimicrobial susceptibility. In this study, we investigated the presence of tolerance to three antimicrobials, ceftriaxone, azithromycin and ciprofloxacin, in clinical isolates and the WHO (World Health Organization) reference panel of [...] Read more.

Background: Tolerance enables bacteria to survive intermittent antibiotic exposure without an increase in antimicrobial susceptibility. In this study, we investigated the presence of tolerance to three antimicrobials, ceftriaxone, azithromycin and ciprofloxacin, in clinical isolates and the WHO (World Health Organization) reference panel of Neisseria gonorrhoeae. Methods: We used the modified tolerance disk (TD test) to assess for tolerance to ceftriaxone, azithromycin and ciprofloxacin in 14 WHO reference strains and 62 N. gonorrhoeae clinical isolates—evenly divided between anorectal and urogenital infections. The isolates underwent a three-step incubation process wherein the isolates were exposed to an antibiotic disk for 20 h of incubation (Step I), followed by the replacement of the antibiotic disk with a nutrient disk for overnight incubation (Step II) and additional overnight incubation with extra nutrients (Step III). Results: A total of 4 of the 62 clinical anorectal isolates and none of the urogenital isolates exhibited tolerance to azithromycin (p = 0.033). Tolerance to ceftriaxone and ciprofloxacin was observed in eight and four isolates, respectively, with no difference between infection sites. Tolerance was also detected in 8 (K, M, N, O, P, U, V, W) out of the 14 WHO reference strains, with varying patterns of tolerance to ceftriaxone (n = 8), ciprofloxacin (n = 2) and azithromycin (n = 1). Conclusions: This study identified ceftriaxone, azithromycin and ciprofloxacin tolerance in clinical and WHO reference N. gonorrhoeae isolates. Azithromycin tolerance was more common in anorectal than urogenital infections. Full article

(This article belongs to the Section Bacterial Pathogens)

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13 pages, 1132 KiB

Open AccessArticle

Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

by Virginia Friedrichs, Darwin Reicks, Jeffrey J. Zimmerman, Eric A. Nelson, Carola Sauter-Louis, Martin Beer, Jane Christopher-Hennings and Sandra Blome

Pathogens 2024, 13(7), 537; https://doi.org/10.3390/pathogens13070537 - 25 Jun 2024

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Abstract

The rapid spread of African swine fever virus (ASFV), causing severe and often lethal disease in domestic pigs and Eurasian wild boar, continues to be a threat to pig populations and dependent industries. Despite scientific achievements that have deepened our understanding of ASFV [...] Read more.

The rapid spread of African swine fever virus (ASFV), causing severe and often lethal disease in domestic pigs and Eurasian wild boar, continues to be a threat to pig populations and dependent industries. Despite scientific achievements that have deepened our understanding of ASFV pathogenesis, alternative transmission routes for ASFV remain to be elucidated. We previously demonstrated the efficient transmission of ASFV from infected boars to naïve recipient gilts via artificial insemination, thereby highlighting the importance of surveillance of boar semen prior to its shipment. Since the accurate and reliable detection of even low amounts of ASFV in boar semen is key to disease prevention and control, we established a suitable diagnostic workflow to efficiently detect the ASFV genome in boar semen. Here, we assessed the sensitivity of various routine nucleic acid extraction kits as well as qPCR protocols in detecting the ASFV genome in the blood and semen of infected boars. The feasibility of the respective kits and methods for future use in boar studs was also considered. Variability in sensitivity mostly concerned samples with low to very low amounts of the ASFV genome. Ultimately, we defined a well-suited workflow for precisely detecting the ASFV genome in boar semen as early as 2 days post ASFV infection. Full article

(This article belongs to the Special Issue Emergence and Control of African Swine Fever)

15 pages, 527 KiB

Open AccessArticle

Testing Hepatitis E Seroprevalence among HIV-Infected Patients in Greece: THE SHIP Study

by Nikolina Antonopoulou, Georgios Schinas, Zoi Kotsiri, Olga Tsachouridou, Konstantinos Protopapas, Vasileios Petrakis, Emmanouil C. Petrakis, Despoina Papageorgiou, Dimosthenis Tzimotoudis, Simeon Metallidis, Antonios Papadopoulos, Markos Marangos, Emmanouil Barbounakis, Diamantis P. Kofteridis, Periklis Panagopoulos, Charalambos Gogos, Apostolos Vantarakis and Karolina Akinosoglou

Pathogens 2024, 13(7), 536; https://doi.org/10.3390/pathogens13070536 - 24 Jun 2024

Viewed by 290

Abstract

Hepatitis E virus (HEV) poses significant health concerns worldwide, particularly among people living with HIV (PLWHIV), due to an increased risk of chronic infection and progression to cirrhosis in individuals with low CD4 cell counts. This study aimed to investigate the prevalence, chronicity [...] Read more.

