Brucella abortus Strain RB51 Vaccine: Immune Response after Calfhood Vaccination and Field Investigation in Italian Cattle Population

Immune response to Brucella abortus strain RB51 vaccine was measured in cattle vaccinated at calfhood. After an increase at day 6 post-vaccination (pv), the antibody level recorded in the 10 vaccinated animals remained constant for two months, and then progressively decreased. All vaccinated animals remained negative from day 162 pv to the end of the study (day 300 pv). Only at days 13 and 14 pv the RB51-CFT showed 100% sensitivity (credibility interval (CI) 76.2%–100%). The results indicate that the possibility to use RB51-CFT for the identification of cattle vaccinated at calfhood with RB51 is limited in time. A field investigation was carried out on 26,975 sera collected on regional basis from the Italian cattle population. The study outcomes indicate that in case of RB51-CFT positive results observed in officially Brucellosis-free (OBF) areas and, in any case, when an illegal use of RB51 vaccine is suspected, the use of the RB51-CFT alone is not sufficient to identify all the vaccinated animals. The design of a more sophisticated diagnostic protocol including an epidemiological investigation, the use of RB51-CFT, and the use of the skin test with RB51 as antigen is deemed more appropriate for the identification of RB51 vaccinated animals

Brucellosis and One Health: Inherited and Future Challenges

One Health is the collaborative efforts of multiple disciplines to attain optimal health for people, animals and the environment, a concept that historically owes much to the study of brucellosis, including recent political and ethical considerations. Brucellosis One Health actors include Public Health and Veterinary Services, microbiologists, medical and veterinary practitioners and breeders. Brucellosis awareness, and the correct use of diagnostic, epidemiological and prophylactic tools is essential. In brucellosis, One Health implementation faces inherited and new challenges, some aggravated by global warming and the intensification of breeding to meet growing food demands. In endemic scenarios, disease awareness, stakeholder sensitization/engagement and the need to build breeder trust are unresolved issues, all made difficult by the protean characteristics of this zoonosis. Extended infrastructural weaknesses, often accentuated by geography and climate, are critically important. Capacity-building faces misconceptions derived from an uncritical adoption of control/eradication strategies applied in countries with suitable means, and requires additional reference laboratories in endemic areas. Challenges for One Health implementation include the lack of research in species other than cattle and small ruminants, the need for a safer small ruminant vaccine, the need to fill in the infrastructure gap, the need for realistic capacity-building, the creation of reference laboratories in critical areas, and the stepwise implementation of measures not directly transposed from the so-called developed countries.One Healthbrucellosisawarenesscapacity buildingclimateglobal warmingintensificationdiagnosisvaccinesUnpublishe

Canine Leishmaniasis: Serological Results in Private and Kennel Dogs Tested over a Six-Year Period (2009–2014) in Abruzzo and Molise Regions, Italy

This paper reports the results of serological tests for the detection of antibodies against Leishmania spp. in Abruzzo and Molise regions from 2009 to 2014, with the aim of evaluating the presence and distribution of canine leishmaniasis. Data were extracted from the Laboratory Information Management System (LIMS) of the Istituto Zooprofilattico Sperimentale of Abruzzo and Molise, and then the dog identification numbers were matched with those stored in the Canine Registries of the two regions to get information about the age of dogs at time of testing. Dogs were considered positive when having an IFAT (Indirect Fluorescent Antibody Test) titer ≥1:80. In total, 41,631 dogs were tested, 85.3% from Abruzzo and 14.7% from Molise. At the provincial level, the percentage of positive dogs ranged from 5.2% (L’Aquila, Abruzzo region) to 21.8% (Campobasso, Molise region). Findings are consistent with the hypothesis that in the coastal areas, the relationships between the host, the vector, and the agent are more favorable for the spreading of CanL, and it seems that densely populated urban internal areas have less favorable conditions. Being a dog hosted in a kennel seems not to be a factor increasing the probability that dogs show positivity, even in long-term sheltering conditions

Standardisation of an indirect enzyme-linked immunosorbent assay for the detection

