Doppler sonography of the corpus luteum during the oestrus cycle in dairy cows

Doppler ultrasound has recently emerged as one of the main innovations in cattle practice and has mainly found application in the evaluation of luteal blood perfusion (LBF). LBF has been evaluated during different phases of the oestrous cycle, but the technique is poorly standardized. The aim of the study was to evaluate visual and quantitative changes of size of corpus luteum (CL) and LBF in dairy cattle during an oestrous cycle at 2 follicular waves using colour flow mode (CFM) and power flow mode (PFM) doppler ultrasound. Ten Friesian cows were selected and synchronized. The CL was evaluated at 3/4-day intervals until the next oestrus onset. After identifying the spiral luteal artery entry at the base of the CL, multiple scans of the vertical plane at the maximum diameter of the CL from the apex to the base were recorded in B-mode, CFM and PFM for each cow and session. An Esaote Mylab vet 30 gold was employed with the following setting: pulse repetition frequency 2.1 (CFM) or 2.8 (PFM), gain 70%. The real area of CL (RACL) was calculated subtracting the area of the eventual inner cavity. The LBF was quantified off-line by means of a visual score and of an image analysis system (Digimizer 4.1). Student and Anova tests were used for statistical analysis. Data were presented as means and standard deviations. There were significant individual variations for the RACL and LBF when calculated in CFM. PFM minimized these variations, although, at day 15-16, it was also affected by individual factors (p < 0.01). The combined study of RACL and LBF during a 2-wave oestrous cycle distinguished 3 phases: a phase of CL formation (3-4 days) with LBF of 0.3 ± 0.3 cm2 on a RACL of 3.1 ± 0.9 cm2, a central phase (7-16 days) with LBF of 0.7 ± 0.4 cm2 on a RACL of 4.8 ± 0.8 cm2 and a regression phase (19-20 days) with a minimum LBF of 0.1 ± 0.1 cm2 in a CL of 3.3 ± 0.6 cm2. According to this study, there is no significant difference of LBF from 7 to 16 days of the oestrus cycle; although the RACL significantly changes in this period, the dimensional variations are too thin to be clinically appreciated. However, this study contributed to defining the values of RACL and LBF during a 2-wave oestrous cycle of dairy cattle

The detection of anti-Trichinella antibodies in free-ranging Nebrodi Regional Park black pigs from Sicily, Italy, suggests the circulation of Trichinella britovi in the island

Abstract • Zoonotic nematodes of the genus Trichinella are known to occur in countries bordering the Mediterranean Sea. For example, Trichinella spiralis was reported in pigs and humans at the turn of the Second World War in Sicily and Trichinella britovi in pigs, red foxes and dogs from Corsica and Sardinia since 2004. Following the discovery of T. britovi in Corsica and Sardinia, a question arose as to whether this species was also present in Sicily which is located only 3.14 km from continental Italy. To address this query, we investigated the presence of anti-Trichinella antibodies in the serum of Nebrodi black pigs, a breed that is bred in the wild in the Nebrodi Regional Park, a protected area of the island rich in flora and fauna. Blood samples were collected from 112 Nebrodi black pigs from five farms. Sera were tested by ELISA and ELISA positive sera were confirmed by Western blot (Wb) using excretory/secretory antigens. Eighteen (16.1%) serum samples belonging to 17 fattening pigs and 1 boar tested positive by Wb. Positive sera tested by Wb using crude worm extract antigens (CWE), displayed a banding pattern similar to the CWE-Wb pattern of T. spiralis and T. britovi reference pig sera but different to that of T. pseudospiralis reference pig sera. No larvae were detected in muscles of serologically positive pigs by artificial digestion. The presence of anti-Trichinella antibodies in the absence of larvae in the muscles, suggests that the pigs were infected with T. britovi and not T. spiralis whose larvae survive in the muscles for at least two years. These results suggest that T. britovi is circulating in Nebrodi Regional Park in Sicily. ELISA testing may constitute a suitable tool for large-scale screening of Trichinella spp. infection in free-ranging pigs, when ELISA-positive sera are confirmed by Wb. Free-ranging animals can act as sentinels for the presence of zoonotic nematodes of the genus Trichinella in wildlife

Geo-epidemiology of animal tuberculosis and Mycobacterium bovis genotypes in livestock in a small, high-incidence area in Sicily, Italy