Hepatitis E virus (HEV) poses significant health concerns worldwide, particularly among people living with HIV (PLWHIV), due to an increased risk of chronic infection and progression to cirrhosis in individuals with low CD4 cell counts. This study aimed to investigate the prevalence, chronicity potential, and risk factors of HEV infection among PLWHIV in Greece, where data are currently absent. A synchronic multicentric study encompassing five major Greek university hospitals was executed over 24 months, recruiting 696 PLWHIV participants. The prevalence of HEV IgG antibodies was 16.5%, with 8.6% showing evidence of acute HEV infection (HEV IgM). Active viral replication (HEV RNA) was present in 2.3% of the study population. Longitudinal analysis revealed that of the 25 initially anti-HEV IgM-positive individuals, only 3 seroconverted to IgG positivity, and among those with prior HEV RNA positivity (16), none showed evidence of active replication in subsequent tests. Comparative subgroup analysis highlighted the lack of significant differences in HIV-related parameters between HEV seropositive and seronegative individuals. Laboratory evaluations generally showed no significant disparities across most parameters; however, a higher seropositivity for Hepatitis A was observed in the HEV-positive subgroup. Our findings highlight a considerable prevalence of HEV among PLWHIV in Greece, with no observed cases of chronicity. Full article

(This article belongs to the Special Issue Editorial Board Members’ Collection Series: HIV and Viral Co-infections)

14 pages, 886 KiB

Open AccessArticle

Plasma-Activated Tap Water with Oxidative Potential Has an Inactivating Effect on Microbiological Contaminants in Aqueous Suspensions

by Nahla C. Droste, Mareike Hummert, Paul Leenders, Alexander Mellmann, Karsten Becker and Thorsten Kuczius

Pathogens 2024, 13(7), 535; https://doi.org/10.3390/pathogens13070535 - 24 Jun 2024

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Abstract

Plasma-activated water (PAW) generated from tap water has gained attention as a disinfectant when used directly in its pure form. Little is known about the application of PAW for bacterial inactivation in aqueous environments because its use in fluids results in dilutions. We [...] Read more.

Plasma-activated water (PAW) generated from tap water has gained attention as a disinfectant when used directly in its pure form. Little is known about the application of PAW for bacterial inactivation in aqueous environments because its use in fluids results in dilutions. We investigated the effect of PAW in aqueous suspensions simulating such dilutions, and we focused on the minimal addition of PAW volumes to bacterial aqueous suspensions still resulting in high inactivation rates. The antimicrobial effect was highly dependent on the activation of PAW. An increase in activation power from 90 to 100 W resulted in a greater microbial reduction with an identical 10 min activation time. The susceptibility to PAW dilutions was analyzed in detail regarding nine Gram-negative species out of Enterobacterales and other waterborne microorganisms as well as four Gram-positive species present in two different matrices, in saline and in tap water, at high concentrations simulating massive contamination situations. For this purpose, the PAW activation setting of 90 W and 30 min was defined in order to be able to differentiate the limitations of inactivation in individual bacterial species. The Gram-negatives in saline demonstrated susceptibility when one volume unit of PAW was added. However, twice the PAW volume was necessary for inactivation when bacteria were present in tap water. Gram-positive microorganisms were more robust, indicated by prolonged contact times before inactivation. Our results indicate that PAW can be used for bacterial decontamination processes in aqueous environments when added in surplus. Optimized activation settings such as electric power to generate PAW and the contact times to the samples increase the effect of the inactivation a wide range of bacteria, regardless of their resistance profiles. Full article

(This article belongs to the Section Bacterial Pathogens)

