Veterinaria Italiana (Journal)
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The Border Disease virus (BDV) prevalence and genetic typing in ruminant flocks in Turkey
No full textThis study aims to update current data regarding Border Disease in sheep and goats, determine the first prevalence of BDV in cattle and identify its circulated genotype in Turkey. For this purpose, 100 sheep, 20 goats and 193 cattle aborted fetuses sent for diagnosis to Samsun Veterinary Control Institute between 2015 and 2017 were analyzed in terms of pestivirus by Ag‑ELISA, BDV by Real‑Time test (RT‑PCR) and Conventional RT‑PCR test. The rate of pestivirus positive animals was found at 50.26% (97/193) in cattle, 58% (58/100) in sheep and 55% (11/20) in goats by the pestivirus Ag‑ELISA test. Total of 58 Ag‑ELISA positive sheep were tested by Real‑Time RT‑PCR and conventional RT‑PCR tests. End of the tests, one sheep sample (1.72%) was found BDV positive by Real‑Time RT‑PCR test and three sheep (5.17%) and one cattle (1.03%) samples were detected as BDV positive by conventional RT‑PCR test. BDV positivity was not detected in goats in this research. All samples that were found positive by conventional RT‑PCR test and Real‑Time RT‑PCR test were genotyped by phylogenetic sequence analysis, and obtained results showed that BDV‑3 and BDV‑7 genotypes of BDV in sheep and BVDV‑1 genotype in cattle circulated in the investigated area. The sequence analysis results revealed that conventional RT‑PCR and Real‑Time RT‑PCR tests detected genotype BDV‑3, while genotype BDV‑7 was only detected by conventional RT‑PCR test in sheep abortion materials. Additionally, it was found that one bovine specimen was BDV positive by conventional PCR, but the same sample was identified as BVDV‑1 at sequence analysis. The obtained data of this study showed that new probes should be designed using our local strains for BDV diagnosis by Real‑Time RT‑PCR assay, and cattle must be sampled for BDV screening, and PCR tests results should always be confirmed by sequence analysis
First record of Aedes japonicus in Liguria region, Northwest of Italy
Get PDFAedes japonicus is an invasive Asian mosquito species, and to date it is widespread in many European countries. In Italy, it was first recorded in 2015 at the Austrian border and it then spread throughout the Northeast of the country. In 2019, it was also identified in Piedmont region, near the Swiss border. In the framework of the Italian program for prevention, surveillance, and response to Arboviruses, from June to November 2021, biweekly entomological surveillance was performed in the Liguria region (Northwest Italy). The collected mosquitoes were morphologically and genetically identified and molecularly analysed for the detection of West Nile and Usutu viruses. Six female mosquitoes, trapped on the 6th of July 2021 using a gravid trap in Albenga (Savona province), were morphologically identified as Ae. japonicus and the identification was genetically confirmed. The pool tested was negative for the presence of West Nile and Usutu viruses. The detection of Ae. japonicus was performed in a coastal area characterized by the presence of many floriculture activities. Considering the distance from the established Ae. japonicus mosquito populations in Italy and other European countries, this could represent an independent introduction in this country
O3-1 BNIP3L-dependent mitophagy induced by Brucella abortus in host cells is required for bacterial egress
No full textBrucella species are Gram-negative facultative intracellular bacteria responsible for brucellosis, a worldwide zoonosis affecting various hosts including humans. Along the infection in most host cells, Brucella first interacts with the endosomal pathway and eventually reaches its replicative niche inside the endoplasmic reticulum (ER), in which bacteria multiply massively. Regarding the many physical and functional interactions between the ER and mitochondria, as well as the major role of mitochondria in inflammation and host-pathogen interactions, we aim to study the effect of Brucella on the mitochondrial population of infected cells and the crosstalk existing between them which is still poorly understood. In this work, we showed that Brucella abortus induces mitophagy through an increase in LC3/mitochondria colocalizing events, as well as an increase in the number of acidified FIS1 mitochondrial fragments at 48h post-infection in host cells. Brucella abortus -induced mitophagy is accompanied by a strong mitochondrial network fragmentation which is dependent on the mitophagy receptor BNIP3L as we demonstrated that a siRNA-mediated silencing of BNIP3L prevents the mitochondrial fragmentation and mitophagy in Brucella abortus infected cells. Our results also show that the expression of BNIP3L induced by Brucella abortus relies on the activation of the hypoxia-inducible factor HIF-1α in an oxygen-independent way. Instead, HIF-1α activation appears to be iron-dependent since FeCl2 supplementation prevents HIF-1α nuclear translocation and BNIP3L expression in Brucella abortus infected cells even if the origin of a putative iron starvation response in the host cells remains to be elucidated. In an attempt to better understand the functional role of the Brucella abortus-in-duced BNIP3L- mediated mitophagy, and what would be the resulting advantage for the host cell and/or the bacteria, our results show that BNIP3L silencing drastically reduced the number of reinfection events at 72 h post-infection, suggesting that bacterial egress from the host cell could be impaired. Future research will be needed to decipher the dynamics and the role of mitophagy and mitochondrial membranes in the last steps of Brucella abortus intracellular cycle. Altogether, those results should highlight new molecular mechanisms and critical steps involved in Brucella trafficking during infection
