Lactococcosis caused by Lactococcus garvieae is a bacterial infection affecting fish with a considerable economic impact. Recently, L. garvieae has established itself as an opportunistic pathogen in humans. The aim of the current study was to test classical and molecular-biological methods for the identification of L. garvieae and examine antimicrobial susceptibility and capsule production, an important virulence factor. Additionally, tests for differentiation from closely related species, as well as epidemiological typing, were performed. In a period of 18 years (2002–2019), 24 isolates presumptively identified as L. garvieae were collected from Oncorhynchus mykiss and Salmo salar fish obtained either from retail stores or fish farms. In order to confirm the species, optimized PCR-based protocols were used. As a result, 21 of the tested strains were proved to be L. garvieae (n = 21). The remaining three isolates were Lactococcus lactis, Streptococcus iniae, and Enterococcus faecalis. Epidemiological typing by randomly amplified polymorphic DNA was performed. Except for a single KG+ isolate, all other strains belonged to the European capsular serotype KG−. All L. garvieae isolates showed susceptibility to all tested antibiotics with the exception of clindamycin, which was a diagnostic sign. A thorough optimization of diagnostic methods is essential to determining the etiology of specific infections affecting the personnel at risk in fish farms, the food industry, or within the broader community.
The virus discovered in 2019 in the city of Wuhan, China, which was later identified as SARS-CoV-2 and which spread to the level of a pandemic, put diagnostic methods to the test. Early in the pandemic, we developed a nested PCR assay for the detection of SARS-CoV-2, which we validated and applied to detect the virus in feline samples. The present study describes the application of the nested PCR test in parallel with LAMP for the detection of the virus in 427 nasopharyngeal and oropharyngeal human samples taken between October 2020 and January 2022. Of the swabs tested, there were 43 positives, accounting for 10.1% of all samples tested, with the negatives numbering 382, i.e., 89.5%, and there were 2 (0.4%) invalid ones. The nPCR results confirmed those obtained by using LAMP, with results concordant in both methods. Nasal swabs tested using nPCR confirmed the results of oropharyngeal and nasopharyngeal swab samples tested using LAMP and nPCR. The focus of the discussion is on the two techniques: the actual practical application of the laboratory-developed assays and the diagnostic value of nasal samples. The nPCR used is a reliable and sensitive technique for the detection of SARS-CoV-2 in nasopharyngeal, oropharyngeal, and nasal swab samples. However, it has some disadvantages related to the duration of the entire process, as well as a risk of contamination. Experiments were performed to demonstrate the infectivity of the virus from the positive isolates in vitro. A discrepancy was reported between direct and indirect methods of testing the virus and accounting for its ability to cause infection in vitro.
Background and Objectives: This study aimed to detect and genotype human papillomavirus (HPV) in tissues from oral squamous cell carcinoma (OSCC), oral potentially malignant disorders (OPMD), and healthy individuals. Materials and Methods: The study included 60 patients (31 men and 29 women; median age 60 years, IQR 41.5–69.8) admitted to the Department of Dental, Oral and Maxillofacial Surgery, Medical University of Sofia. Patients were divided into three groups: healthy oral mucosa (n = 20), OPMD (n = 20), and OSCC (n = 20). HPV was tested using punch biopsies with nested PCR and chip technology. Results: Low-risk HPV was found in four (20%) healthy individuals (types 6/11, 43), seven (35%) OPMD patients (types 6/11, 42, 43), and eleven (55%) OSCC patients (types 6/11, 42, 43). Pairwise comparison showed a significant difference in HPV positivity between healthy individuals and OSCC patients (p = 0.022). Among all HPV-positive OPMDs, the virus was detected in two leukoplakia cases (28.6%), three lichen planus cases (42.9%), one lichenoid lesion case (14.3%), and one proliferative verrucous leukoplakia case (14.3%). According to binary logistic regression, OSCC patients were 4.9 times more likely to be HPV-positive compared to healthy individuals (p = 0.027). Conclusions: HPV infection may play a potential role in the pathogenesis of OPMD and OSCC.
