Apparent stability masks underlying change in a mule deer herd with unmanaged chronic wasting disease

The contagious prion disease “chronic wasting disease” (CWD) infects mule deer (Odocoileus hemionus) and related species. Unchecked epidemics raise ecological, socioeconomic, and public health concerns. Prion infection shortens a deer’s lifespan, and when prevalence (proportion of adults infected) becomes sufficiently high CWD can affect herd dynamics. Understanding population responses over time is key to forecasting long-term impacts. Here we describe unexpected stability in prevalence and abundance in a mule deer herd where CWD has been left unmanaged. High apparent prevalence (~30%) since at least 2005 likely drove observed changes in the proportion and age distribution of wild-type native prion protein (PRNP) gene homozygotes among deer sampled. Predation by mountain lions (Puma concolor) may be helping keep CWD in check. Despite stable appearances, prion disease nonetheless impairs adult survival and likely resilience in this deer herd, limiting its potential for growth despite refuge from hunter harvest and favorable habitat and winter conditions.

Immunofluorescence Targeting PBP2a Protein: A New Potential Methicillin Resistance Screening Test

The indiscriminate use of first-line drugs contributed to the spread of resistant bacteria, a major concern for both human and veterinary medicine. Methicillin resistance is acquired through the mecA gene, which encodes for the PBP2a protein and lends the resistance to β-lactams. Verifying the correspondence between gene harboring and protein expression and accelerating methicillin resistance diagnosis is critical to improve the management of antimicrobial administration and to reduce the spread of drug resistances. We tested the applicability of immunofluorescence targeting PBP2a protein to identify a new potential methicillin resistance screening test, ancillary to conventional culture methods. We collected 26 clinical Staphylococcus pseudintermedius (SP) isolates: 25 from canine pyoderma and 1 from dermatitis in a dog owner. SP is one of the most important etiological agents in canine pyoderma and can harbor the mecA gene. We performed PCR for mecA gene detection, broth microdilution (BMD) for phenotypic methicillin resistance, and immunofluorescence targeting PBP2a protein. Compared to the PCR as the gold standard, immunofluorescence showed an apparent prevalence of 34.6% vs. a true prevalence of 53.8%, with 100% specificity, 64.3% sensitivity, and 80.8% diagnostic accuracy. PBP2a expression showed isolate-dependent variability: in some isolates, most of the bacterial cells showed an intense and clearly membranous pattern, while in others only a few of them could be detected. Performing the assay in duplicate improved the diagnostic accuracy. Since the mecA gene is shared among the members of the Staphylococcus genus, the test can be applied to identify methicillin resistance independently from the staphylococcal species, both in human and animal samples. Being a rapid and easy method and providing the unique possibility to study the expression of PBP2a by directly visualizing the morphology, it could represent a new interesting tool for both research and diagnostics. To accelerate methicillin resistance diagnosis, it would be worth further testing of its performance on cytological samples.

The SARS-CoV-2 infection among students in the University of Porto: a cross-sectional study

Introduction: Incidence based on notified cases of SARS-CoV-2 infection underestimates the real extension of the infection. We aimed to quantify SARS-CoV-2 specific antibodies seroprevalence among University students in Porto. Methods: A rapid point of care testing for SARS-CoV-2 specific immunoglobulin (Ig) M and IgG antibodies was performed, and a questionnaire was applied to the 6512 voluntary students from September to December 2020. We computed the apparent IgM, IgG and IgM or IgG prevalence, and the true prevalence and 95% credible intervals (95% CI) using Bayesian inference. Results: We found an apparent prevalence (IgM or IgG) of 9.7%, the true prevalence being 7.9% (95% CI 4.9-11.1). Prevalence was significantly higher among males (10.9% vs 9.2%), international students (18.1% vs 10.4% local vs 8.8% nationally displaced) and increased with age. Those with a known risk contact, that experienced quarantine, had symptoms, or a previous negative molecular test had a higher seroprevalence. Of the 91 (1.4%) students who reported a molecular diagnosis, 86.7% were reactive for IgM or IgG. Conclusion: Based on immunological evidence infection was 5.6 times more frequent than if based on a molecular diagnosis. The higher seroprevalence among male, older, and international students emphasizes the importance of identifying particular groups.