IntroductionThe persistence of animal tuberculosis (TB) in livestock is a major concern in Sicily, Italy. The objective of this study was to elucidate the transmission dynamics of M. bovis infection in a highly circumscribed, and at the same time geographically diverse, high-risk area of the island through an in-depth geo-epidemiological investigation of TB in cattle and black pigs raised in small-scale extensive farms across the district of Caronia.MethodsWe used genotype analysis coupled with geographic information system (GIS) technology and phylogenetic inference to characterize the spatial distribution of TB and M. bovis genotypes in livestock and the genetic relationships between M. bovis isolates. A total of 589 M. bovis isolates collected from slaughtered cattle (n = 527) and Sicilian black pigs (n = 62) over a 5-year period (2014–2018) were included in the study.ResultsTB was widespread throughout the district and was most frequent in the north-central area of the district, especially along one of the district’s streams. We identified a total of 62 M. bovis genotypes. Identical genetic profiles were isolated from both neighboring and non-neighburing herds. The 10 most frequent genotypes, accounting for 82% of M. bovis isolates, showed geographic specificities in that they tended to cluster in specific spatial niches. The landscape structure of these niches—i.e. steep slopes, rocky ridges, meadows and streams—is likely to have had a significant influence on the distribution of TB among livestock in Caronia. Higher concentrations of TB were observed along streams and in open meadows, while rocky ridges and slopes appeared to have hampered the spread of TB.DiscussionThe geographical distribution of TB cases among livestock in Caronia is consistent with several epidemiological scenarios (e.g., high density of infected herds along the streams or in hilly plateau where livestock share pastures). Landscape structure is likely to play an important role in the transmission and persistence of M. bovis infection across the district. Additional potential risk factors, such as livestock trading and extensive breeding methods, are also discussed. Our results will contribute to the improvement of surveillance, control and eradication activities of TB in Sicily by the implementation of ad hoc TB control measures, especially in farms located along streams, sharing common pastures or with mixed animal species

Genotype diversity and distribution of Mycobacterium bovis from livestock in a small, high-risk area in northeastern Sicily, Italy.

Bovine tuberculosis (bTB) caused by Mycobacterium bovis is an important re-emerging disease affecting livestock, wildlife and humans. Epidemiological studies are crucial to identifying the source of bTB infection, and its transmission dynamics and host preference, and thus to the implementation of effective strategies to contain it. In this study, we typed M. bovis isolates from livestock, and investigated their genetic diversity and distribution. A total of 204 M. bovis isolates were collected from cattle (n = 164) and Sicilian black pigs (n = 40) reared in a limited area of the province of Messina, northeastern Sicily, an area that had previously been identified as having the highest incidence of bTB in livestock on the island. All M. bovis isolates were typed by both spoligotyping and 12-loci MIRU-VNTR analysis. Results from both methods were then combined in order to improve the discriminatory power of M. bovis typing. We identified 73 combined genetic profiles. Thirty-five point six percent of the profiles were common to at least two animals, whereas 64.4% of profiles occurred in only one animal. A number of genetic profiles were predominant in either cattle or black pigs. We identified common genetic patterns in M. bovis isolates originating not only from neighboring districts, but also from non-neighboring districts. Our findings suggest that bTB is widespread in our setting, and is caused by a large number of genetically diverse M. bovis strains. The ecology and farming practices characteristic of the area may explain the substantial M. bovis heterogeneity observed, and could represent obstacles to bTB eradication

Aujeszky’s disease in hunting dogs after the ingestion of wild boar raw meat in Sicily (Italy): clinical, diagnostic and phylogenetic features

