Infectious Bovine Rhinotracheitis Post-Eradication Program in the Autonomous Province of Bolzano, Italy: A Retrospective Study on Potential Bovine Herpesvirus Type 2 Cross-Reactivity.

Bovine alphaherpesviruses, BoAHV, can cause respiratory, genital and neurological disorders. In particular, bovine alphaherpesvirus type 1 (BoAHV1) is one of the most significant ruminant pathogens worldwide and it can heavily damage the livestock industry. BoAHV1 can cause infectious bovine rhinotracheitis (IBR) along with fertility disorders. Bovine alphaherpesvirus type 2 (BoAHV2) can cause two different conditions as well: pseudo-lumpy skin disease (PSLD) and bovine herpetic mammillitis (BHM). The autonomous province of Bolzano (Italy) has adopted several strategies to control and eradicate IBR, and it was declared in 2000 to be IBR-free by the European Commission. Since 2001, a post-eradication monitoring program has overseen the serological analysis of bulk milk and, in the presence of a positive result, a follow-up examination is performed on the individual blood serum of all bovines older than 24 months that belong to bulk milk-positive herds. Despite the detection of positives in both bulk milk and serum samples, South Tyrol has been declared IBR-free, as these positives have never been confirmed through seroneutralization. Between 2014 and 2022, approximately 41,000 bulk milk (averaging 4300 samples/year) and 3229 serum samples were tested for BoAHV1. The aim of this study was to evaluate the post-eradication program for IBR with a particular focus on the potential cross-reactivity with BoAHV2; for this reason, serum samples were also tested for BoAHV2 antibodies. This study could be of great importance for those countries that submit herds to an IBR monitoring and eradication program; performing further analyses to confirm and explain false positive outcomes would increase the reliability of the obtained results.

Coxiella burnetii outbreak, Northern Italy 2021.

Objective: We report an outbreak of 14 cases of Q fever among tourists caused by an infected calf and characterized by respiratory transmission and a high attack rate in humans. Materials and methods: Following the identification of an index case of Q fever in September 2021, an epidemiological investigation was conducted in collaboration with local Health and Veterinary authorities and an unknown outbreak was discovered and terminated. Results: The outbreak originated from the delivery by an infected cow, with spread of C. burnetii by air and infection via the respiratory route. The transmission period was calculated, and 25 potentially exposed tourists were identified: 14 were infected (56%) based on serological investigations, four were hospitalized, there were no deaths. All the 22 cows were tested by PCR for C. burnetii: 3 cows (14%) were positive on milk samples and one, the index animal, was also positive on blood. Conclusions: Timely diagnosis in a human patient was pivotal to identify the outbreak since involved animals were asymptomatic. The close collaboration between veterinary and human Public Health services in six different geographical areas of two countries was crucial for the rapid termination of the outbreak.

A Seroepidemiological Survey of Corynebacterium pseudotuberculosis Infection in South Tyrol, Italy.

Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis, a bacterial infection that can affect livestock. This infection can cause low growth rates and milk yields and reproductive failure, along with the infection of humans, especially those in close contact with the animals. The aim of this study was to evaluate the local CLA prevalence, highlighting the parameters for the potential predisposition to infection in goats, and to implement a monitoring program based on the newly acquired scientific evidence. Of a total of 2365 goat farms in South Tyrol, 384 farms were selected for the present study. A statistically significant number of animals were subjected to serologic analysis for the detection of C. pseutotubercolosis antibodies. A total of 2948 goats belonging to the selected farms were tested for CLA, 713 of which showed a positive result. The data analysis led to an estimated CLA seroprevalence of 21.85%. The results achieved can enable the evaluation and possible implementation of a voluntary program that permits us to test a larger number of animals using serological techniques. This program would be of great importance, especially for those farms dedicated to the production of milk and dairy products, as some manufacturing practices may increase the risk of transmission of zoonotic pathogens such as C. pseudotuberculosis to humans.

Risk Factors Associated with the Alpine Multispecies Farming System in the Eradication of CAEV in South Tyrol, Italy.

South Tyrol has implemented, in 2007, a mandatory eradication program against Caprine Arthritis Encephalitis Virus (CAEV), a virus known to cause economic losses related to decreases in milk production and milk quality in goats, along with poor animal welfare and premature death. After a great initial decrease in the seroprevalence, the program has reached a tailing phase with scattered positivities. Potential risk factors associated with the multispecies farming system, a traditional approach in South Tyrol, are evaluated in this study, in order to better understand some of the potential causes leading to the tailing phenomenon. A statistically significant number of farms was selected for the present study, based on the risk factors evaluated. Even though there is no statistically significant association between the practices evaluated and the incidence of infection, the authors believe that it is important to highlight potential risks that may threaten the outcome of this eradication program.

