Veterinaria Italiana (Journal)
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R07.3 Forecasting West Nile Virus Circulation through Sentinel and Landsat Imagery and Graph Neural Networks
No full textThe West Nile Virus (WNV) infection represents one of the most common mosquito-borne viral zoonosis. Its circulation is highly influenced by climatic and environmental variables influencing vector proliferation and virus replication (Paz & Semenza, 2013). Several statistical models have been developed to predict WNV circulation. Beyond this, the recent massive availability of Earth Observation data, coupled with the continuous advances in the field of Artificial Intelligence, offers new valuable opportunities in modelling animal diseases or vector population dynamics (Porrello et al., 2019).
In the work presented herein, we seek to forecast WNV circulation by supplying Deep Neural Networks (DNN) with high-resolution multi-band Copernicus Sentinel-2 and Landsat-8 imagery. While aggregated indices (e.g. NDVI) have been extensively shown to hold environmental and climatic features, here we let the DNN learn directly from raw spectral bands.
While previous DNN approaches analyze each geographical site independently (Porrello et al., 2019), we propose a spatial-aware approach that includes the characteristics of close sites. Specifically, we build upon Graph Neural Networks to aggregate features from neighbouring locations, while considering their mutual environmental relations. In particular, for each site, we measure their difference in Land Surface Temperature and Surface Soil Moisture - acquired from Copernicus Sentinel-3 and Sentinel-1 respectively - as well as their geographical (Haversine) distance. Moreover, we take into account the temporal factors that characterize the spread of the virus by injecting time-related information directly into the model.
To prove the merits of our proposal, we design an experimental setting that combines satellite imagery with “on-the-ground observations”, i.e. the veterinary cases detected in Italy in the years 2017-2019 and reported in the National Disease Notification System of the Italian Ministry of Health, as described in Candeloro et al., 2020.
By means of both qualitative and quantitative results, we show that our proposed Multi-Adjacency Graph Attention Network (MAGAT) consistently leads to higher performance than classical Machine Learning and Deep Learning baselines, especially when paired with an appropriate pre-training stage
Seroprevalence of Newcastle disease virus and avian influenza virus in poultry and captive wild birds in poultry-dense regions of Pakistan
Get PDFNewcastle disease virus (NDV) and avian influenza virus (AIV) are causing contagious diseases in chickens and wild birds worldwide; however, there is a paucity of information on the current status of seropositivity of Newcastle and avian influenza diseases in chickens and wild birds of Pakistan. Therefore, the present study aimed to investigate the serological evidence of both diseases in commercial poultry (broiler, layer chickens), backyard poultry, and captive wild birds in poultry‑dense regions of Punjab, Pakistan. Enzyme‑linked immunosorbent (ELISA) and haemagglutination inhibition (HI) assays were performed for the determination of antibodies against NDV and AIV and their genotyping and subtyping, respectively. Overall, 47.5% and 67.4% seroprevalence of NDV and AIV, respectively, was observed in both poultry and wild birds. Based on bird’s category, layer chickens had the highest seroprevalence of NDV (60.8%, 95% CI: 52.95‑68.22, OR: 0.71) followed by backyard poultry (56.8%, 95% CI: 47.92‑65.32, OR: 0.82), broilers (52.7%, 95% CI: 46.84‑58.64), pigeons (41.3%, 95% CI: 30.53‑52.81, OR: 1.59), peafowls (26.1%, 95% CI: 11.09‑48.69, OR: 3.16), ducks (23.8%, 95% CI: 12.59‑39.8, OR: 3.57), turkeys (16.7%, 95% CI: 4.41‑42.27, OR: 5.58), parrots (14.3%, 95% CI: 2.52‑43.85, OR: 6.70) and quails (2.3%, 95% CI: 0.2‑13.51, OR: 4.8). Comparatively, backyard chickens had the highest seroprevalence of AIV (78.8%, 95% CI: 70.64‑85.22, OR: 0.63) followed by ducks (73.8%, 95% CI: 57.68‑85.6, OR: 0.83), layers (73.5%, 95% CI: 65.98‑79.89, OR: 0.84), pigeons (72.5%, 95% CI: 61.2‑81.61, OR: 0.89), broilers (70.1%, 95% CI: 64.44‑75.29), turkeys (55.5%, 95% CI: 31.35‑77.6, OR: 1.87), peafowls (47.8%, 95% CI: 27.42‑68.9, OR: 2.56) and parrots (42.8%, 95% CI: 18.8‑70.3, OR: 3.1). Overall, 40.1%, 34.2%, 31.3%, and 25.1% sera were positive for H9 AIV, G‑VII NDV, H7 AIV, and G‑VI NDV, respectively. The current study revealed a widespread exposure to NDV and AIV in poultry and captive wild birds. Therefore, it is crucial to include captive wild birds in NDV and AIV surveillance programs to further strengthen disease control measures, particularly in endemic regions
The occurrence and resistance of Shigella flexneri CECT4804 to acid stress in vitro and in vivo