O3-2 RNA-immunoprecipitation/miRNA-Seq reveals miRNA-like, small noncoding RNAs of Brucella suis, translocated into the cytoplasm of infected murine macrophages
No full textIntravacuolar bacterial pathogens have evolved a variety of strategies to subvert host cell functions during infection. In the past two decades, cellular microbiology has focused on the interplay between bacterial proteins with host cell proteins, lipids and DNA. However, over the same period, small noncoding bacterial RNAs have emerged as central players in gene expression regulation and pathogenesis. Moreover, the translocation of microRNA-like sRNAs (miRNAs) from intracellular bacteria into the host cell cytoplasm to manipulate cellular functions has been recently reported. Using the izMiR miRNA detection algorithm, we bioinformatically identified sequences encoding candidate pre-miRNAs, and which were similar to human pre-miRNAs, in the genomes of several intravacuolar bacterial pathogens. To validate bioinformatics predictions, we immuno-precipitated the RNA-induced silencing complex (RISC) from RAW 264.7 murine macrophages infected or not by Brucella suis 1330. Following RNA extractions, samples from infected and non-infected cells were differentially analyzed by small RNA-Seq. This approach led to the identification of six RISC-associated transcripts specifically aligning with the B. suis genome, matching the izMiR-based predictions, and showing the characteristics expected for authentic miRNA. Among these, B. suis MIR6325 and MIR11484 are of particular interest, since bioinformatics analysis using the MR-microT algorithm led to the identification of highly-ranked candidate eukaryotic targets. Ongoing work is aiming at the experimental validation of host cell target mRNAs for miRNAs MIR6325 and MIR11484, using a RISC-Trap approach. In parallel, we are generating B. suis deletion mutants for several validated pre-miRNAs to evaluate their phenotypes and possible impact in the context of host cell infections by using next-generation sequencing, cellular microbiology, high-resolution microscopy and screening approaches. Our study will shed light on an emerging, yet poorly explored domain of host/pathogen interactions, and may help to identify targets for the development of new biomarkers and antimicrobials
Histopathological and molecular methods as complementary diagnostic in case of lymphadenopathies suggestive of bovine tuberculosis
Get PDFBovine tuberculosis (TB) is a chronically evolving zoonotic infectious disease caused by Mycobacterium bovis. Anatomopathological examination during post mortem inspection in bovines is the main resource engaged in sanitary slaughter; however, it is very troublesome since many granulomatous inflammatory processes have similar morphological characteristics. Thus, this study aims to use complementary diagnosis methods (histopathological and polymerase chain reaction – PCR assays) to confirm the macroscopic assessment of lymphadenopathies indicative of tuberculosis in bovines slaughtered in a refrigerated slaughterhouse in Tailândia city, PA, Brazil. Fifty‑one samples were collected from lesions indicative of tuberculosis in pre‑scapular and pre‑pectoral lymph nodes (or different lymphadenitis) in condemned carcasses. Histological processing employed routine techniques carried out at the Laboratory of Animal Pathology of the Federal Rural University of the Amazon, while the PCR assay was performed at the Bacteriology Laboratory of the Evandro Chagas Institute. Results showed that 1.96% of the histopathology samples corresponded to inflammatory processes typical of TB and that, in PCR, 4.25% of the samples had the amplification profile of the M. bovis species. These results indicate the importance of adding complementary methods to assist the sanitary inspection line and make inspection more efficient in its decisions
INDICE RESISTIVO RENALE NEI GATTI OBESI E NON OBESI
No full textThis study aimed to compare renal function between obese and normal‑weight healthy cats, using intrarenal resistive index (RI), serum symmetric dimethylarginine (SDMA), and serum creatinine, and to identify the variables that might influence intrarenal RI. Thirty crossbred client‑owned cats met the inclusion criteria and were allocated into two groups: Control and Obese. Body weight, body mass index (BMI), body condition score (BCS), SAP, serum SDMA, urea, and creatinine were evaluated. B‑mode and Doppler ultrasound of the kidneys were done. RI evaluation was in the interlobar artery. SDMA and intrarenal RI were compared between groups, also considering the gender of the cats. A correlation analysis between intrarenal RI with the other parameters was performed. SDMA was higher in the Obese group. Intrarenal RI was higher in females than males in the Obese group. Obese females presented higher RI and SDMA than Control females. A positive correlation was observed between RI, age, body weight, and BMI. Six obese cats (40%) showed increased RI. The increase in body weight, BCS, and BMI resulted in a simultaneous increase in RI and SDMA. The RI may assist in monitoring renal function, and may be associated with preclinical kidney changes in obese cats
Pharmacodynamics and pharmacokinetics of nalbuphine in xylazine‑sedated horses