An indirect enzyme-linked immunosorbent assay (ELISA) was evaluated for the detection of Brucella antibodies in milk from water buffalo (Bubalus bubalis Linnaeus, 1758). The test accuracy was evaluated on milk samples from the Campania Region in Italy. A total of 100 negative samples were collected from 10 officially brucellosis-free herds in Salerno Province, while 30 positive samples were collected from 3 herds in Caserta Province, where animals gave positive results to the official tests and it was here that Brucella abortus biovar 1 had been isolated. Test sensitivity was 100%, with a confidence interval (CI) of 90.8%-100%, while specificity was 98% (CI 93%-99.4%) on individual milk samples. To simulate bulk milk samples from herds infected at various levels of infection, dilutions from 1:10 to 1:100 of positive milk samples in negative milk were also used. The probability of detecting antibodies in positive milk samples was higher than 50% up to a dilution of 1:50 in negative milk. Considering the average national water buffalo herd size, the probability of identifying infection in a water buffalo herd by bulk milk testing is 50% (CI 33.1%-66.9%) in the worst case scenario of a single infected animal contributing to the bulk milk

An ELISA for the evaluation of gamma interferon production in cattle vaccinated with Brucella abortus strain RB51

The results of an enzyme-linked immunosorbent assay (ELISA) implemented for the detection of gamma interferon (g-interferon) production in cattle vaccinated with Brucella abortus strain RB51 are presented. A purified protein fraction derived from RB51 (RB51 brucellin) has been used as antigenic stimulus for whole blood. The test was evaluated for 300 days in ten heifers vaccinated at calfhood with 10 × 109 colony-forming units of RB51 and in five control heifers. All animals came from officially brucellosis-free herds. Vaccinated animals started to give positive results from day 17 post vaccination (pv) until day 239 pv. All vaccinated animals gave a positive reaction at least once (with a stimulation index exceeding 2.5). Nevertheless, if sampling on day 20 pv is excluded (90% of vaccinated animals gave positive results), the sensitivity of the test varies from 20% to 70%, with a 40% average. A stimulation index over 2.5 was also recorded in three control animals. The results suggest that the g-interferon test is not suitable for the detection of cattle vaccinated with RB51, either at the individual or at the herd level

Valutazione della produzione di gamma interferone in bovini vaccinati con Brucella abortus ceppo RB51 mediante un test ELISA

In questo lavoro sono presentati i risultati di un test ELISA messo a punto per rilevare la produzione di gamma interferone (g-interferone) in bovini vaccinati con Brucella abortus ceppo RB51 (RB51). Come stimolo antigenico per il sangue intero è stata utilizzata una frazione proteica purificata derivante da RB51 (brucellina RB51). La prova è stata valutata nell’arco di 300 giorni in 10 manze vaccinate in età prepubere con 10×109 Unità Formanti Colonia di RB51 e in cinque manze di controllo, provenienti da allevamenti ufficialmente indenni da brucellosi bovina. I capi vaccinati hanno cominciato a fornire risultati positivi a partire dal 17° giorno post vaccinazione (p.v.) fino al giorno 239 p.v. Tutti i capi vaccinati hanno fornito almeno una volta un risultato positivo (indice di stimolazione, IS, superiore a 2,5). Tuttavia, se si esclude il prelievo al giorno 20 p.v. (90% di animali vaccinati risultati positivi), la sensibilità del test oscilla tra il 20% e il 70%, con una media del 40%. IS superiore a 2,5 è stato rilevato anche in tre animali di controllo. Sulla scorta dei risultati ottenuti, si ritiene che il test del g-interferone non fornisce garanzie sufficienti per consigliarne l’impiego ai fini di riconoscere i bovini vaccinati con RB51, sia come prova individuale, sia come prova d’allevamento

Detection of Listeria monocytogenes in food samples in Avezzano, Sulmona and Castel di Sangro (province of L'Aquila, Abruzzo, Italy) between 2000-2009

The retrospective study of the results of the analysed samples is a fundamental tool for the identification of major risk related to food and for planning future monitoring activities. The evaluation of the quality of data collected may also allow for estimating the effectiveness of the controls so to improve their efficacy. In this article, the authors evaluated the results of tests for the detection of Listeria monocytogenes performed by the Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale' (IZSAM) on food samples collected during the years 2000-2009 in the territory of Avezzano, Sulmona and Castel di Sangro (province of L'Aquila, Abruzzo, Italy). The comparison of the data examined with those from studies conducted in Italy and in other countries shows that the categories with higher percentages of positivity for Listeria monocytogenes are meat and fish products. Data collected do not indicate cheese as a vehicle of contamination in the sampled areas, in contrast to what reported in the national and international literature. It would therefore be necessary to promote an ad hoc sampling in the areas covered by this study to verify this aspect in more depth