No full textThe ability to maintain intra‑cellular pH is crucial for many microbes mainly the enterobacteria to survive in diverse environments, particularly those that undergo fluctuations in pH. In this context, the growth and survival of Shigella flexneri at different acid pH values were evaluated to explain the response strategies involved in the adaptation of S. flexneri CECT4804 in acid stress conditions. Furthermore, the capacity of this strain to produce slime on Congo Red Agar, biofilm formation on polystyrene plate and hydrophobicity are reported. In addition, the modification of the membrane fatty acids profiles has been studied. Moreover, an infection with the stressed strain was realized on rats in rates and examined for their toxicity in intestine tissue. The obtained results show that the strain survival is strongly influenced by acidity. Indeed, the stressed and unstressed strains became biofilm positive after acid stress. A significant increase in the hydrophobicity percentage compared to the values found under normal conditions, is also noticed. For the membrane fatty acids analysis, the acidity induces several modifications in the membrane composition. After the infection, the gravest lesion was registered in the intestine of rats administered with the bacteria stressed at the lowest pH
Cross-sectional and histopathological studies of Feline Coronavirus infections in stray cats in Kuwait
Get PDFFeline Coronavirus (FCoV) is a worldwide viral infection of felids. The disease is usually asymptomatic, but it can cause mild diarrhoea; however, few numbers of cases may develop a severe systemic disease known as feline infectious peritonitis (FIP). This study aims to determine the prevalence of FCoV shedding in the faeces of stray cats in Kuwait and detect antibodies against FCoV in their serum. Histopathological analyses and RT‑PCR were used to prove cases of FIP. A total of 178 cats were examined for the presence of FCoV in their faeces using a rapid immunochromatography (IC) test. Anti‑FCoV Antibody (Anti‑FCoV Ab) was detected in their serum using ELISA. Eleven samples were tested using RT‑PCR to confirm positive cases. The prevalence of FCoV faecal antigen in stray cats was 32.6%. The overall detection rate of Anti‑FCoV Ab in stray cats was 44.9%. Nine cats tested positive using the RT‑PCR test. Six out of those nine were confirmed to be FIP positive through gross and histopathological examination. The characteristic uveitis and discoloration of the irises were seen. The present study is the first report confirming FCoV infection in stray cats in Kuwait. Postmortem and histopathological lesions in cases of FIP were recorded
Simulated African Swine Fever (ASF) virus detection in Italy: average numbers of farms and pigs under restriction
Get PDFAfrican swine fever is a devastating contagious viral disease of kept and wild porcine animals that will challenge the Veterinary Services involved in its eradication. Nowadays, ASF represents one of the biggest challenges for the pig sector at a global level. Following a number of simulated virus random introductions, the paper estimates the average number of farms (including their type) and animals that will be under restriction, and finally the average distance of infected farms from the nearest rendering plant. The study includes data referring to 101,032 farms with 9,322,819 pigs which are available in the Italian National Database (BDN). The simulations consider 5 different biogeographic regions with their own domestic pig distribution, breeding systems, and wild boar presence. Following an index case in a farm, and in the worst‑case scenario, in the 10 km radius of the restriction area, there will be: 2,636 farms in South Italy; 470,216 animals in Po Valley; 147 km in Central Italy is the longest mean distance from the infected farm to the nearest rendering plant
O4-5 TIRAP exon 5 polymorphisms influence the outcome of B. melitensis infection in goats
No full textTIRAP is an adaptor protein necessary for TLR4/2 inflammatory signaling in response to bacterial infection. The Brucella effector protein TcpB induces TIRAP degradation, down-regulating the immune response and enhancing Brucella intracellular replication. Two single nucleotide polymorphisms (SNPs, S180L y D96N) in TIRAP exon 5 (TIRAP-E5) were associated with an enhanced resistance to M. tuberculosis, P. falciparum and other infections in humans. The aim of this work was to evaluate if SNPs within caprine TIRAP-E5 influencethe susceptibility to B. melitensis infection in goats. A candidate-gene association study was conducted using 100 DNA samples from Brucella-seropositive (cases) and seronegative (controls) female creole goats. Animals were unvaccinated against Brucella spp. and belonged to ten flocks with prevalence of brucellosis (10 -72%). Genotyping of TIRAP-E5 was carried out by PCR and DNA sequencing. Genetic associations were evaluated by the Fisher’s exact test. Monocyte derived macrophages (MDMs) cultures were obtained from peripheral blood of seronegative