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10 pages, 381 KiB  
Communication
Multiwave Locked System Laser Treatment Reduces the Bacterial Load in the Gingival Sulcus of Dogs After Plaque Removal
by Ivana Pallante, Paolo Squarzoni, Elisa Mazzotta, Nicola Pozzato and Monica Monici
Vet. Sci. 2025, 12(8), 767; https://doi.org/10.3390/vetsci12080767 - 16 Aug 2025
Viewed by 253
Abstract
Periodontal disease is a prevalent condition in companion animals. It is crucial to prevent the plaque and bacteria on tooth surfaces to avoid gingivitis and the more harmful periodontitis. The aim of the present study was to assess the impact of MLS laser [...] Read more.
Periodontal disease is a prevalent condition in companion animals. It is crucial to prevent the plaque and bacteria on tooth surfaces to avoid gingivitis and the more harmful periodontitis. The aim of the present study was to assess the impact of MLS laser treatment on the bacterial load by analyzing the gingival swabs of a total of 16 owned dogs with no history of dental disease that were selected from a cohort of patients admitted for plaque removal and dental hygiene procedures. Following each dental hygiene session, each dog received a single MLS laser therapy treatment (808–905 nm diode laser, frequency 36 Hz, and fluence 0.16 J/cm2). Swabs were collected from the two upper premolars before and after the laser treatment. These were submitted for mesophilic bacteria counts, and microbiological analysis was conducted on 10 positive cultures to evaluate the changes in the oral bacterial microbiota. MLS laser therapy statistically significantly reduced the mesophilic bacteria count by 1 log, with higher pre-laser treatment counts (n = 0.44; 5.77 ± 0.21 95%CI) in comparison to post-laser treatment counts (n = 0.73; 4.80 ± 0.346 95%CI). The MLS laser therapy was found to reduce the bacterial load in 80% of the subjects (p < 0.05). No significant differences pre- and post-laser treatment were observed in the bacteria species isolated from the microbiological cultures. MLS laser therapy appears to be a useful, non-invasive method for reducing the bacterial load in the treatment and prophylaxis of periodontal disease in dogs. Full article
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11 pages, 1158 KiB  
Article
Can Oral Fluids Replace Nasal Swabs in Swine Influenza A Virus (swIAV) PCR Diagnostics?
by Aleksandra Woźniak, Piotr Cybulski, Pia Ryt-Hansen, Lars Erik Larsen, Kinga Biernacka, Dagmara Miłek and Tomasz Stadejek
Pathogens 2025, 14(8), 808; https://doi.org/10.3390/pathogens14080808 - 14 Aug 2025
Viewed by 253
Abstract
The diagnosis of swine influenza A virus (swIAV) has to involve laboratory detection, as the clinical signs are not pathognomonic. Nasal swabs (NSs) have been the preferred sample material for swIAV PCR diagnostics, but oral fluid (OF) is a convenient alternative material. In [...] Read more.
The diagnosis of swine influenza A virus (swIAV) has to involve laboratory detection, as the clinical signs are not pathognomonic. Nasal swabs (NSs) have been the preferred sample material for swIAV PCR diagnostics, but oral fluid (OF) is a convenient alternative material. In this study, NSs and OFs from 35 Polish swine herds were collected and tested with real-time RT-PCR in order to assess swIAV circulation patterns in Poland and improve protocols for efficient, non-invasive and cost-effective swIAV surveillance in pig farms. The study showed that the swIAV RNA was detected in 65.7% of the tested farms. In total, 21.2% of NS pools and 48.6% of OF samples were positive for swIAV. The Ct values in NS pools and OFs were similar (p > 0.05), but a significant reduction (p < 0.05) in swIAV prevalence in NSs was observed in nursery pigs from farms applying swIAV vaccinations. Successful subtyping was achieved more effectively with OFs compared to NSs, and the H1avN2 was most prevalent subtype detected. The results emphasized that OF can be useful for monitoring swIAV and subtyping. However, OFs cannot replace NSs, which were more useful in the assessment of the effect of swIAV vaccinations in nursery pigs. Full article
(This article belongs to the Special Issue Emergence and Re-Emergence of Animal Viral Diseases)
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13 pages, 255 KiB  
Article
Genetic, Psychological, and Behavioural Factors Associated with Subtypes of Pain-Related Temporomandibular Disorders
by Marko Zlendić, Ema Vrbanović Đuričić, Koraljka Gall Trošelj, Marko Tomljanović, Kristina Vuković Đerfi, Ivan Alajbeg and Iva Z. Alajbeg
Biomedicines 2025, 13(8), 1961; https://doi.org/10.3390/biomedicines13081961 - 12 Aug 2025
Viewed by 272
Abstract
Background: This genetic association study investigated single nucleotide polymorphism (SNP) in interleukin-8 (CXCL8) and opiorphin (OPRPN) genes, as well as psychological characteristics and oral behaviours, between patients with pain-related temporomandibular disorders (TMDp) and healthy controls. The aim was to [...] Read more.