10 pages, 575 KiB

Open AccessCommunication

Trueperella Pyogenes—Strain Diversity and Occurrence in Dairy Herds

by Nicole Wente, Stefanie Leimbach, Svenja Woudstra and Volker Krömker

Pathogens 2024, 13(7), 534; https://doi.org/10.3390/pathogens13070534 - 24 Jun 2024

Viewed by 344

Abstract

Trueperella (T.) pyogenes is a mastitis-causing pathogen formerly known to cause severe clinical mastitis (CM), especially during the summer, leading to milk losses and low recovery rates. Unfortunately, its transmission behavior within herds is unclear. The diversity and occurrence of T. pyogenes were [...] Read more.

Trueperella (T.) pyogenes is a mastitis-causing pathogen formerly known to cause severe clinical mastitis (CM), especially during the summer, leading to milk losses and low recovery rates. Unfortunately, its transmission behavior within herds is unclear. The diversity and occurrence of T. pyogenes were monitored to gain an initial insight into the infection transmission behavior of T. pyogenes in dairy herds and to lay a foundation for following targeted investigations. CM milk samples were collected from German herds, and one Swedish farm was sampled for isolates from subclinical mastitis. All in all, 151 T. pyogenes isolates from 16 herds were isolated, identified by MALDI TOF analysis and typed with RAPD PCR. Of these, 17 isolates originated from subclinical mastitis cases. We found that T. pyogenes mastitis occurred year-round, and clinical mastitis cases were caused by multiple strains (31 affected animals/28 strains). Instances of multiple cows being infected with the same T. pyogenes strain were rare and typically only involved a small number of animals at a time. However, if several quarters of a cow were affected, it was likely the same strain. Unlike clinical infections, subclinical T. pyogenes infections, in one investigated farm, harbored a dominant strain. Additionally, we found that T. pyogenes infections tended to persist and stay within a herd for a minimum of 7 months in the same or different cows. Full article

(This article belongs to the Special Issue Microbial Pathogenesis and Emerging Infections)

19 pages, 3407 KiB

Open AccessSystematic Review

Difference in the Intestinal Microbiota between Breastfeed Infants and Infants Fed with Artificial Milk: A Systematic Review

by Francesco Inchingolo, Angelo Michele Inchingolo, Giulia Latini, Laura Ferrante, Elisabetta de Ruvo, Merigrazia Campanelli, Marialuisa Longo, Andrea Palermo, Alessio Danilo Inchingolo and Gianna Dipalma

Pathogens 2024, 13(7), 533; https://doi.org/10.3390/pathogens13070533 - 24 Jun 2024

Viewed by 369

Abstract

The gut microbiota (GM) plays a crucial role in human health, particularly during the first years of life. Differences in GM between breastfed and formula (F)-fed infants may influence long-term health outcomes. This systematic review aims to compare the gut microbiota of breastfed [...] Read more.

The gut microbiota (GM) plays a crucial role in human health, particularly during the first years of life. Differences in GM between breastfed and formula (F)-fed infants may influence long-term health outcomes. This systematic review aims to compare the gut microbiota of breastfed infants with that of F-fed infants and to evaluate the clinical implications of these differences. We searched databases on Scopus, Web of Science, and Pubmed with the following keywords: “gut microbiota”, “gut microbiome”, and “neonatal milk”. The inclusion criteria were articles relating to the analysis of the intestinal microbiome of newborns in relation to the type of nutrition, clinical studies or case series, excluding reviews, meta-analyses, animal models, and in vitro studies. The screening phase ended with the selection of 13 publications for this work. Breastfed infants showed higher levels of beneficial bacteria such as Bifidobacterium and Lactobacillus, while F-fed infants had a higher prevalence of potentially pathogenic bacteria, including Clostridium difficile and Enterobacteriaceae. Infant feeding type influences the composition of oral GM significantly. Breastfeeding promotes a healthier and more diverse microbial ecosystem, which may offer protective health benefits. Future research should explore strategies to improve the GM of F-fed infants and understand the long-term health implications. Full article

(This article belongs to the Special Issue Oral Microbiome and Human Systemic Health)

