Transboundary and Emerging Diseases

In recent years, significant changes have been observed in the distribution and abundance of local Dermacentor reticulatus populations. However, changes in D. reticulatus dynamics have not been studied in southeastern Poland. Our objective was to enhance our understanding of the environmental factors influencing the occurrence and density of D. reticulatus in this area. Additionally, we sought to investigate the genetic diversity of the tick population and the prevalence of tick-borne pathogens (TBPs). To this end, we established 45 study sites in the Subcarpathian province. Ticks were collected during their peak activity in both spring and autumn. A subset of randomly selected specimens underwent molecular analysis for TBPs screening, using high-throughput microfluidic real-time PCR. Positive amplicons were then sequenced, and phylogenetic analyses were conducted. Our findings confirmed the presence of D. reticulatus ticks in 24 surveyed sites, primarily concentrated in the northern and eastern parts of the region. The mean density of D. reticulatus ticks in their compact range was 5.8 ± 6.4 specimens/100 m². Notably, air temperature and altitude emerged as significant factors influencing the species’ activity. We also identified a high prevalence of Rickettsia raoultii infections in adult D. reticulatus, reaching up to 84.21%. Additionally, 9.52% of ticks were found to be infected with R. helvetica and 4.76% with Anaplasma phagocytophilum. Furthermore, our genetic analyses confirmed the identity of D. reticulatus in the Subcarpathian region, aligning with haplotypes found in other regions of Poland, Czechia, Croatia, and Portugal. In conclusion, our study suggests that the surveyed region represents the current boundary of the compact range of D. reticulatus in Poland in which this tick species exhibits low genetic diversity and a narrow spectrum of detected TBPs.

Elizabethkingia miricola is an emerging nosocomial pathogen responsible for meningitis, sepsis, urinary tract infection, pneumonia, and joint infection in humans. These pathogens were also reported to be causal agents for meningitis-like disease in cultured frogs, which displayed high infectivity, mortality, and significant loss. In July 2023, 10 outbreaks of infectious meningitis-like disease in bullfrogs occurred in Tangshan area. To determine the causal agent, 70 diseased frogs from 10 farms were collected for etiological identification. Gram-negative bacilli were isolated from the brain and liver of sick bullfrogs and identified as members of E. miricola by biochemical characterization and 16S rRNA sequencing analysis. A total of 42 strains of E. miricola were isolated and further determined as the etiological agent by reproducing neurological symptoms and deaths in an artificial infection test. A representative isolate, HBTS-1, was picked up for the pathogenicity test, and the data showed that this stain was highly pathogenic to bullfrogs with an LD₅₀ of 3.7 × 10⁵ CFU. Notably, the isolate also showed high pathogenicity to 5-day-old suckling mice, with an LD₅₀ of 3.1 × 10⁶ CFU, indicating its potential threat to mammals. Moreover, all the 42 E. miricola isolates showed resistance to multiple antibotics without an apparent inhibition zone observed in the test, making the choice of antimicrobial therapy challenging. These novel findings prioritized E. miricola as an important zoonotic agent, which may provide a reference for human medicine.

Canine coronavirus (CCoV) is a common agent of gastroenteritis in dogs, although some variants have been found associated with systemic and often fatal diseases. Distinct genotypes (CCoV-I and CCoV-II) and subgenotypes (CCoV-IIa and CCoV-IIb) are worldwide distributed. In Italy, CCoV infections have been occasionally evaluated, but information about the molecular epidemiology and the genomic features of currently circulating strains is limited. This study reports the detection and molecular characterization of CCoV strains from samples collected from 284 dogs in Italy between 2019 and 2021. CCoV RNA was detected in 39 (13.7%) dogs, as a single viral agent (5 animals, 12.8%) or with other viral pathogens (canine parvovirus types 2a/2b/2c; canine adenovirus type 1; norovirus GIV.2) (34 animals, 87.2%). A total of 48 CCoV strains were detected either alone (CCoV-I: 51.3%, CCoV-IIa: 20.5%) or in copresence (CCoV-I and CCoV-IIa, 23.1%); surprisingly, CCoV-IIb was not identified in this study. Five clusters of CCoV-I were detected, and their spike gene sequences showed the highest nucleotide identities with CCoV-I strains collected from Greece in 2008/2009 and from China in 2021. CCoV-IIa spike gene sequences (three variants) had the highest nucleotide identities with CCoV-IIa strains collected in Greece in 2008/2009 and in Italy in 2009/2011. Given the high CCoV diversity and the variable pathogenicity potential, we underline the need of further surveillance studies to increase our understanding of the epidemiology and evolution of these viruses.