Background: This genetic association study investigated single nucleotide polymorphism (SNP) in interleukin-8 (CXCL8) and opiorphin (OPRPN) genes, as well as psychological characteristics and oral behaviours, between patients with pain-related temporomandibular disorders (TMDp) and healthy controls. The aim was to examine associations and predictive value of these factors for TMDp subtypes: arthralgia and myalgia. Methods: A total of 85 patients with TMDp (arthralgia and/or myalgia) and 85 pain-free controls were included. Diagnoses were established following the Diagnostic Criteria for Temporomandibular Disorders (DC/TMD). All participants completed standardised self-report questionnaires assessing anxiety, depression, and oral behaviours. Buccal swabs were collected for DNA extraction, and SNP genotyping was performed using real-time PCR. Statistical analyses were conducted using dominant and recessive genetic models. Logistic regression models were applied to assess risk factors for each TMDp subtype. Results: Participants homozygous for the minor allele (CC genotype) of rs1387964 in OPRPN were significantly more prevalent in both arthralgia and myalgia groups compared to controls. Age and female sex predicted TMDp-arthralgia. Predictors of TMDp-myalgia included the CC genotype of rs1387964, age, female sex, anxiety, and depression. Conclusions: Genetic background and psychological characteristics were significant predictors of TMDp myalgia, highlighting a multifactorial profile for this TMDp subtype. Full article
(This article belongs to the Special Issue Biomarkers in Pain)
24 pages, 3149 KiB  
Article
Evaluation of Aggregate Oral Fluid Sampling for Early Detection of African Swine Fever Virus Infection
by Bonto Faburay, Kathleen O’Hara, Marta Remmenga, Theophilus Odoom, Sherry Johnson, William Tasiame, Matilda Ayim-Akonor, Benita Anderson, Kingsley Kwabena Amoako, Diane Holder, Wu Ping, Michelle Zajac, Vivian O’Donnell, Lizhe Xu, Robin Holland, Corrie Brown, Randall Levings and Suelee Robbe-Austerman
Viruses 2025, 17(8), 1089; https://doi.org/10.3390/v17081089 - 6 Aug 2025
Viewed by 572
Abstract
African swine fever (ASF) needs to be controlled, and prevention of the spread of African swine fever virus (ASFV) is dependent on enhanced surveillance and early disease detection. Commercial swine operations, especially in North America, Europe, and Asia, are characterized by comparatively large [...] Read more.
African swine fever (ASF) needs to be controlled, and prevention of the spread of African swine fever virus (ASFV) is dependent on enhanced surveillance and early disease detection. Commercial swine operations, especially in North America, Europe, and Asia, are characterized by comparatively large numbers of pigs, and sampling individual pigs, which represents the main strategy for current ASF surveillance, can be both costly and labor intensive. A study performed in Ghana was designed to estimate the diagnostic sensitivity of pen-based aggregate oral fluid testing for ASFV in infected pigs in a pen of 30 animals and to evaluate its utility as a tool to support surveillance of ASF in the US. This study was performed in three phases: (i) virus (Ghana ASFV24) amplification in a target host species to generate the challenge inoculum; (ii) titration of the inoculum (10% spleen homogenate) in target host species to determine the minimum dose inducing acute ASF in pigs with survival up to 5–6 days post-inoculation (dpi); and (iii) the main study, involving 186 pigs, consisting of 6 replicates of 30 pigs per pen and one seeder pig inoculated with wildtype ASFV (highly virulent genotype II) per pen. Daily sampling of aggregate oral fluids, uncoagulated blood, oropharyngeal swabs, fecal and water nipple swabs, and recording of rectal temperatures and clinical observations was carried out. The seeder pigs were each inoculated intramuscularly with 0.5 mL of the 10% spleen homogenate, which induced the desired clinical course of ASF in the pigs, with survival of up to 6 dpi. ASFV DNA was detected in the seeder pigs as early as 1 dpi and 2 dpi in the blood and oropharyngeal swabs, respectively. Transmission of ASFV from the seeder pigs to the contact pig population was detected via positive amplification of ASFV DNA in aggregate oral fluid samples at 3 days post-contact (dpc) in 4 out of 6 pens, and in all 6 pens, at 4 dpc. Testing of oropharyngeal swabs and blood samples from individual pigs revealed a variable number of ASFV-positive pigs between 3 and 5 dpc, with detection of 100% positivity between 6 and 18 dpc, the study endpoint. These findings demonstrate the potential utility of aggregate oral fluid sampling for sensitive and early detection of ASFV incursion into naïve swine herds. It also demonstrates that testing of environmental samples from the premises could further enhance overall ASF early detection and surveillance strategies. Full article
(This article belongs to the Collection African Swine Fever Virus (ASFV))
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11 pages, 217 KiB  
Article
Assessing Canine Parvovirus Vaccine Performance in Puppies with Maternally Derived Antibody: An Improved Study Design
by Jacqueline Pearce, Ellen Versmissen, David Sutton, Qi Cao and Ian Tarpey
Vaccines 2025, 13(8), 832; https://doi.org/10.3390/vaccines13080832 - 4 Aug 2025
Viewed by 578
Abstract
Background/Objectives: Typically, studies aiming to assess the ability of canine parvovirus (CPV) vaccines to immunise puppies with maternally derived antibody (MDA) are undertaken using group-housed puppies. Since live attenuated vaccine virus is invariably shed in the faeces, this can result in repeated [...] Read more.