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12 pages, 640 KiB

Open AccessArticle

Molecular Characterization and Antibacterial Resistance Determination of Escherichia coli Isolated from Fresh Raw Mussels and Ready-to-Eat Stuffed Mussels: A Major Public Health Concern

by Artun Yibar, Izzet B. Saticioglu, Nihed Ajmi and Muhammed Duman

Pathogens 2024, 13(7), 532; https://doi.org/10.3390/pathogens13070532 - 24 Jun 2024

Viewed by 346

Abstract

Our study focused exclusively on analyzing Escherichia coli (E. coli) contamination in fresh raw mussels and ready-to-eat (RTE) stuffed mussels obtained from authorized and regulated facilities. However, it is critical to recognize that such contamination represents a significant public health threat [...] Read more.

Our study focused exclusively on analyzing Escherichia coli (E. coli) contamination in fresh raw mussels and ready-to-eat (RTE) stuffed mussels obtained from authorized and regulated facilities. However, it is critical to recognize that such contamination represents a significant public health threat in regions where unauthorized harvesting and sales practices are prevalent. This study aimed to comprehensively assess the prevalence, molecular characteristics, and antibacterial resistance profiles of E. coli in fresh raw mussels and RTE stuffed mussels. E. coli counts in fresh raw mussel samples ranged from 1 to 2.89 log CFU/g before cooking, with a significant reduction observed post-cooking. RTE stuffed mussel samples predominantly exhibited negligible E. coli presence (<1 log CFU/g). A phylogenetic analysis revealed a dominance of phylogroup A, with variations in the distribution observed across different sampling months. Antibacterial resistance was prevalent among the E. coli isolates, notably showing resistance to ampicillin, streptomycin, and cefotaxime. Extended-spectrum β-lactamase (ESβL) production was rare, with only one positive isolate detected. A variety of antibacterial resistance genes, including tetB and sul1, were identified among the isolates. Notably, virulence factor genes associated with pathogenicity were absent. In light of these findings, it is imperative to maintain rigorous compliance with quality and safety standards at all stages of the mussel production process, encompassing harvesting, processing, cooking, and consumption. Continuous monitoring, implementation of rigorous hygiene protocols, and responsible antibacterial drug use are crucial measures in mitigating food safety risks and combating antibacterial resistance. Stakeholders, including seafood industry players, regulatory agencies, and healthcare professionals, are essential to ensure effective risk mitigation and safeguard public health in the context of seafood consumption. Full article

(This article belongs to the Special Issue Detection and Epidemiology of Drug-Resistant Bacteria)

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12 pages, 255 KiB

Open AccessArticle

Prevalence of Subgingival Aggregatibacter actinomycetemcomitans: Descriptive Cross-Sectional Study

by Nabil Khzam, Omar Kujan, Dorte Haubek, Aysen Arslan, Anders Johansson, Jan Oscarsson, Zeinab Razooqi and Leticia Algarves Miranda

Pathogens 2024, 13(7), 531; https://doi.org/10.3390/pathogens13070531 - 24 Jun 2024

Viewed by 373

Abstract

This paper aims to investigate the presence of Aggregatibacter actinomycetemcomitans and to assess potential indicators of the risk of severe form(s) of periodontitis. A descriptive cross-sectional study of 156 consecutive patients with periodontitis was conducted. Subgingival plaque samples were collected from the participants. [...] Read more.

This paper aims to investigate the presence of Aggregatibacter actinomycetemcomitans and to assess potential indicators of the risk of severe form(s) of periodontitis. A descriptive cross-sectional study of 156 consecutive patients with periodontitis was conducted. Subgingival plaque samples were collected from the participants. The identification of A. actinomycetemcomitans was performed using quantitative polymerase chain reaction. A descriptive analysis, a chi-square test, and a binary logistic regression statistical evaluation were performed. The prevalence of A. actinomycetemcomitans in this population of 156 participants was 17.30% (27 patients). The prevalence of stage-III periodontitis was 75.6% and greater in older men, while the prevalence of stage-IV periodontitis was 22.4% and greater in younger women. We observed a significant relation between the risk of severe periodontitis (stage-IV) and poor oral hygiene (p = 0.006), attendance at dental appointments (p ≤ 0.001), and familial history of periodontitis (p = 0.032). In conclusion, twenty-seven individuals were positive for A. actinomycetemcomitans. Poor oral hygiene, family history of periodontitis, and irregular attendance at dental appointments were identified as potential risk factors for severe periodontitis in this cohort. Full article