Pregnancy is a special period characterized by changes in the immune system as well as alteration of the microbiota predisposing to opportunistic infections, including tonsillopharyngitis. Streptococcus anginosus has numerous virulence factors, and it is associated with a variety of diseases. Being a part of the normal oral microbiota, it can be overlooked as a causative agent of infections in the oral cavity. A clinical case of a 29‐year‐old pregnant female who was allergic to penicillins and presented with recurrent tonsillopharyngitis has been described. Previous microbiological examinations have not shown any etiologic cause of patient’s symptoms. Two species were isolated. S. anginosus and S. oralis were identified using various methods, including nucleotide sequencing. S. anginosus was defined as the causative agent. Both were susceptible to all tested antibiotics. S. anginosus was positive for the duplicated sagA gene, encoding streptolysin S, an important factor of virulence. Based on clinical symptoms, paraclinical tests, immune competence, and characteristics of the strain, a multifactorial evaluation is required to determine whether an opportunist is a causative agent. Moreover, treatment of pregnant patients, particularly with antibiotics, is limited, which leads to therapeutic difficulties.
SARS-CoV-2 is a zoonotic virus with a proven ability to infect various animal species, including domestic cats. In the post-pandemic period of COVID 19, limited data still exist on the clinical course, shedding of infectious virus and diagnostic features in cats. The aim of this study was to investigate the spread of SARS-CoV-2 in cats in 2023, the clinical manifestations of the infection, the diagnostic algorithm, including molecular detection of viral components, the differential diagnosis of co-infection with FHV, FCV, Mycoplasma spp. and Chlamydia felis, serology, and the isolation of infectious SARS-CoV-2. The immunomodulatory therapy in animals with a standalone SARS-CoV-2 infection was applied. The study included oropharyngeal, conjunctival and nasal swab samples from 102 domestic cats with clinical signs. Of them, 20.6% (21/102) were positive for SARS-CoV-2, with 16.67% (17/102) of the cats showing various variants of co-infection with FHV, FCV, Mycoplasma spp. and Chlamydia felis. Four of the cats had a standalone SARS-CoV-2 with mild clinical manifestations that included serous discharges from the eyes, without change in the general condition. The virus was isolated from these samples. These four cats and their owners were positive for antibodies to the virus, and the owners were PCR-negative. The treatment of SARS-CoV-2 infection included the preparations Viusid, RX immunosuport, Vetomun and Lisymun. This is one of the first post-pandemic studies covering FHV, FCV, Mycoplasma spp. and Chlamydia felis in domestic cats with SARS-CoV-2 infection and further expands on the essential main idea including the specified pathogens of interest.
SARS-CoV-2 is a zoonotic virus with a proven ability to infect various animal species, including domestic cats. In the post-pandemic period of COVID 19, there are still limited data on the clinical course, shedding of infectious virus and diagnostic features in cats. The aim of this study was to investigate the spread of SARS-CoV-2 in cats in 2023, the clinical manifestations of the infection, the diagnostic algorithm including molecular detection of viral components, differential diagnosis of co-infection with FHV, FCV, Mycoplasma spp. and Chlamydia felis, serology and isolation of infectious SARS-CoV-2. The immunomodulatory therapy in animals with a standalone SARS-CoV-2 infection was applied. The study included oropharyngeal, conjunctival and nasal swab samples from 102 domestic cats with clinical signs. Of them, 20.6% (21/102) were positive for SARS-CoV-2, with 16.67% (17/102) of the cats showing various variants of co-infection with FHV, FCV, Mycoplasma spp. and Chlamydia felis. Four of the cats had a standalone SARS-CoV-2 with mild clinical manifestations that included serous discharges from the eyes, without change in the general condition. The virus was isolated from these samples. These four cats and their owners were positive for antibodies to the virus, and the owners were PCR-negative. The treatment of SARS-CoV-2 infection included the preparations Viusid, RX immunosuport, Vetomun and Lisymun. This is one of the first post-pandemic study covering FHV, FCV, Mycoplasma spp. and Chlamydia felis in domestic cats with SARS-CoV-2 infection and further expands on the essential main idea including the specified pathogens of interest.