Farm biosecurity has gained increasing attention worldwide during the last decades because of its role in reducing the occurrence of diseases and improving animal performance. Recently, recommendations to reinforce the concept of farm biosecurity in lower- and middle-income countries have been advised. Therefore, this review aims to provide a comprehensive description of the methods and tools used to assess biosecurity compliance in livestock farms in Africa and formulate recommendations. The present review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis extension for scoping reviews guidelines. Peer-reviewed studies reporting biosecurity assessment in poultry, cattle, pig, goat, or sheep farms in Africa were included. Five databases were searched with no date restrictions. A total of 41 studies across 17 countries were finally selected. Selected studies were all published after 2008, and an increasing trend in the number of papers published per year was noticed. In total, 41 different methods for biosecurity assessment were found to be used in African countries, meaning that even within the same country, the same animal species, and the same farming system, different methods were utilized. In many papers, the methods used for biosecurity evaluation were poorly described. In addition, during the biosecurity assessment, measures related to the purchase of laying hens, egg transport and management, calves, calving and dairy management, and nursery units were almost not considered. These measures should be contemplated in future studies since they are related to important risk factors for the introduction and dissemination of infectious diseases. Interestingly, some measures not considered in European biosecurity tools were identified in the selected studies. The observed high difference in methods used may be due to the lack of regulations on biosecurity in African countries; therefore, the authors recommend the development and implementation of a harmonized and contextualized method for the assessment of biosecurity in livestock farms in Africa.

Parrots are traded globally and pose a substantial risk for disease transmission involving parrot-specific pathogens. Parrot bornavirus (PaBV) belongs to the Bornaviridae family and encompasses two clades: alphapsittaciforme (PaBV-1 to -4, PaBV-7, and -8) and betapsittaciforme (PaBV-5 and PaBV-6). These clades cause proventricular dilatation disease, a chronic disease affecting all parrot species. PaBV infections can persist for varying durations in parrots, but the transmission routes are still not well understood. Therefore, surveillance of PaBV-infected parrots is necessary for disease control and improving psittacine aviculture. This study used isolated PaBV-4 NTUCL7 and PaBV-5 NTUCL54 strains to establish and validate two serological diagnostic methods: immunoblotting (IB) and immunocytochemical staining (ICC). To determine the prevalence of PaBV in parrots in Taiwan, 370 clinical serum samples were collected from 13 collaborative veterinary hospitals during a 1-year surveillance period. Serological surveillance revealed a seropositivity rate of 25.68%. Among the seropositive samples, 91.58% were infected with alphapsittaciforme PaBV, demonstrating the predominance of this viral clade in parrots. An analysis of risk factors also demonstrated an association between seropositivity and parrot genera, age, and clinical signs. Cohen’s kappa coefficient analysis showed a high degree of similarity (kappa value = 0.975) between the IB and ICC results, which shows that these serological diagnostic measures are robust. This study established two reliable serological diagnostic measures that are instrumental in serological surveillance, particularly in one of the major parrot-exporting regions. The surveillance results increase the understanding of PaBV infection and associated risk factors and allow methods to be devised for the conservation and protection of parrot populations.

Wild boar (Sus scrofa) is a common wild ungulate known as the most important reservoir of tuberculosis (TB) in Spain. The severity of TB lesions in this species and the high prevalence of porcine circovirus type 2 (PCV-2) have been related. PCV-2 is ubiquitous in swine populations, being usual for the free-living ones the contact with this agent. Recent studies found a correlation between a decrease of generalised TB prevalence in wild boar populations and the PCV-2-vaccination. The aim of this study was to find out if PCV-2 vaccination modulates the gene expression of cytokines from immune cells after its exposition with mycobacterial antigens using an in vitro methodology. A total of 46 wild boars from a PCV-2 infection endemic area were blood-sampled before and after the PCV-2 vaccination of 22 of them. Peripheral blood mononuclear cells (PBMC) were obtained and isolated from these samples. Aliquots of the cells were in vitro cultured and respectively stimulated with PPDa, PPDb, and a mitogen. A complete analysis of the gene expression of cytokines from the cultured PBMC was carried out. Also, Mycobacterium bovis and PCV-2 contacts were revealed by ELISA and/or qPCR. The results demonstrated that the animals which have had contact with PCV-2 and had been vaccinated, manifested a significant decrease in gene expression of proinflammatory cytokines, like interleukin 1 beta, interleukin 6, and tumour necrosis factor-alpha, possibly related with the severity of TB lesions, and also a significant decrease of interleukin 10, a key cytokine. In conclusion, in case of possible infection or contact events with the virus, PCV-2 vaccination could be an effective measure to reduce the TB severity in wild boar populations, which could decrease the intra and interspecies transmission of TB.