Veterinaria Italiana (Journal)
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The immunogenicity and efficacy of acommercially available Infectious Bovine Rhinotracheitis (IBR) virus vaccine against a Pakistani field IBR strain
Get PDFInfectious bovine rhinotracheitis (IBR) is a highly communicable disease of cattle and wild ruminants that is caused by Bovine alphaherpesvirus 1 (BoHV‑1). For IBR control, several developed countries have adopted the immunization and eradication programs focusing on IBR‑positive animals. In Pakistan, livestock producers are importing commercially available vaccine of BoHV‑1, but no studies on the efficacy of these commercial vaccines against local isolates are available. Therefore, the present study was aimed to evaluate the efficacy of a commercially available vaccine of BoHV‑1 against local field isolates of virus. The rabbit model was used and the vaccine was evaluated for immunogenicity and protection after challenge with a highly virulent strain of a field virus. The immune response was measured by virus neutralization titers (VNT). This vaccine induced a humoral response in rabbits but that was not sufficient to completely protect the vaccinated animals against the wild‑type BoHV‑1 strain challenge. While a low virus titer compared to control rabbits was observed in the vaccinated rabbits (p<0.05), there was no sterilizing immunity or freedom from infection. However, complete freedom from disease, for example, the absence of pyrexia was noticed in the vaccinated group. In conclusion, the present study demonstrated that imported vaccine stock provoked only a partial protection against indigenous isolated of BoHV‑1. However, tests performed on rabbits are preliminary, as only those performed on the source species can determine more reliable results
Infezione da Strongyloides stercoralis e follow-up a lungo termine in un cane di proprietà nel nord ovest dell'Italia
No full textStrongyloides stercoralis is a zoonotic parasite, infecting human, cats and dogs. The infection is typical of tropical and subtropical areas, while it is underestimated in temperate countries. In Europe, data from humans demonstrate a large variability of infection prevalence; while scarce literature is available for dogs, with most cases reported in kennel dogs from southern Italy. A privately owned 8-month-old French Bulldog was examined because of chronic cough, diarrhea, and poor body conditions. Radiography of the thorax revealed a diffuse bronchial pattern and abdominal ultrasound showed unspecific signs of inflammation. Cytological analysis of a rectal swab revealed the presence of larvae compatible with Strongyloides sp. A parasitic infection causing protein-losing enteropathy and bronchopneumonia was suspected and fenbendazole was administered with a poor clinical response. Fecal Baermann test revealed S. stercoralis rhabditiform larvae and thinshelled eggs. Clinical signs resolved progressively after two subcutaneous ivermectin administrations (off-label) given two weeks apart. Serial coprological controls scheduled in the following 6 months were negative for larvae and eggs. Fecal real-time PCR confirmed negative results 6 months after treatment. The authors describe for the first time a case of S. stercoralis infection in a privately-owned dog from north-west of Italy (Piedmont)
Serological Investigation of Japanese Encephalitis Virus Infection in Commercially Reared Pigs, Southwestern Nigeria
Get PDFJapanese encephalitis virus (JEV) is a zoonotic arbovirus that causes abortion, stillbirth, and congenital defects in pigs, and epidemic encephalitis in humans. Currently, there is scarcity of information on JEV infection in pigs in Nigeria. Since the Culex tritaeniorhynchus vector of JEV is present in Nigeria and considering recent anecdotal reports of abortions and birth of weak piglets in some pig farms in southwestern Nigeria, there is a need for studies on the presence of the virus and its true burden among pig populations in the country. Serum samples (n=368) obtained from farm-reared pigs in four States of southwestern Nigeria were screened for JEV-specific IgG antibodies using a commercial ELISA kit. An overall JEV seropositivity of 35.1% (95% CI: 30.18 – 39.93%) was obtained, with detectable antibodies in pigs of all age groups, breeds, sex, and locations. Our results suggest natural exposure of these unvaccinated intensively reared pigs to JEV circulating silently in the swine population with significant association of the seropositivity with location (state/community in which the pig farms exist) and breed of the pigs studied. This first report of detection of anti-JEV antibodies in pigs in Nigeria indicates that JEV circulated among these pigs and underscores the need for active surveillance for JEV in humans, pigs, and mosquitoes to provide valuable epidemiological data for the design of effective control strategies against the virus, thus forestalling potential future outbreaks of the infection
Leishmania infantum surveillence on dogs from Primavera do Leste, Mato Grosso state, Brazil.