Business intelligence tools to optimize the appropriateness of the diagnostic process for clinical and epidemiologic purposes in a multicenter veterinary pathology service.

Laboratory tests provide essential support to the veterinary practitioner, and their use has grown exponentially. This growth is the result of several factors, such as the eradication of historical diseases, the occurrence of multifactorial diseases, and the obligation to control endemic and epidemic diseases. However, the introduction of novel techniques is counterbalanced by economic constraints, and the establishment of evidence- and consensus-based guidelines is essential to support the pathologist. Therefore, we developed standardized protocols, categorized by species, type of production, age, and syndrome at the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), a multicenter institution for animal health and food safety. We have 72 protocols in use for livestock, poultry, and pets, categorized as, for example, "bovine enteric calf", "rabbit respiratory", "broiler articular". Each protocol consists of a panel of tests, divided into 'mandatory' and 'ancillary', to be selected by the pathologist in order to reach the final diagnosis. After autopsy, the case is categorized into a specific syndrome, subsequently referred to as a syndrome-specific panel of analyses. The activity of the laboratories is monitored through a web-based dynamic reporting system developed using a business intelligence product (QlikView) connected to the laboratory information management system (IZILAB). On a daily basis, reports become available at general, laboratory, and case levels, and are updated as needed. The reporting system highlights epidemiologic variations in the field and allows verification of compliance with the protocols within the organization. The diagnostic protocols are revised annually to increase system efficiency and to address stakeholder requests.

Koi herpesvirus disease outbreak: Input for the implementation of a surveillance program in South Tyrol - Italy.

A severe episode of high and abnormal mortality was observed in the population of Cyprinus carpio of Lake Caldaro (South Tyrol, Italy) in summer 2016. The diagnostic investigation carried out led to the identification of Koi Herpesvirus (KHV) as the etiologic agent. Following this disease outbreak and its socio-economic consequences, the local authorities, in agreement with the local Fishing Association, decided to implement a surveillance program for the achievement of KHV-free health status (Category I) in the Province, in accordance to the Implementing Decision (EU) 2015/1554. The selected area was a defined geographical compartment (the Monticolo lakes compartment, South Tyrol, Italy), which is located near Lake Caldaro, where the Koi Herpesvirus disease (KHVD) outbreak had occurred. This area is of particular interest because it supplies other water bodies with juvenile C. carpio individuals; with the achievement of a KHV-free health status, South Tyrol could possibly become independent in the breeding of this fish species. Suitable samples were collected and processed during a two-year period in order to detect the presence/absence of KHV. The same samples were tested for other viruses that can affect carp, namely spring viraemia of carp (SVCV) and carp edema virus (CEV). According to the results, the authors conclude that the Monticolo lakes area should be classified as KHV-free, as no sample has tested positively for the presence of this specific virus (KHV).

Eradication of caprine arthritis encephalitis virus in the goat population of South Tyrol, Italy: analysis of the tailing phenomenon during the 2016-2017 campaign.

Since 2007, the Autonomous Province of Bolzano-South Tyrol (Italy) has carried out a compulsory eradication program against caprine arthritis encephalitis virus (CAEV) in goats. A drastic seroprevalence reduction was achieved during the initial phase (2007-2011); however, a tailing phenomenon has been observed during the latest years, hampering the achievement of the final goal. CAEV belongs to a group of lentiviruses, called small ruminant lentiviruses (SRLVs), which are antigenically related and can infect both goats and sheep. We investigated the possible link between the tailing phenomenon in goats and the role of sheep as a virus reservoir by comparing serologic results between multispecies farms (where goats and sheep coexist) and monospecies farms (goats only). Goats on multispecies farms had a higher prevalence and seroconversion rate (even if to a rather moderate extent), higher antibody titers, and a higher probability of conclusive results in the genotyping analysis, with more frequent identification of SRLV genotype A (sheep-related) infections. Sheep can serve as a SRLV reservoir, thus contributing to scattered positive tests in goats, causing the tailing phenomenon.