(This article belongs to the Special Issue Oral Microbiome and Human Systemic Health)

12 pages, 4524 KiB

Open AccessArticle

Invasive Fungal Infections of the Head and Neck: A Tertiary Hospital Experience

by Tieying Hou, W. Robert Bell and Hector Mesa

Pathogens 2024, 13(7), 530; https://doi.org/10.3390/pathogens13070530 - 23 Jun 2024

Viewed by 308

Abstract

From the existing millions of fungal species, only a few cause disease. In this study, we investigated invasive fungal infections in the head and neck (H&N) over a 19-year period (2005 to 2024) at a large academic healthcare system. Among the 413 documented [...] Read more.

From the existing millions of fungal species, only a few cause disease. In this study, we investigated invasive fungal infections in the head and neck (H&N) over a 19-year period (2005 to 2024) at a large academic healthcare system. Among the 413 documented fungal H&N infections, 336 were noninvasive, and 77 were invasive. The highest incidence of invasive infections occurred in the sinonasal cavities, with a 15-fold difference compared to other sites. Most infections affected adults over 40 years old. The most common organisms were Mucorales (51%), hyaline molds (29%), and Candida (11%). Risk factors included malignancy, transplant, diabetes, and illicit drug use. Mortality was high in patients with malignancy and/or transplant. Infections affecting the mandible were usually a complication of osteoradionecrosis and were associated with the coinfection of Candida and Actinomyces. At other sites, infections were rare and were usually the result of penetrating injuries or immunosuppression. Treatment typically involved a combination of antifungals and surgical procedures. Full article

(This article belongs to the Special Issue An Update on Fungal Infections)

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10 pages, 1122 KiB

Open AccessReview

Short Course Antibiotic Therapy for Catheter-Related Septic Thrombosis: “Caveat Emptor!”: Duration of Therapy Should Not Be Set a Priori

by Alberto Enrico Maraolo, Giancarlo Ceccarelli, Mario Venditti and Alessandra Oliva

Pathogens 2024, 13(7), 529; https://doi.org/10.3390/pathogens13070529 - 22 Jun 2024

Viewed by 434

Abstract

There is a growing body of evidence showing no significant difference in clinical outcomes in patients with uncomplicated Gram-negative bloodstream infections (BSIs) receiving 7 or 14 days of therapy. However, the scenario may differ when complicated forms of BSI, such as catheter-related BSIs [...] Read more.

There is a growing body of evidence showing no significant difference in clinical outcomes in patients with uncomplicated Gram-negative bloodstream infections (BSIs) receiving 7 or 14 days of therapy. However, the scenario may differ when complicated forms of BSI, such as catheter-related BSIs (CRBSIs) burdened by septic thrombosis (ST), are considered. A recent study showed that a short course of antimicrobial therapy (≤3 weeks) had similar outcomes to a prolonged course on CRBSI-ST. From this perspective, starting from the desirable goal of shortening the treatment duration, we discuss how the path to the correct diagnosis and management of CRBSI-ST may be paved with several challenges. Indeed, patients with ST due to Gram-negative bacteria display prolonged bacteremia despite an indolent clinical course, requiring an extended course of antibiotic treatment guided by negative FUBCs results, which should be considered the real driver of the decision-making process establishing the length of antibiotic therapy in CRBSI-ST. Shortening treatment of complicated CRBSIs burdened by ST is ambitious and advisable; however, a dynamic and tailored approach driven by a tangible outcome such as negative FUBCs rather than a fixed-duration paradigm should be implemented for the optimal antimicrobial duration. Full article

(This article belongs to the Section Epidemiology of Infectious Diseases)

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16 pages, 14810 KiB

Open AccessArticle

Upregulation of Neuroinflammation-Associated Genes in the Brain of SARS-CoV-2-Infected Mice

by Soo-Jin Oh, Pratima Kumari, Tabassum Tasnim Auroni, Shannon Stone, Heather Pathak, Amany Elsharkawy, Janhavi Prasad Natekar, Ok Sarah Shin and Mukesh Kumar

Pathogens 2024, 13(7), 528; https://doi.org/10.3390/pathogens13070528 - 22 Jun 2024

Viewed by 332

Abstract

Neurological manifestations are a significant complication of coronavirus disease 2019 (COVID-19), but the underlying mechanisms are yet to be understood. Recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-induced neuroinvasion and encephalitis were observed in K18-hACE2 mice, leading to mortality. Our goal in this [...] Read more.