Get PDFThe objective of this research was to determine the seroprevalence of Leishmania infantum in dogs in the city of Primavera do Leste, Mato Grosso, Brazil. A total of 109 serological samples from dogs were analyzed, collected between August 2021 to March 2022, using rapid immunochromatographic test, TR DPP® (LVC BioManguinhos), and Immunoenzymatic Assay (ELISA), EIE-Canine Visceral Leishmaniasis (Bio-Manguinhos®, Fiocruz). IgG anti-Leishmania infantum antibodies were detected 4.6% (5/109) by DPP test. Regardless of results, all samples were submitted to ELISA test, of those 0.9% (1/109) was positive, the same having also been positive for the DPP, from asymptomatic, adult dog living in the urban area. This is the first autochthonous report of CVL in the municipality. Since Visceral leishmaniasis caused by L. infantum is a disease of One Health concern, the results obtained are essential to fill the epidemiological gap of in the region. Furthermore, the health authorities should implement surveillance actions in order to monitoring of dogs, as well as survey of sand flies, and health education programs to stimulate the prevention of the disease
Culicoides species responsible for the transmission of Epizootic Haemorrhagic Disease virus (EHDV) serotype 8 in Italy
Get PDFEpizootic haemorrhagic disease (EHD) is a viral disease transmitted by Culicoides biting midges that affects wild and domestic ruminants. The causative agent, EHD virus (EHDV), belongs to the family Sedoreoviridae, genus Orbivirus. The virus has never been reported in Europe until October 2022, when the virus was for the first time detected in Sicily and Sardinia. After the first clinical cases, an intensive entomological field activity was carried out in five EHD affected farms located in Sardinia, with the aim of assessing the EHDV vector competence in European species of Culicoides. EHDV‑8 was detected in C. imicola, C. obsoletus/scoticus, C. newsteadi, C. pulicaris ss, and C. bysta. The first 4 species have also been demonstrated to be able to transmit bluetongue virus (BTV). According to these results, it is likely that EHDV‑8, sharing the same transmission patterns of BTV, can also spread to Europe
P3-01 Manipulating Brucella abortus two-component regulatory system BvrR/BvrS promoter activity through environmental stimuli
No full textThe two-component regulatory system BvrR/BvrS is required for Brucella abortus transition from an extracellular to an intracellular lifestyle. BvrS is a sensor histidine kinase transducing unknown external stimuli to the BvrR transcriptional regulator through phosphorylation. Active BvrR then binds to gene regulatory regions, affecting their transcription. The system is a master regulator controlling the expression of genesrelated to cell envelope homeostasis, carbon and nitrogen metabolism, the virulence factor VirB and its regulator VjbR. B. abortus bvrR/bvrS mutants are avirulent in mice models. Low concentration of nutrients and pH were recently described as environmental cues affecting the BvrR transcriptional regulator activation through phosphorylation. Here, we describe the environmental cues that affect BvrR/BvrS promoteractivity. Using a pbvrR::luxA/luxB transcriptional fusion, the promoter activity was determined during in vitro growth and ex vivo in a cellular infection model. Conditions and chemical compounds simulating the environment found during intracellular trafficking were evaluated: nutrient restrictions, different pH, presence of metals, carbon, and nitrogen sources, at different concentrations and time. The results obtained for each condition tested, individually or in combination will be presented in this report. Some of the assessed conditions affected bvrRp activity and demonstrated a similar effect on BvrR phosphorylation. A combination of conditions tested was found to repress bvrRp activity. Altogether, these results show that conditions that modulate bvrRp activity are not necessarily the same as those sensed by the BvrR/BvrS two-component system but might influence BvrR activation. This work provides a first attempt toin vitro manipulate the bvrRp activity and hence contribute to the understanding of the type of environmental signals need it for regulating bvrR/bvrS transcription
P1-02 Identification of atypical Brucella spp. by MALDI ToF MS