Background/Objectives: Typically, studies aiming to assess the ability of canine parvovirus (CPV) vaccines to immunise puppies with maternally derived antibody (MDA) are undertaken using group-housed puppies. Since live attenuated vaccine virus is invariably shed in the faeces, this can result in repeated oral re-exposure and puppies which failed to respond to the initial vaccination may respond instead to shed vaccine virus in the environment, thus artificially enhancing the efficacy of the vaccine. This problem can be avoided by adopting a pair-housed study design where one vaccinated pup is housed with one unvaccinated sentinel. Using this design, we examine the capability of four commercially available canine parvovirus vaccines to immunise MDA-positive pups. Methods: Thirty-four 6-week-old puppies born to vaccinated dams were divided into four vaccine groups with similar MDA ranges. Within each group puppies were paired based on matching MDA titres, and each pair was housed in separate biocontainment accommodation. In each pair, the pup with the highest MDA was vaccinated and the other left as an unvaccinated sentinel. All vaccinates were given a single dose of one of the vaccines. Vaccinates and sentinels were then bled every 2–4 days and CPV antibody was measured. Daily rectal swabs were also collected from all pups to identify any shed vaccinal CPV. Results: All the pups vaccinated with Nobivac DP PLUS seroconverted, with significantly higher antibody titres compared to the pups in other vaccine groups, all shed vaccine virus, and all bar one of the sentinel pups seroconverted. In the other groups, only vaccinated pups with lower levels of MDA seroconverted and shed vaccine virus but none of the sentinel pups seroconverted. Conclusions: Different canine parvovirus vaccines differ in their ability to replicate in and immunise puppies with MDA, the levels of which may vary widely between individuals. The shedding of vaccinal CPV is an important consideration when designing studies to demonstrate efficacy in MDA-positive puppies. Full article
(This article belongs to the Section Veterinary Vaccines)
37 pages, 5366 KiB  
Article
Oral Microbiota Composition and Its Association with Gastrointestinal and Developmental Abnormalities in Children with Autism Spectrum Disorder
by Zuzanna Lewandowska-Pietruszka, Magdalena Figlerowicz and Katarzyna Mazur-Melewska
Microorganisms 2025, 13(8), 1822; https://doi.org/10.3390/microorganisms13081822 - 4 Aug 2025
Viewed by 533
Abstract
Autism Spectrum Disorder (ASD) is frequently accompanied by gastrointestinal disturbances, dietary selectivity, and altered stress responses, with growing evidence pointing to gut–brain axis involvement. While intestinal microbiota has been extensively studied, the role of the oral microbiota remains underexplored. This study investigates the [...] Read more.