Using scientific evidence to inform public policy on the long distance transportation of animals: role of the European Food Safety Authority

The authors review the work of the previous Scientific Committee on Animal Health and Animal Welfare and the current European Food Safety Authority (EFSA) in providing scientific advice on the welfare aspects of animal transport and the impact of this advice on the European Union (EU) regulatory framework. Through its Protocol on the Protection and Welfare of Animals, the Treaty of Amsterdam obliges European institutions to pay full regard to the welfare requirements of animals when formulating and implementing EU legislation. Regulation 1/2005 states that EU legislation should be amended to take into account new scientific evidence. Provisions for poultry, cats and dogs take into account the recommendations included in EFSA's Scientific Opinion which considers different species (poultry, deer, rabbits, dogs and cats, fish and exotic animals). Examples of the effect of the scientifically based conclusions and recommendations from the Scientific Opinion on the measures in Regulation 1/2005 are summarised and show the impact of scientific evidence on EU legislation

Isolamento di Brucella suis biovariante 2 da un cinghiale in Abruzzo, Italia

Un cinghiale selvatico femmina, di circa due anni di età, è stato trovato morto dai Servizi Veterinari a Pianola di Roio a L'Aquila, Provincia situata nella Regione Abruzzo nell'Italia centrale. La carcassa è stata conferita all'Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale" di Teramo per l'esecuzione dell'esame autoptico. Dai linfonodi sotto-mandibolari di questo esemplare è stato isolato un ceppo di Brucella suis biovariante 2. Questa è la prima segnalazione d'isolamento di B. suis nella Regione Abruzzo. Diversi autori hanno accettato, in passato, l'ipotesi che B. suis biovariante 2 sia stata introdotta in Italia attraverso l'importazione di lepri provenienti dai paesi europei in cui l'infezione è endemica nelle popolazioni selvatiche. Questa considerazione ha portato le autorità italiane a rafforzare i controlli esistenti sulle lepri selvatiche importate a scopo di ripopolamento. Tuttavia, attualmente, non è in vigore alcuna disposizione (né lo è stata in passato) per il controllo della brucellosi nei cinghiali movimentati, sia a livello nazionale che europeo. L'isolamento di B. suis biovariante 2 da cinghiali in altre Regioni italiane geograficamente distanti potrebbe suggerire che questa specie, piuttosto che le lepri importate, possa essere stata la fonte d'introduzione dell'infezione in tali aree. Le norme nazionali ed europee di gestione della brucellosi nella fauna selvatica dovrebbero essere indirizzate al controllo dello stato di salute dei cinghiali negli allevamenti prima delle movimentazioni o del rilascio, con l'obiettivo di prevenire la diffusione di questo patogeno in territori indenni

Risk Factors for the Spread of Brucellosis in Sheep and Goats in the Campania Region in the Years 2015–2020

Brucella is a Gram-negative facultative intracellular pathogen that causes infection in sheep and goats (B. melitensis.); B. melitensis can also infect other animals. Sheep and goat brucellosis is still present in some regions of Italy, including Campania, and causes considerable economic losses and health threats. The aim of this study was to evaluate the possible risk factors influencing the spread of brucellosis among sheep and goat farms in the Campania region in order to provide the local veterinary services with practical support in evaluating and planning diagnostic, preventive and control interventions. The results of official controls for brucellosis carried out from 2015 to 2020 in the sheep and goat farms of the Campania Region were analyzed. Data were extracted from the National Veterinary Information Systems and the Laboratory Management System of the Experimental Zooprophylactic Institute of Southern Italy. Statistical analysis was carried out through the software R version 4.1.0; the dataset consisted of 37,442 observations, and 9 qualitative and quantitative variables were evaluated on 8487 farms, 248 of which were positive. The association between covariates and the outcome (presence/absence of the disease) was evaluated (Fisher and Wilcoxon tests). A logistic regression model with mixed effects was carried out. This study confirmed that brucellosis in sheep and goats in the Campania region mostly occurs through contact with infected animals imported from other farms (OR = 3.41—IC 95% [1.82–6.41]). Farms with a greater number of animals were seen to be at the greatest risk of infection (OR = 1.04—IC 95% [1.03–1.05]); previous suspension of healthy status also proved to be a risk factor (OR = 55.8—IC 95% [26.7–117])