goats, and infected with B. melitensis 16 M (MOI 1:10) for 30 min. At different times post-infection, intracellular Brucella counts and cytokine mRNA expression were determined by a gentamicine protection assay and qRT-PCR after RNA isolation, respectively. Sequence alignment of caprine and human TIRAP-E5 showed an identity of 76%. We detected three polymorphic SNPs (rs914(T/C), rs459(C/G) and S189N(G/A)) at TIRAP-E5 in the studied goat population. The rs914-C allele was significantly more frequent in controls than cases (p=0.002). Moreover, the genotypes rs914-CC+CT and the haplotype C-C-G (rs914- rs459-S189N) were associated with absence of Brucella-specific antibodies (p=0.001), while rs459-CG and T-C-A were associated with presence of Brucella-specific antibodies (p=0.043; p=0.045). Bioinformatic analyzes suggested that the associated polymorphisms could affect RNA folding and/or protein function. Besides, MDMs with the rs914-CT genotype controlled more efficiently Brucella intracellular growth at 24 h post-infection (p.i.) (p=0.02), and expressed higher levels of pro-inflammatory cytokines IL-12 e IL-18 at 4 h p.i. than rs914-TT MDMs (p < 0.05). Consistent with previous reports in humans, the results presented here suggest that genetic variability in TIRAP exon 5 would influence the outcome oBf.melitensis exposure in goats
O1-6 Atypical Brucella spp. are facultatively anaerobic in the presence of nitrates, correlating with increased expression of denitrification genes under anoxic conditions
No full textRespiration is a fundamental process in living cells, resulting in ATP production following electron transfer from low-redox- potential electron donors to high-redox-potential electron acceptors such as O2. In prokaryotes, enhanced respiratory flexibility allows the use of alternative electron acceptors, including nitrogen oxides, which contributes to their ability to colonize microaerobic or anaerobic environments. So far, Brucella spp. have been classified as strictly aerobic bacteria, defining O2 as the sole electron acceptor. Use of nitrate as alternative electron acceptor has been suggested, as all genes involved in the denitrification pathway are present in Brucella, but data on possible growth under anaerobic conditions in the presence of nitrates have not yet been available. Comparative RNA-Seq analysis of zoonotic Brucella suis 1330 and the atypical species B. microti grown to mid-log in screw-cap tubes containing Tryptic Soy broth revealed specific upregulation of nor and nos denitrification genes in B. microti. Under anoxic conditions, both species grew in broth, but in the presence of NaNO3 only, and in a NO3-concentration-dependent manner for B. microti. Growth of B. suis was limited, as compared to classical conditions, whereas B. microti, as well as other atypical species such as B. inopinata BO1 and strains isolated from Australian rodents and African bullfrogs, were well-adapted to anaerobic conditions in the presence of NaNO3 as electron acceptor. narG mutants of B. microti and B. suis, lacking nitrate reductase, lost growth capacities under anoxic conditions. Remarkably, denitrification kinetics in the absence of oxygen revealed significantly earlier and faster nitrite production and consumption in B. microti, correlating with RT-qPCR-monitored higher expression of nir, nor, nos genes encoding the corresponding reductases of the denitrification pathway. To our knowledge, this is the first experimental evidence of NO3-dependent anoxic growth of Brucella spp., showing that, in contrast to classical Brucellae, atypical species and strains grow equally well under oxic and anoxic conditions. At least the novel Brucella spp. should therefore definitely be considered as facultative anaerobic bacteria
P7-01 Brucellosis of Veterinary Health Care Workers Engaged in Brucella Vaccination Program in West Bengal, India
No full textRecently the National Animal Disease Control Program in India targeted a massive Brucella vaccination program. In the state of West Bengal about 6000 Veterinary healthcare assistants participated in the program in the month of September, 2021. Among them more than 400 Veterinary healthcare assistants developed symptoms and signs suggestive of brucellosis. Laboratory tests based on STAT, RBT, and ELISA indicated 13% confirmed Brucellosis cases. However, there were also many asymptomatic cases which were not tested. Most of them stated history of needle prick injury during the vaccination program, which was mainly due to disturbed animals. Manifestations started after about a month of the prick injury. Few of them also complained regarding their improper training and supply of proper personal protective equipments. All of them treated in referral hospitals and a standard protocol of Government Health Department was followed. During follow up, it was found that some of them showed gross psychiatric problems. Thus proper training of the Veterinary healthcare assistants is necessary and care should be taken during vaccination of animals. A prophylactic treatment in healthcare assistants with needle prick injuries should be given, and supply of personal protective equipments should be monitored