Autism Spectrum Disorder (ASD) is frequently accompanied by gastrointestinal disturbances, dietary selectivity, and altered stress responses, with growing evidence pointing to gut–brain axis involvement. While intestinal microbiota has been extensively studied, the role of the oral microbiota remains underexplored. This study investigates the associations between oral microbiota composition and behavioral, gastrointestinal, dietary, and neuroendocrine parameters in children with ASD. A total of 45 children aged 2–18 years comprised the study group. Data collection included oral swabs for 16S rRNA gene sequencing, salivary cortisol sampling, dietary records, and standardized behavioral assessments using the Vineland Adaptive Behavior Scale. A total of 363 microbial species across 11 phyla were identified. Significant correlations were observed between specific bacterial taxa and functional gastrointestinal disorders (FGIDs), dietary patterns, salivary cortisol rhythms, and functioning. Children with FGIDs, food selectivity, or macronutrient imbalances exhibited enriched pro-inflammatory taxa (e.g., Selenomonas, Megasphaera), whereas those with typical cortisol secretion or higher adaptive functioning showed greater microbial diversity and abundance of health-associated genera (e.g., Bifidobacterium dentium). These findings suggest that oral microbiota profiles may reflect systemic physiological and neurobehavioral traits in children with ASD. Further longitudinal studies are needed to clarify causal relationships and support the development of microbiota-targeted interventions. Full article
(This article belongs to the Special Issue Focus on Pediatric Infectious Diseases)
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12 pages, 965 KiB  
Article
A Severe Form of Mpox Infection and the Current Epidemiological Status in Romania
by Anca Ruxandra Negru, David Valentin Mangaloiu, Ovidiu Vlaicu, Alexandra Cornovac, Violeta Molagic, Irina Duport-Dodot, Cătălin Tilișcan, Laurențiu Stratan, Adrian Marinescu, Lia Cavaropol, Mihaela Nicoleta Bercea, Andreea Marilena Păuna, Daniela Pițigoi, Victoria Aramă and Sorin-Stefan Aramă
Microorganisms 2025, 13(8), 1814; https://doi.org/10.3390/microorganisms13081814 - 3 Aug 2025
Viewed by 358
Abstract
Mpox has become a significant health concern since the global outbreak that began in 2022. The aim of this study is to present the epidemiological situation of Mpox in Romania during 2022–2023 and to describe a severe case of Mpox in a patient [...] Read more.
Mpox has become a significant health concern since the global outbreak that began in 2022. The aim of this study is to present the epidemiological situation of Mpox in Romania during 2022–2023 and to describe a severe case of Mpox in a patient who survived despite multiple co-pathologies. Forty-seven confirmed cases were reported at the national level, all in men, in 2022. The median age was 33 years. Twenty-six cases involved men who have sex with men (MSM), and twenty-three tested positive for HIV. We also describe a severe case involving a 34-year-old bisexual male with newly diagnosed AIDS who developed severe Mpox with persistent necrotic skin lesions, respiratory involvement, and multiple opportunistic infections: tuberculosis, pneumocystis pneumonia, syphilis, and oral candidiasis. The patient presented with fever, night sweats, weight loss, and dyspnea, with a single ulcerative facial lesion that later disseminated. Mpox infection was confirmed through PCR from skin lesion, serum, saliva, urine, rectal, nasal, and pharyngeal swab samples, with high viral loads persisting despite prolonged Tecovirimat therapy. The patient developed immune reconstitution inflammatory syndrome following the initiation of antiretroviral therapy. This case emphasizes the challenges of treating Mpox in immunocompromised patients. Full article
(This article belongs to the Special Issue Infectious Disease Surveillance in Romania)
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16 pages, 1622 KiB  
Article
Simian Foamy Virus Prevalence and Evolutionary Relationships in Two Free-Living Lion Tamarin Populations from Rio de Janeiro, Brazil
by Déa Luiza Girardi, Thamiris Santos Miranda, Matheus Augusto Calvano Cosentino, Caroline Carvalho de Sá, Talitha Mayumi Francisco, Bianca Cardozo Afonso, Flávio Landim Soffiati, Suelen Sanches Ferreira, Silvia Bahadian Moreira, Alcides Pissinatti, Carlos Ramon Ruiz-Miranda, Valéria Romano, Marcelo Alves Soares, Mirela D’arc and André Felipe Santos
Viruses 2025, 17(8), 1072; https://doi.org/10.3390/v17081072 - 31 Jul 2025
Viewed by 451
Abstract
Simian foamy virus (SFV) is a retrovirus that infects primates. However, epidemiological studies of SFV are often limited to captive populations. The southeastern Brazilian Atlantic Forest is home to both an endemic, endangered species, Leontopithecus rosalia, and an introduced species, Leontopithecus chrysomelas [...] Read more.