Prevalence of SARS-CoV-2 antibodies among workers of the public higher education institutions of Porto, Portugal: a cross-sectional study

ObjectivesTo assess the prevalence of SARS-CoV-2-specific IgM and IgG antibodies among workers of the three public higher education institutions of Porto, Portugal, up to July 2020.MethodsA rapid point-of-care test for specific IgM and IgG antibodies of SARS-CoV-2 was offered to all workers (SD Biosensor STANDARD Q COVID-19 IgM/IgG Duo and STANDARD Q COVID-19 IgM/IgG Combo). Testing was performed and a questionnaire was completed by 4592 workers on a voluntary basis from 21 May to 31 July 2020. We computed the apparent IgM, IgG, and combined IgM or IgG prevalence, along with the true prevalence and 95% credible intervals (95% CrI) using Bayesian inference.ResultsWe found an apparent prevalence of 3.1% for IgM, 1.0% for IgG and 3.9% for either. The estimated true prevalence was 2.0% (95% CrI 0.1% to 4.3%) for IgM, 0.6% (95% CrI 0.0% to 1.3%) for IgG, and 2.5% (95% CrI 0.1% to 5.3%) for IgM or IgG. A SARS-CoV-2 molecular diagnosis was reported by 21 (0.5%) workers; and of these, 90.5% had a reactive IgG result. Seroprevalence was higher among those reporting contacts with confirmed cases, having been quarantined, having a previous molecular negative test or having had symptoms.ConclusionsThe seroprevalence among workers from the three public higher education institutions of Porto after the first wave of the SARS-CoV-2 infection was similar to national estimates for the same age working population. However, the estimated true seroprevalence was approximately five times higher than the reported SARS-CoV-2 infection based on a molecular test.

Detection of a novel enterotropic Mycoplasma gallisepticum-like in European Starling (Sturnus vulgaris) around poultry farms in France

Recent outbreaks of highly pathogenic avian influenza in southwest France have raised questions regarding the role of commensal wild birds in the introduction and dissemination of pathogens between poultry farms. To assess possible infectious contacts at the wild-domestic bird interface, the presence of Mycoplasma gallisepticum (MG) was studied in the two sympatric compartments in southwest France. Among various peridomestic wild birds (n=385), standard PCR primers targeting the 16S rRNA of MG showed a high apparent prevalence (up to 45%) in cloacal swabs of European starlings (Sturnus vulgaris, n=108), while the MG-specific mgc2 gene was not detected. No tracheal swab of these birds tested positive, and no clinical sign was observed in positive birds, suggesting commensalism in the digestive tract of starlings. A mycoplasma strain was then isolated from a starling swab and its whole genome was sequenced using both Illumina and Nanopore technologies. Phylogenetic analysis showed that it was closely related to MG and M. tullyi, although it was a distinct species. A pair of specific PCR primers targeting the mgc2-like gene of this MG-like strain was designed and used to screen again the same avian populations and a wintering urban population of starlings (n=50). Previous PCR results obtained in starlings were confirmed to be mostly due to this strain (20/22 positive pools). In contrast, the strain was not detected in fresh faeces of urban starlings. Furthermore, it was detected in one tracheal pool of cattle egrets and one cloacal pool of white wagtails, suggesting infectious transmissions between synanthropic birds with similar feeding behaviour. As the new starling mycoplasma was not detected in free-range ducks (n=80) in close contact with positive starlings, nor in backyard (n=320) and free-range commercial (n=720) chickens of the area, it might not infect poultry. However, it could be involved in mycoplasma gene transfer in such multi-species contexts.