Use of a commercial ELISA kit specific for glycoprotein E peptides to indirectly detect Caprine Herpesvirus 1 (CpHV-1) in the state of São Paulo, Brazil / Uso de kit ELISA comercial específico para peptídeos da glicoproteína E para detecção indireta de Herpesvírus Caprino tipo 1 (CpHv-1) no estado de São Paulo, Brasil

Caprine herpesvirus 1 (CpHV-1) infection is associated with clinical manifestations related to animal age, with high mortality in kids and infertility in adults. Given the scarcity of research about the epidemiological situation of this infection in Brazilian flocks, we aimed to conduct a cross-sectional descriptive study to detect antibodies against CpHV-1 in goats in the state of São Paulo, Brazil. Fifty-five male and female goats ­ kids and adult ­ were assessed in this study. Blood serum was analyzed by a commercial ELISA kit to detect antibodies against CpHV-1, which had not been used in Brazil before. No animals were reactive. Brazil lacks information about CpHV-1 infection in goat flocks. Continuing the study is crucial to understand the epidemiological situation of the disease and establish protocols for infection control.(AU)

A infecção pelo Herpesvírus Caprino tipo 1 (CpHv-1) está associada a manifestações clínicas relacionadas à idade dos animais, com alta mortalidade em filhotes e infertilidade em adultos. Diante da escassez de estudos sobre situação epidemiológica dessa infecção nos rebanhos brasileiros, a presente pesquisa teve como objetivo realizar um estudo transversal e descritivo para a detecção de anticorpos anti-Herpesvírus Caprino tipo 1 em caprinos do estado de São Paulo, Brasil. Foram avaliados 55 caprinos machos e fêmeas, filhotes e adultos. O soro sanguíneo foi analisado por um kit ELISA comercial para detecção de anticorpos contra CpHv-1, de utilização inédita no Brasil. Nenhum animal estudado foi sororreagente. O Brasil carece de informações acerca da infecção pelo Herpesvírus Caprino tipo 1 nos rebanhos caprinos do país. A continuidade do estudo é imprescindível para compreender a situação epidemiológica da enfermidade e estabelecer protocolos para controle da infecção.(AU)

Isolation of Streptococcus agalactiae in a female llama (Lama glama) in South Tyrol (Italy).

BACKGROUND: Streptococcus agalactiae is pathogenic for both animals and humans. In dairy cattle it commonly causes mastitis, with great economic losses, and there is scientific evidence of mastitis, caseous lymphadenitis, contagious skin necrosis and purulent infections associated with S. agalactiae in camels (Camelus dromedarius) as well. In humans, it is a common component of the respiratory and gastrointestinal microflora, but it can also act as a pathogen, especially in elderly people and immunocompromised patients, as well as in pregrant women and newborns. CASE PRESENTATION: A 10-year old non-pregnant female llama (Lama glama) was conferred to the Institute for Animal Health Control, in Bolzano for necropsy after sudden death. The animal had not shown unusual behaviour and had a low to normal nutritional condition (body condition score 2/5). The breeder had reported a chronic suppurative subcutaneous infection in the intermandibular area, resistant to therapy (therapy unknown). After necropsy, several samples were processed for histological, bacteriological and parasitological examinations. CONCLUSIONS: This report describes, to the best of our knowledge, the first isolation of S. agalactiae in llamas (Lama glama). The animal came from a herd that counts approximately 200 South American camelids (llamas, alpacas) along with several horses, chicken, rabbits, cats and dogs; this farm offers services, such as trekking and pet therapy activities.

Achievements of an eradication programme against caprine arthritis encephalitis virus in South Tyrol, Italy.

Small ruminant lentivirus infections in goats affect both production and animal welfare. This represents a threat to the qualitative and quantitative growth of goat farming, recently observed in mountainous regions such as the Autonomous Province of Bolzano - South Tyrol (Italy). To monitor and eradicate the caprine arthritis encephalitis virus in this goat population, a compulsory eradication campaign was launched, based on a strict census of small ruminants and yearly serological testing of all animals, followed by the consequent culling of seropositive individuals. The campaign succeeded in completely eliminating cases of clinical disease in goats, while drastically reducing the seroprevalence at the herd as well as individual animal level. The serological outcome of the introduced control measures was determined using commercially available ELISA kits, demonstrating their suitability for use in this type of campaign, aimed at reducing seroprevalence as well as clinical manifestations of these infections. However, this clear success is diminished by the failure to achieve a complete eradication of these viruses. The reasons leading to the observed tailing phenomenon and the occurrence of new infections in already sanitised flocks are discussed and implementation of further measures are proposed.