Simian foamy virus (SFV) is a retrovirus that infects primates. However, epidemiological studies of SFV are often limited to captive populations. The southeastern Brazilian Atlantic Forest is home to both an endemic, endangered species, Leontopithecus rosalia, and an introduced species, Leontopithecus chrysomelas, to which no data on SFV exist. In this study, we assessed the molecular prevalence of SFV, their viral load, and their phylogenetic relationship in these two species of primates. Genomic DNA was extracted from 48 oral swab samples of L. chrysomelas and 102 of L. rosalia. Quantitative PCR (qPCR) was performed to diagnose SFV infection and quantify viral load. SFV prevalence was found to be 23% in L. chrysomelas and 33% in L. rosalia. No age-related differences in prevalence were observed; however, L. rosalia showed a higher mean viral load (3.27 log10/106 cells) compared to L. chrysomelas (3.03 log10/106 cells). The polymerase gene sequence (213 pb) of L. rosalia (SFVlro) was clustered within a distinct SFV lineage found in L. chrysomelas. The estimated origin of SFVlro dated back approximately 0.0836 million years ago. Our study provides the first molecular prevalence data for SFV in free-living Leontopithecus populations while offering insights into the complex evolutionary history of SFV in American primates. Full article
(This article belongs to the Special Issue Spumaretroviruses: Research and Applications)
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14 pages, 3364 KiB  
Article
Microbial Load and Diversity of Bacteria in Wild Animal Carcasses Sold as Bushmeat in Ghana
by Daniel Oduro, Winnifred Offih-Kyei, Joanita Asirifi Yeboah, Rhoda Yeboah, Caleb Danso-Coffie, Emmanuel Boafo, Vida Yirenkyiwaa Adjei, Isaac Frimpong Aboagye and Gloria Ivy Mensah
Pathogens 2025, 14(8), 754; https://doi.org/10.3390/pathogens14080754 - 31 Jul 2025
Viewed by 350
Abstract
The demand for wild animal meat, popularly called “bushmeat”, serves as a driving force behind the emergence of infectious diseases, potentially transmitting a variety of pathogenic bacteria to humans through handling and consumption. This study investigated the microbial load and bacterial diversity in [...] Read more.
The demand for wild animal meat, popularly called “bushmeat”, serves as a driving force behind the emergence of infectious diseases, potentially transmitting a variety of pathogenic bacteria to humans through handling and consumption. This study investigated the microbial load and bacterial diversity in bushmeat sourced from a prominent bushmeat market in Kumasi, Ghana. Carcasses of 61 wild animals, including rodents (44), antelopes (14), and African civets (3), were sampled for microbiological analysis. These samples encompassed meat, intestines, and anal and oral swabs. The total aerobic bacteria plate count (TPC), Enterobacteriaceae count (EBC), and fungal counts were determined. Bacterial identification was conducted using MALDI-TOF biotyping. Fungal counts were the highest across all animal groups, with African civets having 11.8 ± 0.3 log10 CFU/g and 11.9 ± 0.2 log10 CFU/g in intestinal and meat samples, respectively. The highest total plate count (TPC) was observed in rodents, both in their intestines (10.9 ± 1.0 log10 CFU/g) and meat (10.9 ± 1.9 log10 CFU/g). In contrast, antelopes exhibited the lowest counts across all categories, particularly in EBC from intestinal samples (6.1 ± 1.5 log10 CFU/g) and meat samples (5.6 ± 1.2 log10 CFU/g). A comprehensive analysis yielded 524 bacterial isolates belonging to 20 genera, with Escherichia coli (18.1%) and Klebsiella spp. (15.5%) representing the most prevalent species. Notably, the detection of substantial microbial contamination in bushmeat underscores the imperative for a holistic One Health approach to enhance product quality and mitigate risks associated with its handling and consumption. Full article
(This article belongs to the Section Bacterial Pathogens)
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19 pages, 14428 KiB  
Article
Bivalent Oral Vaccine Using Attenuated Salmonella Gallinarum Delivering HA and NA-M2e Confers Dual Protection Against H9N2 Avian Influenza and Fowl Typhoid in Chickens
by Muhammad Bakhsh, Amal Senevirathne, Jamal Riaz, Jun Kwon, Ram Prasad Aganja, Jaime C. Cabarles, Sang-Ik Oh and John Hwa Lee
Vaccines 2025, 13(8), 790; https://doi.org/10.3390/vaccines13080790 - 25 Jul 2025
Viewed by 522
Abstract
Background: Fowl typhoid (FT), a septicemic infection caused by Salmonella Gallinarum (SG), and H9N2 avian influenza are two economically important diseases that significantly affect the global poultry industry. Methods: We exploited the live attenuated Salmonella Gallinarum (SG) mutant JOL3062 (SG: ∆lon [...] Read more.