Effect of vaccination on the apparent prevalence of bovine brucellosis in the state of Tocantins, Brazil

A cross-sectional study on the epidemiological situation of bovine brucellosis was carried out in the state of Tocantins to evaluate the effectiveness of its vaccination program. The state was divided into five regions, and a predetermined number of farms was randomly selected in each one. Females aged 24 months or older were randomly selected in each farm and diagnosed with brucellosis by serial serology (AAT and 2-ME). A total of 6,846 animals from 756 farms were examined. The prevalence of seropositive herds in the state was 6.42% [CI95%: 4.76-8.62], and the prevalence of seropositive animals was 2.21% [CI95%: 1.05-4.01]. The prevalence of seropositive herds was homogeneously distributed among regions. The 2002/2003 study estimated the prevalence of seropositive herds in the state to be 21.22% [CI95%: 19.33-23.11]. In conclusion, the vaccination program implemented in Tocantins, reaching vaccination coverage above 70% as of 2010, significantly reduced the prevalence of seropositive herds. Thus, continuing the vaccination program in the state is recommended, preferably increasing the quality of the processes involved, from commercialization to inoculation in animals, since immunization remains the most effective means to reduce the prevalence of brucellosis. In addition, animal replacement remains a major risk factor for bovine brucellosis in Tocantins since 20022003; therefore, the state must implement a strong health education program explaining to farmers the importance of testing animals for brucellosis before introducing them into their herds.

The GAR/RGG motif defines a family of nuclear alarmins

The nucleus is the target of autoantibodies in many diseases, which suggests intrinsic nuclear adjuvants that confer its high autoimmunogenicity. Nucleolin (NCL) is one abundant nucleolar autoantigen in systemic lupus erythematosus (SLE) patients and, in lupus-prone mice, it elicits autoantibodies early. With purified NCL, we observed that it was a potent alarmin that activated monocytes, macrophages and dendritic cells and it was a ligand for TLR2 and TLR4. NCL released by necrotic cells also exhibited alarmin activity. The NCL alarmin activity resides in its glycine/arginine-rich (GAR/RGG) motif and can be displayed by synthetic GAR/RGG peptides. Two more GAR/RGG-containing nucleolar proteins, fibrillarin (FBRL) and GAR1, were also confirmed to be novel alarmins. Therefore, the GAR/RGG alarmin motif predicts a family of nucleolar alarmins. The apparent prevalence of nucleolar alarmins suggests their positive contribution to tissue homeostasis by inducing self-limiting tissue inflammation with autoimmunity only occurring when surveillance is broken down.

The Occurrence of Mycobacterium avium Subspecies paratuberculosis Positive Milk Antibody ELISA Results in Dairy Cattle under Varying Time Periods after Skin Testing for Bovine Tuberculosis

Enzyme-linked immunosorbent assays (ELISA) are used to screen cows for Mycobacterium avium subspecies paratuberculosis (MAP) infections, informing Johne’s disease (JD) management practices in dairy herds. The causative agent of bovine tuberculosis (bTB), Mycobacterium bovis, and MAP share multiple antigens. Moreover, Mycobacterium avium subspecies avium is used in the single intradermal cervical comparative tests (SICCT) that are routinely used in early detection of cows infected with bTB. Although these are different types of immune responses, potentially the SICCT may interfere with the levels of MAP antibodies. This study aimed to clarify the relationship between the SICCT-MAP milk ELISA testing interval and apparent prevalence of JD risk statuses. Data from 51 herds were used, totalling 46,738 cow observations. The Poisson models showed that MAP milk ELISA testing at 14 day intervals post-SICCT statistically significantly increased the odds of detecting JD-positive cows compared to JD testing 85+ days post-SICCT. The odds ratio (OR) started at 2.5 in the first 14 day interval post-SICCT, increasing each two-week period to an OR of 4.0 at 57–70 days, to subsequently drop. Additionally, a herd history of bTB increased the odds of detecting JD-positive cows (OR = 1.2); this was relatively limited compared to the magnitude of the post-SICCT effect.