Get PDFThis study describes the selected pharmacodynamics and pharmacokinetics of nalbuphine (NAL) in xylazine (XYL)‑sedated horses. Five adult healthy horses were randomly received 2 treatments at a 1‑week interval; XYL treatment (0.55 mg/kg IV) and XYL/NAL treatment (XYL, 0.55 mg/kg IV; NAL, 0.3 mg/kg IV). The measured pharmacodynamic variables were sedative and analgesic effects and the effect on ataxia and some physiological parameters. for the pharmacokinetics of NAL, its plasma concentrations were measured using HPLC and a 2‑compartment analysis was performed. Greater and prolonged sedation was evident after XYL/NAL treatment compared with XYL treatment. Slightly improved and prolonged analgesia was demonstrated after XYL/NAL treatment. Significant changes in blood pressure and respiratory rate lasted for a shorter duration with XYL/NAL treatment than with XYL treatment. After XYL treatment, rectal temperature was significantly different from baseline and XYL/NAL treatment. Elimination half‑life of NAL was 3.47 ± 1.39 hours and total body clearance was 2.88 ± 0.73 L/kg/hour. In conclusion, addition of NAL to XYL resulted in remarkable advantages on the measured parameters. The obtained pharmacokinetics of NAL could be useful in determining the effective NAL infusion rate, which could be further evaluated as an adjunctive agent to XYL for prolonged sedation in horses
Evidence of West Nile virus in chickens and horses in Nigeria: results from a serosurvey
Get PDFWest Nile virus (WNV) is an emerging arbovirus which affects humans and horses. A cross sectional study was carried out on 106 local horses in Kaduna and 78 domestic chickens in Federal Capital Territory. A total of 184 sera were screened for West Nile virus anti Pr‑E antibodies using ID Screen® West Nile competitive enzyme linked immunosorbent assay. For the horses, an overall prevalence of 92.45% was recorded while domestic chickens had a preponderance of 7.69%. From our study, there was a statistical significant difference between the occurrences of WNV in stallions than mares with p < 0.05. Comparing the occurrence of West Nile virus between species, horses were more likely to be infected by West Nile virus than domestic chickens (OR 147). This is the first seroprevalence study investigating West Nile virus infection in domestic chickens in Nigeria. The presence of the antibodies indicates the widespread circulation and the potential risk of infection in humans and animals. In order to understand the epidemiology of West Nile virus infection in Nigeria, there is need for surveillance to be implemented in human and animal sectors
Evaluation of cardiovascular injury in dogs coinfected with visceral leishmaniasis and monocytic ehrlichiosis by echocardiographic examination and selected biomarker measurements
Get PDFVisceral Leishmaniasis (VL) and Monocytic Ehrlichiosis (ME), which are an important zoonotic diseases of dogs, causing multiple organ dysfunction and has a poor prognosis when not interfered. In this study, it was aimed to investigate the cardiovascular injury that develops in dogs that co‑infected with VL and ME with cardiovascular biomarkers and echocardiographic parameters. The animal material of this study was consisted of 14 owned dogs in total; 7 diseased dogs which were determined to be co‑infected with VL and ME according to the results of clinical examination and rapid test kits, and 7 healthy dogs, which were determined to be healthy as a result of the same examinations. As a result of echocardiographic examinations, decreased left ventricular cytolic and diastolic diameters (LVIDs, LVIDd), fractional shortening (FS) and increased ratio of left atrium to left aortic root diameter (LA/Ao) values were determined in the Co‑infected Group compared with the Healthy Group. Also, as a result of biomarker analysis, higher cTnI) D‑dimer and NT‑proBNP levels were detected in the Co‑infected Group. In conclusion, considering studies of dogs infected with VL and/or ME alone, it was concluded that similar cardiovascular injury develops in dogs co‑infected with VL and ME
O6-5 Proteomics-based identification of immunodominant Brucella canis proteins as candidates for serodiagnosis and vaccine development
No full textCanine brucellosis is an emerging disease and Brucella canis can be a potential health threat for dog holders. Diagnosing canine brucellosis has pitfalls since serological methods that recognize B. abortus and B. melitensis infections fail to detect anti-B. canis antibodies. Therefore, we used an immunoproteomics approach to identify immunodominant proteins as antigen candidates to improve serological testing. We performed two-dimensional gel electrophoresis with B. canis cell extracts followed by MALDI-ToF MS analysis and could correlate 182 protein spots with 82 B. canis proteins. Western Blot analysis of 2D SDS-PAGE gels using the sera from B. canis-infected dogs, verified by culture, PCR or serological tests, and sera from uninfected dogs, detected 50 immunoreactive B. canis proteins. Hence, 32 out of 82 identified proteins did not react with any sera, although seven of these proteins have been identified in previous studies as immunodominant in B. abortus and/or B. melitensis. A total of 14 proteins, like GroEL, DnaK or KatA, were false-positive hits since they reacted with sera of B. canis - free dogs. In contrast, 36 immunogenic proteins were detected by serum from bacteremic dogs and 16 out of these proteins also reacted with serum from infected but non-bacteremic dogs. Some of these immunoreactive proteins, e.g. three periplasmic substrate-binding proteins, seem to be specific for B. canis and have not been described before as immunodominant in Brucella. A subset of immunodominant proteins were also found in B. canis outer membrane vesicles that might serve as vaccination platform. In summary, we identified several immunoreactive proteins for the detection of acute and chronic canine brucellosis