No full textCorrect identification of atypical Brucella spp. and their discrimination against former Ochrobactrum species remains difficult with current routine diagnostic procedures. Matrix-assisted time-of-flight mass spectrometry (MALDI ToF MS) is a fast, reliable, cost-effective and high-throughput technique, increasingly used to identify microorganisms by their proteomic fingerprint. However, successful identification of a specific pathogen is only possible with an accurate reference database that covers the respective genus. Unfortunately, none of the major manufacturers of MALDI ToF MS systems offers databases comprising all currently known Brucella species. To overcome this drawback, various internal pipelines and databases have been established over the last years. Yet, the newly emerging species require an extension of these pipelines due to the lack of reference spectra and biomarkers. We describe the identification of several atypical Brucella strains,including former Ochrobactrum anthropi and Ochrobactrum intermedium, using their MALDI ToF MS spectra by taking advantage of unique, species-specific peak pattern profiles. These specific mass spectral peaks have been identified by comparing spectra of 110 different strains and roughly 2,600 spectra belonging to the six classical and two marine Brucella species, the novel species B. microti, B. inopinata, B. papionis and B. vulpis as well as various atypical Brucella isolates from frogs and rodents. The proteins corresponding to the species-specific peaks were identified using our in-house pipeline combined with data from NCBI. The identification of these species-specific spectral peaks and their subsequent assignment to Brucella biomarkers finally highlights the reliability of our method. In summary, classical and novel Brucella spp. can be quickly and easily identified using the here presented MALDI ToF MS pipeline, avoiding the commonly described misidentifications with former O. anthropi, O. intermedium and atypical Brucella strains. The ability of MALDI ToF MS to discriminate novel atypical Brucella and former Ochrobactrum species from classical Brucella and B. inopinata(-like) is especially important in the context of biological safety regulations
P5-01 Evaluation of different adjuvants associated with vaccination candidate Brucella ovis ∆abcBA in a murine model of infection by Brucella ovis
No full textBrucellosis is an infectious disease caused by facultative intracellular Gram-negative bacteria, of great importance in animal and human health. Among the known species, Brucella ovis is capable of infecting sheep, mainly causing epididymitis and reproductive failure in ram, leading to important economic losses, thus highlighting the need to develop vaccines to help control this disease. Previous studies have shown good levels of protection againstB. ovis infection both in a murine model and in the natural host using the B. ovis ΔabcBA strain abcBA encapsulated by alginate. Therefore, this study aimed to evaluate the increase in the vaccine potential of the candidate B. ovis ΔabcBA associated with different adjuvants, polymeric matrix or alginate microcapsule and Chitosan, in mice challenged with B. ovis. We observed that B. ovis ΔabcBA encapsulated by alginate with chitosan, but not associted to polimeric matrix favored lower bacterial recovery in both the spleen and liver of challenged animals, demonstrating better efficacy in controlling the infection. It should also be noted the ability of this vaccine formulation to lead to a greater synthesis of immunoglobulin production, especially immunoglobulin G, where this increase in total levels is correlated with the increase in certain subclasses, with a significant emphasis on the higher levels of the IgG2a subclass, involved inthe favoring the control of infections by intracellular agents. Addicionally, use of this vaccine formulation resulted in lower score of histopathological lesions in target organs. These results demonstrate the vaccine potential of the B. ovis ΔabcBA strain encapsulated by alginate with chitosan, leading to both a higher protection index, as well as inducing the significant production of antibodies related to a protective immune response, making possible its future evaluation in a natural host
P6-02 Use of Brucella Ovis Antigen for Canine Brucellosis Diagnostics by Agar Gel Immunodiffusion Assay