Neurological manifestations are a significant complication of coronavirus disease 2019 (COVID-19), but the underlying mechanisms are yet to be understood. Recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-induced neuroinvasion and encephalitis were observed in K18-hACE2 mice, leading to mortality. Our goal in this study was to gain insights into the molecular pathogenesis of neurological manifestations in this mouse model. To analyze differentially expressed genes (DEGs) in the brains of mice following SARS-CoV-2 infection, we performed NanoString gene expression analysis using three individual animal samples at 1, 3, and 6 days post-infection. We identified the DEGs by comparing them to animals that were not infected with the virus. We found that genes upregulated at day 6 post-infection were mainly associated with Toll-like receptor (TLR) signaling, RIG-I-like receptor (RLR) signaling, and cell death pathways. However, downregulated genes were associated with neurodegeneration and synaptic signaling pathways. In correlation with gene expression profiles, a multiplexed immunoassay showed the upregulation of multiple cytokines and chemokines involved in inflammation and cell death in SARS-CoV-2-infected brains. Furthermore, the pathway analysis of DEGs indicated a possible link between TLR2-mediated signaling pathways and neuroinflammation, as well as pyroptosis and necroptosis in the brain. In conclusion, our work demonstrates neuroinflammation-associated gene expression profiles, which can provide key insight into the severe disease observed in COVID-19 patients. Full article

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9 pages, 817 KiB

Open AccessReview

Hepatitis C Virus and Molecular Mimicry

by Lynette Goh and Nanda Kerkar

Pathogens 2024, 13(7), 527; https://doi.org/10.3390/pathogens13070527 - 22 Jun 2024

Viewed by 273

Abstract

This review delves into the interactions between hepatitis C virus (HCV) and the host immune system, shedding light on how by using the mechanism of molecular mimicry, the virus strategically evades the immune system, resulting in a cascade of diverse complications. HCV, notorious [...] Read more.

This review delves into the interactions between hepatitis C virus (HCV) and the host immune system, shedding light on how by using the mechanism of molecular mimicry, the virus strategically evades the immune system, resulting in a cascade of diverse complications. HCV, notorious for its ability to persistently infect hepatocytes, employs molecular mimicry to resemble host proteins, thereby avoiding immune detection and mounting an effective defense. This mimicry also triggers systemic autoimmune responses that lead to various sequelae. The objective of this review is to comprehensively explore the role of HCV-induced molecular mimicry, which not only facilitates viral survival but is also instrumental in developing autoimmune and inflammatory disorders. By mimicking host proteins, HCV triggers an immune response that inadvertently attacks the host, fostering the development of autoimmune and other inflammatory disorders. Understanding the nuanced mechanisms of HCV-mediated molecular mimicry provides crucial insights into the multifaceted sequelae of viral infections on host immune responses. Unravelling these complexities is paramount for advancing therapeutic strategies that not only target the virus directly but also mitigate the secondary autoimmune and inflammatory complications induced by HCV. Full article

(This article belongs to the Special Issue Hepatitis C Virus Infection in Humans and Animals: Pathogenesis, Prevention and Treatment)

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10 pages, 296 KiB

Open AccessArticle

Evaluation of Three Different Selective Media for Enumeration of Clostridium perfringens in Untreated and Treated Wastewater

by A. J. Karon, Emily S. Bailey and Mark D. Sobsey

Pathogens 2024, 13(7), 526; https://doi.org/10.3390/pathogens13070526 - 21 Jun 2024

Viewed by 243

Abstract

Current and emerging legislation in North Carolina and other regions calls for the enumeration of Clostridium perfringens as a surrogate indicator for protozoan parasites in various types of waters. Past studies that have evaluated selective media for the detection of this bacterium have [...] Read more.