Dystrophic mineralization of the arterial fibrovascular tissue associated with a vitamin D hypervitaminosis in an 8-year-old female Alpaca (Vicugna pacos).

BACKGROUND: Prophylactic Vitamin D supplementation is a common practice in Alpaca breeding in many regions around the world. An overdosage can lead to dystrophic mineralization of soft tissues. In this paper we illustrate a suspected case of hypervitaminosis D in an 8-year-old female Alpaca. CASE PRESENTATION: In June 2015, the carcass of an 8-year-old female Alpaca (Vicugna pacos) was submitted to the diagnostic laboratory of the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe) for necropsy. The animal had undergone a spontaneous abortion with uterine prolapse and delivery of the placenta, and had died shortly thereafter. Death occurred due to internal haemorrhage related to dystrophic mineralization of the left renal artery with subsequent rupture and damage of the renal hilum. During the necropsy, histopathological and serum biochemical analyses were performed. After laboratory analyses and the history of mineral and vitamin supplementation reported by the breeder, a hypervitaminosis D was suspected to be the most probable cause of the dystrophic mineralization observed in the left renal artery. CONCLUSIONS: Most of the information regarding Llamas and Alpacas comes from the South American and Australian regions. It is therefore important to provide scientific information about these animals in other regions of the world in order to have a better and wider understanding of the nutritional and environmental conditions necessary for optimal breeding.

Evidence for a natural humoral response in dairy cattle affected by persistent botulism sustained by non-chimeric type C strains.

Bovine botulism is a sporadic acute disease that usually causes catastrophic losses in the herds. The unusual clinical evolution of a persistent mild outbreak in a dairy herd, prompted us to characterize the neurotoxin gene profile of the strain involved and to evaluate whether seroconversion had occurred. Diagnosis was based on mild classical symptoms and was supported by PCR and bacteriological findings, which revealed the involvement of a non-mosaic type C strain. An in-house ELISA was developed to detect antibodies to botulinum neurotoxin type C and its performance was evaluated in a vaccination study. Fifty days after the index case, fecal and serum samples were collected from the 14 animals of the herd and screened for Clostridium botulinum and anti-botulinum neurotoxin antibodies type C, respectively. The in-house developed ELISA was also used to test 100 sera samples randomly collected from 20 herds. Strong ELISA reactions were observed in 3 convalescent and 5 asymptomatic animals involved in the studied outbreak. The ELISA-positive cows all tested positive for non-mosaic C. botulinum type C in the feces and the same strain was also detected in the alfalfa hay, suspected to be the carrier source. Ten out of the 100 randomly collected sera tested positive for anti-botulinum neurotoxin type C antibodies: 7 had borderline values and 3 from the same herd showed titers three times higher than the cut-off. We concluded that type C botulism in cattle may occur with variable severity and that prolonged exposure to sublethal doses of botulinum neurotoxin C may occur, resulting in detectable antibodies.

[Retrospective evaluation of data over five years of the BVDV prevention program using ear notch samples from the autonomous province of Bolzano (Italy)]. / Retrospektive Auswertung von Daten über fünf Jahre BVDV-bekämpfungsprogramm mittels Ohrgewebeproben in der Autonomen Provinz Bozen (Italien).

In 2005 the Autonomous Province of Bolzano implemented a BVD-Virus control programme by testing all newborn calves for Bovine Viral Diarrhoea Virus (BVDV) antigen by analysing ear notch samples. In eradication programs between November 2005 and October 2010 a total of 344,108 newborn calves were ear notch sampled and analysed for BVDV by an antigen ELISA detecting the E(rns) (HerdCheck BVDV Antigen/Serum Plus, Idexx Laboratories, Switzerland). The tissue samples were collected during calf ear tagging (within the first 20 days of life) using two sampling devices (TypiFix-System, Agrobiogen GmbH [Germany] and Allflex [France]). 0.4% (1400) of the collected samples showed a positive result, and of these, 583 calves were subjected to a follow-up examination, analysing the samples again by ELISA as well blood samples by Reverse transcriptase-PCR. Blood samples were also collected from the dam, if still present on the farm by the time of sampling. These results suggest that a BVDV control programme testing all newborn calves sampled for BVDV antigen by analysing an ear notch sample in principle is practicable. However, due to the low positive predictive value (75%) of the E(rns)-ELISA used, not all persistently infected calves can be detected.