P1-03 A case report on a novel, non-core Brucella strain isolated from a White´s Tree frog in Germany
No full textIn the past 15 years many divers strains have been assigned to the genus Brucella that differ from the homogenous classical core Brucella species. These novel, atypical Brucella spp. are gaining increasing interest in the scientific community while major information about their epidemiology, hosts, reservoirs, pathogenicity, and zoonotic potential are still missing, and more and more isolates are being reported. Inour case report, we describe brucellosis in a pair of White’s Tree frogs (Litoria caerulea) kept in a German zoo. Differential phenotyping of bacterial cultures was done with classical microbiological methods. Illumina sequencing reads were used for genomic characterization and phylogenetic positioning of the Brucella strain within the genus. The seriously ill White’sTree frogs were submitted for pathologic examination to the responsible chemical and veterinary investigation office. Both frogs were in a poor nutritional status and were presented with additional abnormalities, e.g. skin ulcerations and edema. Bacterial cultures from several organs of both frogs were positive for Brucella and were confirmed by polymerase chain reaction. A Brucella sp. isolate was forwarded to the German Federal Institute for Risk Assessment for in-depth analysis. This isolate displayed non-fastidious and rapid growth and predominantly showed microbiological characteristics similar to other novel, atypical Brucella strains, such as Brucella inopinata-like BO2 or Brucella sp. UK8/14, the latter also isolated from a White’sTree frog. Genomic and phylogenetic characterization placed the isolate apart from the core clade within the novel, non-core clade together with various amphibian and human isolates (inopinata- clade). In summary, we here report another case of amphibian brucellosis caused by anovel, atypical and non-core Brucella strain that clusters together with and is alike to previously reported isolates from amphibian hosts and human brucellosis patients underlining that exotic frogs are potential reservoirs for Brucella spp., posing an underestimated zoonotic hazard for exposed individuals (e.g. exoticanimal keepers, veterinarians)
O6-4 Swab types and storage conditions affect Brucella recovery and DNA detection
No full textBrucellosis is a worldwide zoonosis that once established, is very difficult to eradicate with persistence in animals, environment and humans. The classical Brucella species, B. abortus, B. melitensis and B. suis affect mainly cattle, small ruminants, and pigs, respectively, causing abortions and infertility. Detection of bacteria in live animal can be very challenging, since infected animals do not show systematically clinical symptoms and excretion of bacteria in bodily fluids can be rare and/or intermitted. Sampling with swabs (genitalia, tissue lesions and contaminated environments) from seropositive animals with or without clinical symptoms, can be very useful for bacterial detection and isolation in bacteriological and/or molecular diagnostics. However, no comparison of swab types has been performed in order to ensure successful detection of Brucella. The aim of this study was to compare main commercially used types of swabs for sampling and diagnostics of Brucella spp., and determine the optimal storage conditions for testing. To achieve this, we tested bacterial and molecular methods for detection of classical Brucella species using nine swab types, all with different tip materials, treated immediately after spiking, after 72 h at +4°C, and after 72 h at -20°C. Our results show how storage conditions, freezing (-20°C) vs refrigeration (+4°C), negatively affect Brucella survival even on swabs compared to direct treatment, while no impact on qPCR detection was observed. We demonstrate that Brucella recovery and detection capacity strictly depend on swab tips type, independent of the storage conditions. In particular, bacterial survival is highly variable regarding the type of swabs used for isolation and storage conditions, while molecular detection is uniform. Interestingly, a new type of flocked swab, which includes a conservation medium, showed the highest levels of recovery in both bacterial culture and qPCR, irrespective of the storage conditions (20% higher with than without protective medium). The use of this swab provide increased Brucella viability at refrigeration and freezing temperatures, which significantly increases the diagnostic sensitivity compared to the other tested swabs. To further evaluate performances of flocked swabs under field condition, a multi-centric study targeting infected animals will be conducted within the European national Laboratory Network