Background: Fowl typhoid (FT), a septicemic infection caused by Salmonella Gallinarum (SG), and H9N2 avian influenza are two economically important diseases that significantly affect the global poultry industry. Methods: We exploited the live attenuated Salmonella Gallinarum (SG) mutant JOL3062 (SG: ∆lonpagLasd) as a delivery system for H9N2 antigens to induce an immunoprotective response against both H9N2 and FT. To enhance immune protection against H9N2, a prokaryotic and eukaryotic dual expression plasmid, pJHL270, was employed. The hemagglutinin (HA) consensus sequence from South Korean avian influenza A virus (AIV) was cloned under the Ptrc promoter for prokaryotic expression, and the B cell epitope of neuraminidase (NA) linked with matrix protein 2 (M2e) was placed for eukaryotic expression. In vitro and in vivo expressions of the H9N2 antigens were validated by qRT-PCR and Western blot, respectively. Results: Oral immunization with JOL3121 induced a significant increase in SG and H9N2-specific serum IgY and cloacal swab IgA antibodies, confirming humoral and mucosal immune responses. Furthermore, FACS analysis showed increased CD4+ and CD8+ T cell populations. On day 28 post-immunization, there was a substantial rise in the hemagglutination inhibition titer in the immunized birds, demonstrating neutralization capabilities of immunization. Both IFN-γ and IL-4 demonstrated a significant increase, indicating a balance of Th1 and Th2 responses. Intranasal challenge with the H9N2 Y280 strain resulted in minimal to no clinical signs with significantly lower lung viral titer in the JOL3121 group. Upon SG wildtype challenge, the immunized birds in the JOL3121 group yielded 20% mortality, while 80% mortality was recorded in the PBS control group. Additionally, bacterial load in the spleen and liver was significantly lower in the immunized birds. Conclusions: The current vaccine model, designed with a host-specific pathogen, SG, delivers a robust immune boost that could enhance dual protection against FT and H9N2 infection, both being significant diseases in poultry, as well as ensure public health. Full article
(This article belongs to the Special Issue Development of Vaccines Against Bacterial Infections)
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23 pages, 23270 KiB  
Article
Identifying Infectious Agents in Snakes (Boidae and Pythonidae) with and Without Respiratory Disease
by Marline M. Faulhaber, Florence Tardy, Anne V. Gautier-Bouchardon, Sabine Öfner, Sebastiaan Theuns, Sieglinde Coppens, Elisabeth Müller, Michael Pees and Rachel E. Marschang
Animals 2025, 15(15), 2187; https://doi.org/10.3390/ani15152187 - 25 Jul 2025
Viewed by 566
Abstract
Mycoplasmas are known respiratory pathogens in tortoises, but few studies exist in snakes. To better understand the correlation with clinical signs and co-infections, samples from mycoplasma-positive snakes with and without clinical respiratory disease were analyzed. Oral swabs from 15 snakes (pythons n = [...] Read more.
Mycoplasmas are known respiratory pathogens in tortoises, but few studies exist in snakes. To better understand the correlation with clinical signs and co-infections, samples from mycoplasma-positive snakes with and without clinical respiratory disease were analyzed. Oral swabs from 15 snakes (pythons n = 12, boas n = 3) were examined using polymerase chain reaction (PCR) and third-generation sequencing (TGS). Additionally, mycoplasma isolation assays were performed. Pathogens detected by PCR included Mycoplasmas (15/15, 100%), serpentoviruses (9/15, 60%), and Chlamydia sp. (2/15, 13%); those detected by TGS included Mycoplasmas (14/15, 93%), serpentoviruses (10/15, 67%), Chlamydia sp. (1/15, 7%), and 15 different bacterial species. Sequencing of the mycoplasma PCR products revealed a close genetic relationship to Mycoplasmopsis agassizii. TGS identified genetically distinct mycoplasmas and three different serpentoviruses. While mycoplasmas could not be successfully propagated, Brucella intermedia comb. nov. was identified in eight cultures. Respiratory disease in snakes is often multifactorial, involving various pathogens and environmental influences. This study demonstrates that comprehensive diagnostics are essential for understanding disease processes in snakes and improving the detection of diverse pathogens. Further research is needed to improve laboratory diagnostics for infectious diseases in reptiles and to better understand the roles of various pathogens in respiratory diseases in snakes. Full article
(This article belongs to the Special Issue Advances in Reptile and Amphibian Infectious Diseases)
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17 pages, 863 KiB  
Article
Porcine Sample Type Characteristics Associated with Sequencing and Isolation of Influenza A Virus
by Daniel C. A. Moraes, Onyekachukwu H. Osemeke, Michael A. Zeller, Amy L. Baker, Gustavo S. Silva, Giovani Trevisan, Daniel C. L. Linhares and Phillip C. Gauger
Vet. Sci. 2025, 12(7), 683; https://doi.org/10.3390/vetsci12070683 - 19 Jul 2025
Viewed by 608
Abstract
Understanding how sample type may influence the probability of influenza A virus (IAV) sequencing and isolation success can help improve the use of diagnostic tests and refine surveillance strategies in swine populations. The objective of this study was to evaluate the probability of [...] Read more.