No full textCanine brucellosis caused by Brucella canis is a zoonotic infectious disease that is common worldwide. This infection is not registered in Ukraine, due to the lack of diagnostic tools. One of the available laboratory diagnostic methods is the agar gel immunodiffusion test (AGID), which is recommended for testing of dogs with suspected brucellosis. The objective of the work was to identify an antigen for AGID to detectantibodies against B. canis. The study was conducted using thermo-extract R- antigen with 4×109 CFU of Brucella ovis 67/B strain. Reference anti-B. canis serum was used as a reference positive sample that was kindly provided by Claire Ponsart (ANSES). Antigen tests were performed by AGID according to George and Carmichael (1978) using field sera (n=547) taken from stray dogs from five regions of Ukraine. For comparison purposes, these samples were also tested using commercial C. Brucella Ab Test Kit / Rapid Test Kit (Bionote, Republic of Korea). The antigen was made by thermo-extraction of Brucella ovis 67/B. The determination of the resulting antigen activity was studied by AGID using the reference anti-B. canis serum in dilutions from 1:5 to 1:40. It was found that the minimum dilution at which a precipitation line was detected in agar is 1:20 in all repetitions. Furthermore, 54 (9.87%) samples of blood serum from dogs were positive, which indicated the circulation of B. canis in stray animals. All AGID positive samples were also confirmed using commercial C. Brucella Ab Test Kit / Rapid Test Kit. Our studies have shown the suitability of the obtained thermo-extracted R- antigen from B. ovis 67/B strain for its use in AGID to detect antibodies against B. canis. We also found a high seroprevalence of canine brucellosis among stray animals in Ukraine that may require the introduction of wide monitoring among domestic dogs
P6-03 The cloning, expression and immunological evaluation of the Omp31 protein from Brucella melitensis and evaluation of its possible use for diagnosis in bovine brucelosis.
No full textThis work describes molecular methodologies for the production of OMP31r for the serological diagnosis of bovine brucellosis, which can be caused by biovars of Brucella abortus. Brucellosis is easily transmitted to man and causes an acute febrile illness, undulating fever, which can become chronic and cause serious multisystem complications. Unequivocal diagnosis of Brucella infection can only be made by isolation and identification of Brucella but in situations where bacteriological analysis is not possible, diagnosis can be based on serological methods. Furthermore, false positives can be expected in vaccinated animals, because the antibodies cross-react with infection by the wild-type strain. Serological monitoring tests typically use total antigen such as: Brucella buffered, i.e. rose bengal test (RBT) or buffered plate agglutinationtest (BPAT), or lipopolysaccharides in various ELISA formats. At this point, selecting one or more antigens and using molecular biology to obtain it in its recombinant form would provide a fast and efficient way to innovate in serological diagnostic methods. The Omp31 antigen has been reported for the diagnosis of brucellosis in sheep and goats. In this work, the Omp31 gene from B. melitensis was isolated to be cloned and expressed in a Escherichia coli, where the protein is obtained safely and in large quantities. In this research work, a wide variety of techniques such as bioinformatics, molecular biology, biochemistry and immunology were applied. Omp31r showed a sensitivity and specificity of 77 and 90% and was not recognized by sera from vaccinated animals and by a hyperimmune serum against B. abortus RB51, which is the strain used as a vaccine, which is important since it does not produce false positives, that is to say, it does not produce crossreaction, and the consequent economic cost for the farmer of slaughtering an animal that is not really sick. The Omp31r has optimal performance and quality for use in practical laboratory work and research projects that require the serological diagnosis of Brucellosis sp. This achievement will undoubtedly allow commercial independence in relation to antigen production, taking a big step at the institutional level on the path towards the production of diagnostic kits against Brucella