Current and emerging legislation in North Carolina and other regions calls for the enumeration of Clostridium perfringens as a surrogate indicator for protozoan parasites in various types of waters. Past studies that have evaluated selective media for the detection of this bacterium have provided limited, conflicting, and inconclusive results. In this study, membrane filtration was used to enumerate C. perfringens as culturable spores or total culturable cells in 19 samples of untreated and 25 samples of partially treated wastewaters on 3 candidate media, Tryptose Sulfite Cycloserine Agar (TSC), CP ChromoSelect Agar (CPCS), and membrane Clostridium perfringens Agar (m-CP) in parallel, and the results were compared. Presumptive isolates from each agar were further subjected to phenotypic confirmation tests for acid phosphatase production and stormy fermentation to further determine the performance of each agar. The CPCS agar was determined to have the highest enumerative capacity of total C. perfringens cells when compared to both TSC agar and m-CP agar (p-value < 0.05), but there was no significant difference in its ability to detect spores when compared to TSC agar (p-value > 0.05). The overall specificity of CPCS agar as determined by agreement of results from both confirmation tests was 0.81, while the specificity of TSC agar was only 0.28. Based on its performance, ease of preparation and use and consistency of colony characteristics, CPCS agar is recommended as the preferred medium for C. perfringens enumeration in wastewater. Full article

(This article belongs to the Collection Waterborne Pathogens: Epidemiology, Surveillance, Control and Risk)

14 pages, 1143 KiB

Open AccessArticle

Distribution and Pathogenicity Differentiation of Physiological Races of Verticillium dahliae from Cotton Stems in Western China

by Jianwei Zhang, Aerguli Jiamahate, Hui Feng, Tohir A. Bozorov, Dawei Zhang, Jianwei Guo, Honglan Yang and Daoyuan Zhang

Pathogens 2024, 13(7), 525; https://doi.org/10.3390/pathogens13070525 - 21 Jun 2024

Viewed by 222

Abstract

Verticillium wilt, caused by the pathogenic fungus Verticillium dahliae, has emerged as a severe threat to cotton globally. However, little is known about the genetic diversity of this pathogen in an infected single cotton plant. In this study, we isolated three new [...] Read more.

Verticillium wilt, caused by the pathogenic fungus Verticillium dahliae, has emerged as a severe threat to cotton globally. However, little is known about the genetic diversity of this pathogen in an infected single cotton plant. In this study, we isolated three new V. dahliae strains from the disease stems of Gossypium hirsutum from the cotton field in Western China and assessed their pathogenicity to the cotton cultivar Xinnongmian-1 and its two transgenic lines, as well as two laboratory strains, VD592 and VD991. These three new V. dahliae strains were identified using DNA barcodes of tryptophan synthase (TS), actin (ACT), elongation factor 1-α (EF), and glyceraldehyde-3-phosphate dehydrogenase (GPD). Moreover, the haplotype analysis revealed that the three new races had distinct haplotypes at the TS locus. Furthermore, the results of culture features and genetic diversity of ISSR (inter-simple sequence repeat) revealed that there were separate V. dahliae strains, which were strong defoliating pathotypes belonging to race 2 type, as determined by particular DNA marker recognition. The identified strains demonstrated varied levels of pathogenicity by leaf disc and entire plant inoculation methods. Conservatively, these strains showed some pathogenicity on cotton lines, but were less pathogenic than the reference strains. The findings revealed that several strong defoliating V. dahliae pathotypes coexist on the same cotton plant. It indicats the importance of regular monitoring as an early warning system, as well as the detection and reporting of virulent pathogen strains and their effects on crop response. Full article

26 pages, 1057 KiB

Open AccessReview

Advances in Laboratory Diagnosis of Coronavirus Infections in Cattle

by Shaun van den Hurk, Girija Regmi, Hemant K. Naikare and Binu T. Velayudhan

Pathogens 2024, 13(7), 524; https://doi.org/10.3390/pathogens13070524 - 21 Jun 2024

Viewed by 487

Abstract

Coronaviruses cause infections in humans and diverse species of animals and birds with a global distribution. Bovine coronavirus (BCoV) produces predominantly two forms of disease in cattle: a respiratory form and a gastrointestinal form. All age groups of cattle are affected by the [...] Read more.