Understanding how sample type may influence the probability of influenza A virus (IAV) sequencing and isolation success can help improve the use of diagnostic tests and refine surveillance strategies in swine populations. The objective of this study was to evaluate the probability of success for IAV hemagglutinin (HA) and neuraminidase (NA) Sanger sequencing and virus isolation in Madin–Darby Canine Kidney (MDCK) cells across different porcine sample types submitted to the Iowa State University Veterinary Diagnostic Laboratory (ISU VDL) from 2018 to 2024. Antemortem and postmortem sample types were selected and analyzed based on reverse transcription real-time polymerase chain reaction (RT-rtPCR) cycle threshold (Ct) values. The Ct values corresponding to 95%, 75%, and 50% probabilities of sequencing or virus isolation success were determined for each sample type. For antemortem samples, a 95% probability of success for HA Sanger sequencing on nasal swabs exhibited a Ct value of 27.8 from 1046 samples and 23.6 for NA sequencing based on 66 nasal swabs. Using oral fluids, HA and NA Sanger sequencing success was at Ct values of 27.3 from 3446 samples and 22.1 from 137 samples, respectively. For postmortem samples, lung tissue had the highest number of sequences for the HA and NA, with Ct values of 25.7 and 21.5, respectively. For a 95% probability of successful virus isolation, nasal swabs demonstrated a Ct value of 21.1 from 647 samples, while lungs had a Ct value of 18.7 from 5892 samples. This study determined that nasal swabs and lung tissue had the highest probability of IAV gene sequencing and virus isolation success, while oral fluids, a common swine diagnostic sample type that is easy to collect and welfare-friendly, can be effective for gene sequencing when using lower IAV RT-rtPCR Ct values, i.e., ≤27.3. These results provide practical expectations for successful IAV HA and NA gene sequencing and virus isolation at 95%, 75%, and 50% probabilities based on sample type and RT-rtPCR Ct values to improve diagnostic testing strategies in swine populations. Full article
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16 pages, 298 KiB  
Article
Antimicrobial-Resistant Staphylococcus spp. Harbored by Hedgehogs (Erinaceus europaeus) in Central Italy
by Fabrizio Bertelloni, Francesca Pauselli, Giulia Cagnoli, Roberto Biscontri, Renato Ceccherelli and Valentina Virginia Ebani
Antibiotics 2025, 14(7), 725; https://doi.org/10.3390/antibiotics14070725 - 18 Jul 2025
Viewed by 428
Abstract
Background/Objectives: European hedgehogs (Erinaceus europaeus) are present in areas where there is human activity; therefore, they can be a source of pathogens for other animals and humans. Methods: Eighteen hedgehog carcasses were collected and analyzed for Staphylococcus spp. Isolated strains were [...] Read more.