Prevalence of SARS-CoV-2 antibodies among workers of the public higher education institutions of Porto, Portugal

ObjectivesTo assess the prevalence of SARS-CoV-2 specific immunoglobulin (Ig) M and IgG antibodies among workers of the three public higher education institutions of Porto, Portugal, up to July 2020.MethodsA rapid point of care test for specific IgM and IgG antibodies of SARS-CoV-2 was offered to all workers. Testing was performed to and a questionnaire was completed by 4592 workers on a voluntary basis. We computed the apparent IgM, IgG, and combined IgM or IgG prevalence, along with the true prevalence and 95% credible intervals (95% CI) using Bayesian inference.ResultsWe found an apparent prevalence of 3.1% for IgM, 1.0% for IgG, and 3.9% for either antibody class. The estimated true prevalence was 2.0% (95% CI 0.1-4.3) for IgM, 0.6% (95% CI 0.0-1.3) for IgG and 2.5% (95% CI 0.1-5.3) for IgM or IgG. A SARS-CoV-2 molecular diagnosis was reported by 21 (0.5%) workers, and of these, 90.5% had a reactive IgG result. Seroprevalence was higher among those reporting known contacts with confirmed cases, having been quarantined, having a previous molecular negative test, or having had symptoms.ConclusionsThe seroprevalence among workers from the three public higher education institutions of Porto after the first wave of the SARS-CoV-2 infection was relatively low. However, the estimated true seroprevalence was approximately five times higher than the reported SARS-CoV-2 infection based on a molecular test result.

Phylogeography of Human and Animal Coxiella burnetii Strains: Genetic Fingerprinting of Q Fever in Belgium

Q fever is a zoonotic disease caused by the bacteria Coxiella burnetii. Domestic ruminants are the primary source for human infection, and the identification of likely contamination routes from the reservoir animals the critical point to implement control programs. This study shows that Q fever is detected in Belgium in abortion of cattle, goat and sheep at a different degree of apparent prevalence (1.93%, 9.19%, and 5.50%, respectively). In addition, and for the first time, it is detected in abortion of alpaca (Vicugna pacos), raising questions on the role of these animals as reservoirs. To determine the relationship between animal and human strains, Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) (n=146), Single-Nucleotide Polymorphism (SNP) (n=92) and Whole Genome Sequencing (WGS) (n=4) methods were used to characterize samples/strains during 2009-2019. Three MLVA clusters (A, B, C) subdivided in 23 subclusters (A1-A12, B1-B8, C1-C3) and 3 SNP types (SNP1, SNP2, SNP6) were identified. The SNP2 type/MLVA cluster A was the most abundant and dispersed genotype over the entire territory, but it seemed not responsible for human cases, as it was only present in animal samples. The SNP1/MLVA B and SNP6/MLVA C clusters were mostly found in small ruminant and human samples, with the rare possibility of spillovers in cattle. SNP1/MLVA B cluster was present in all Belgian areas, while the SNP6/MLVA C cluster appeared more concentrated in the Western provinces. A broad analysis of European MLVA profiles confirmed the host-species distribution described for Belgian samples. In silico genotyping (WGS) further identified the spacer types and the genomic groups of C. burnetii Belgian strains: cattle and goat SNP2/MLVA A isolates belonged to ST61 and genomic group III, while the goat SNP1/MLVA B strain was classified as ST33 and genomic group II. In conclusion, Q fever is widespread in all Belgian domestic ruminants and in alpaca. We determined that the public health risk in Belgium is likely linked to specific genomic groups (SNP1/MLVA B and SNP6/MLVA C) mostly found in small ruminant strains. Considering the concordance between Belgian and European results, these considerations could be extended to other European countries.