Coronaviruses cause infections in humans and diverse species of animals and birds with a global distribution. Bovine coronavirus (BCoV) produces predominantly two forms of disease in cattle: a respiratory form and a gastrointestinal form. All age groups of cattle are affected by the respiratory form of coronavirus, whereas the gastroenteric form causes neonatal diarrhea or calf scours in young cattle and winter dysentery in adult cattle. The tremendous impacts of bovine respiratory disease and the associated losses are well-documented and underscore the importance of this pathogen. Beyond this, studies have demonstrated significant impacts on milk production associated with outbreaks of winter dysentery, with up to a 30% decrease in milk yield. In North America, BCoV was identified for the first time in 1972, and it continues to be a significant economic concern for the cattle industry. A number of conventional and molecular diagnostic assays are available for the detection of BCoV from clinical samples. Conventional assays for BCoV detection include virus isolation, which is challenging from clinical samples, electron microscopy, fluorescent antibody assays, and various immunoassays. Molecular tests are mainly based on nucleic acid detection and predominantly include conventional and real-time polymerase chain reaction (PCR) assays. Isothermal amplification assays and genome sequencing have gained increased interest in recent years for the detection, characterization, and identification of BCoV. It is believed that isothermal amplification assays, such as loop-mediated isothermal amplification and recombinase polymerase amplification, among others, could aid the development of barn-side point-of-care tests for BCoV. The present study reviewed the literature on coronavirus infections in cattle from the last three and a half decades and presents information mainly on the current and advancing diagnostics in addition to epidemiology, clinical presentations, and the impact of the disease on the cattle industry. Full article

(This article belongs to the Special Issue Diagnostics of Emerging and Re-emerging Pathogens)

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13 pages, 1607 KiB

Open AccessArticle

Humoral Immune Response in Immunized Sheep with Bovine Coronavirus Glycoproteins Delivered via an Adenoviral Vector

by Annamaria Pratelli, Paolo Capozza, Sergio Minesso, Maria Stella Lucente, Francesco Pellegrini, Maria Tempesta, Valentina Franceschi, Canio Buonavoglia and Gaetano Donofrio

Pathogens 2024, 13(7), 523; https://doi.org/10.3390/pathogens13070523 - 21 Jun 2024

Viewed by 229

Abstract

Bovine coronavirus (BCoV) is distributed globally and mainly causes different clinical manifestations: enteric diarrhea in calves, winter dysentery in adults, and respiratory symptoms in cattle of all ages. Low mortality and high morbidity are the hallmarks of BCoV infection, usually associated with substantial [...] Read more.

Bovine coronavirus (BCoV) is distributed globally and mainly causes different clinical manifestations: enteric diarrhea in calves, winter dysentery in adults, and respiratory symptoms in cattle of all ages. Low mortality and high morbidity are the hallmarks of BCoV infection, usually associated with substantial economic losses for the livestock industry. Vaccination, combined with the implementation of biosecurity measures, is the key strategy for the prevention of infections. This pilot study evaluates the immunogenicity of a recombinant vaccine containing two BCoV antigens (S and M) in sheep, compared to vaccines containing only the M or S protein. Three groups of sheep were inoculated intramuscularly at day 0 and day 21 with recombinant adenoviruses expressing BCoV S protein (AdV-BCoV-S), BCoV M protein (AdV-BCoV-M), or both proteins (AdV-BCoV-S + M). Serum antibodies were evaluated using immunofluorescence (IF) and serum neutralization (SN) tests. Moderate seroconversion was observed by day 21, but serum antibodies detected via SN increased from 1:27.5 (day 21) to 1:90 (day 28) in sheep inoculated with the recombinant AdV expressing both the S- and M-BCoV proteins. Based on the SN results, a repeated-measures ANOVA test indicated a more significant difference in immune response between the three groups (F = 20.47; p < 0.001). The experimental investigation produced satisfactory results, highlighting that the S + M recombinant vaccine was immunogenic, stimulating a valid immune response. Despite some inherent limitations, including a small sample size and the absence of challenge tests, the study demonstrated the efficacy of the immune response induced via the recombinant vaccine containing both S and M proteins compared to that induced via the individual proteins S or M. Full article

(This article belongs to the Section Vaccines and Therapeutic Developments)

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