Background/Objectives: European hedgehogs (Erinaceus europaeus) are present in areas where there is human activity; therefore, they can be a source of pathogens for other animals and humans. Methods: Eighteen hedgehog carcasses were collected and analyzed for Staphylococcus spp. Isolated strains were typed and analyzed for exfoliative toxins genes and the phenotypic and genotypic characteristics of antimicrobial resistance. Results: A total of 54 strains were isolated and typed as S. aureus, S. xylosus, S. sciuri, S. pseudintermedius, S. simulans, S. chromogenes, S. epidermidis, S. hyicus, and S. lentus. No strains had the eta and etb genes coding for exfoliative toxins. Overall, 39/54 (72.20%) isolates showed phenotypic resistance to at least one antimicrobial and 21/54 (38.80%) showed more than one resistance. The lowest efficacy was observed for erythromycin, with 40/54 (74.08%) strains classified as intermediate and 6/54 (11.11%) classified as resistant. Among the 29 isolates shown to be penicillin-resistant, 11 (37.93%) were oxacillin-resistant, with a minimum inhibitory concentration (MIC). Among the 54 staphylococcal strains, 2 (3.70%) were resistant to vancomycin, both with an MIC value equal to the maximum concentration of the antibiotic tested (256 μg/mL) and 2 (3.70%) had an intermediate resistance profile with an 8 μg/mL MIC value. No strains had the genes vanA and vanB. Two of the 29 (6.90%) penicillin-resistant strains had the blaZ gene; 8 (27.13%) strains had the mecA gene. Overall, 2/54 (3.70%) isolates were classified as extensively drug-resistant (XDR) and 9/54 (16.66%) were classified as multidrug-resistant (MDR). Conclusions: Hedgehogs can harbor antimicrobial-resistant staphylococci and can be sources of these bacteria for other animals and humans. They can also serve as bioindicators of the pathogens and antimicrobial-resistant bacteria circulating in a given habitat. Full article
11 pages, 515 KiB  
Article
Systemic Antifolate Chemotherapy Does Not Select for Fluconazole-Resistant Candida: A Multicenter Clinical Study
by Dawid Żyrek, Joanna Nowicka, Magdalena Pajączkowska, Mariola Paściak, Katarzyna Machnik, Tomasz Werner, Zygmunt Konieczny, Piotr Jędrzejczak, Dominika Raźniewska, Gabriela Fijałkowska, Michał Piątek, Barbara Radecka, Kinga Żyrek, Elżbieta Woźniak-Grygiel and Iwona Dzieńdziora-Urbińska
Pathogens 2025, 14(6), 574; https://doi.org/10.3390/pathogens14060574 - 7 Jun 2025
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Abstract
Previous studies have demonstrated that Candida spp. isolates exposed in vitro to the folic acid antagonist methotrexate may develop multidrug cross-resistance to azole antifungals. The aim of this study was to determine whether systemic therapy with antineoplastic antifolates—pemetrexed or methotrexate—constitutes a risk factor [...] Read more.
Previous studies have demonstrated that Candida spp. isolates exposed in vitro to the folic acid antagonist methotrexate may develop multidrug cross-resistance to azole antifungals. The aim of this study was to determine whether systemic therapy with antineoplastic antifolates—pemetrexed or methotrexate—constitutes a risk factor for colonization or infection with fluconazole-resistant yeasts. The study group comprised 44 cancer patients who received high-dose systemic antifolate therapy, while the control group consisted of 48 patients without prior exposure to either methotrexate or pemetrexed. Oral swabs and relevant clinical data were collected from all participants. In total, 109 fungal strains representing 13 species were isolated, identified, and subsequently tested for fluconazole susceptibility. Fluconazole-resistant isolates were identified in 4 out of 44 (9.1%) antifolate-treated patients and in 3 out of 48 (6.3%) control patients. Our findings suggest that, although this phenomenon occurs in vitro, systemic antineoplastic antifolate therapy does not induce azole resistance among endogenous yeast species in vivo. Full article
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12 pages, 828 KiB  
Article
Molecular Prevalence of Leishmania infantum Infection from Oral Swabs Collected from Dogs in Region of Northwestern Spain
by Javier Merino-Goyenechea, Elora Valderas-García, Verónica Castilla Gómez de Agüero, Rafael Balaña-Fouce and María Martínez-Valladares
Pathogens 2025, 14(6), 569; https://doi.org/10.3390/pathogens14060569 - 6 Jun 2025
Viewed by 812
Abstract
Leishmaniasis is a serious zoonotic parasitic disease caused by the protist Leishmania infantum and transmitted by phlebotomine sandflies in the countries of the Mediterranean basin. Dogs are the species most susceptible to the disease and serve as a reservoir for transmission to humans, [...] Read more.
Leishmaniasis is a serious zoonotic parasitic disease caused by the protist Leishmania infantum and transmitted by phlebotomine sandflies in the countries of the Mediterranean basin. Dogs are the species most susceptible to the disease and serve as a reservoir for transmission to humans, making the Iberian Peninsula an endemic region for this infection. Although the regions close to the Mediterranean coast are the most prevalent regions of leishmaniasis in Spain, climatic factors are favouring the expansion of the vectors to more northern latitudes, where the disease was hardly known decades ago. This paper presents a prevalence study of L. infantum infection in the province of Zamora (northwestern Spain) using a non-invasive sample from canine buccal swabs and an innovative qPCR method to determine the etiologic agent. The parasite load of 151 randomly selected dogs from different points of the province was analysed during the period 2021–2022, with an estimated prevalence of 30%. In addition, the most common clinical signs of leishmaniasis in the dogs are described, and intrinsic factors associated with the nature of the dogs—such as sex, size, age as well as other factors related to the habitat in which they live and their geographical location—which could favour the disease, are evaluated. Full article
(This article belongs to the Section Parasitic Pathogens)
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