Abstract Aujeszky's disease is caused by Suid Herpes Virus-1 and species belonging to the genusSus scrofaare the main reservoir hosts. This virus, however, is capable of infecting and causing severe disease, with an almost constant fatal outcome in other species, both domestic and wild (carnivores, monogastric herbivores and ruminants). Moreover, the possibility of transmission to humans has been demonstrated. This study reports and describes the clinical, diagnostic, pathological and phylogenetic aspects of two cases of Aujeszky's disease in two hunting dogs following the ingestion of infected wild boar raw meat. These cases are contextualized in the province of Messina (Sicily), where a high prevalence of Aujeszky's disease has been recorded (average of 12,20% in the period 2010–2019) in farmed pig, and with evidence of spread to other species. A severe outbreak in cattle has recently been reported in these areas. Nevertheless, cases of Aujeszky's disease in dogs are rarely reported and this study represents the first well-documented report in this species in Sicily.Case presentation&lt;jats:p/&gt;After a wild boar hunt, two dogs showed neurological symptoms and intense itching unresponsive to therapy. Diagnosis of Aujeszky's disease was made based on clinical suspicion, anamnestic information and confirmed by the isolation of the virus from the brain of both dogs. In addition, molecular typing, sequencing and phylogenetic analysis of the Real-Time PCR products were performed. The sequences studied were placed in the Italian Clade 1 along with the sequences obtained from wild boars and hunting dogs from Italy and France.Conclusions&lt;jats:p/&gt;The finding of this disease in non-natural hosts in Sicilian multi-host epidemiological contexts suggests that the risk of inter-species transmission is concrete and that attention should be paid to developing disease control programs in these territories. The data obtained from genome sequencing of the two SuHV-1 isolates contribute to the enrichment of the GenBank with unknown sequences and the phylogenetic analysis implementation

Comparative Study of <i>Mycobacterium bovis</i> and <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> In Vitro Infection in Bovine Bone Marrow Derived Macrophages: Preliminary Results

Bovine tuberculosis and paratuberculosis are endemic in many areas worldwide. This work aims to study cytokines production and gene expression profiles of bovine macrophages infected with Mycobacterium bovis and Mycobacterium paratuberculosis subsp. avium (MAP) strains to identify potential diagnostic biomarkers. Bovine bone marrow stem cells were differentiated into macrophages and subsequently infected in vitro with different spoligotypes of M. bovis and MAP field strains (as single infections and coinfections), using different multiplicity of infection. Supernatant and cell pellets were collected 24 h, 48 h, and one week post-infection. Preliminarily, gene expression on cell pellets of IL-1β, IL-2, INFγ, IL-6, IL-10, IL-12, and TNFα was assessed by qRT-PCR one week p.i. Subsequently, IL-1β and IL-6 were measured by ELISA and qRT-PCR to investigated their production retrospectively 24 h and 48 h p.i. A variability in macrophages response related to the concentration of mycobacteria, the coinfection with MAP, and M. bovis spoligotypes was identified. An early and constant IL-6 increase was observed in the M. bovis infection. A lower increase in IL-1β was also detected at the highest concentration of the two M. bovis spoligotypes one week post-infection. IL-6 and IL-1 β production was reduced and differently expressed in the MAP infection. IL-6 appeared to be the earliest cytokines produced by bovine macrophages infected with M. bovis

Retrieving Historical Cases of Aujeszky’s Disease in Sicily (Italy): Report of a Natural Outbreak Affecting Sheep, Goats, Dogs, Cats and Foxes and Considerations on Critical Issues and Perspectives in Light of the Recent EU Regulation 429/2016

Aujeszky’s disease is caused by Suid alphaherpesvirus 1, and its main reservoir host is the pig. However, other species are also susceptible. Infection with this virus causes a severe neurological clinical picture named Aujeszky’s disease, usually accompanied by itching and death a few days after the onset of symptoms. This study reports a multi-species outbreak of Aujeszky’s disease that occurred in Sicily, which led to the death of 2 goats, 15 sheep, 2 dogs, 2 cats and 2 foxes. The diagnosis was made by culture, indirect immunofluorescence on brain samples and confirmed by biological test on rabbits. This study reports the first cases of Aujeszky’s disease in Italy in cats, goat and sheep. The finding of Aujeszky’s disease in several species in Sicily suggests a potential epizootic risk. In such areas where a multi-host system is recognised, an analysis of the risk factors should be carried out in order to develop targeted strategies for the control and eradication of the disease. The critical issues that hinder the control of Aujeszky’s disease in the studied territory and perspectives for eradication in the light of EU regulation 429/2016 are also discussed.&lt;/jats:p&gt

<i>Mycobacterium bovis</i> Tuberculosis in Two Goat Farms in Multi-Host Ecosystems in Sicily (Italy): Epidemiological, Diagnostic, and Regulatory Considerations

Mycobacterium bovis (M. bovis) is the causative agent of animal tuberculosis (bTB), infecting and causing disease in several animal species. In areas where there are complex interactions between reservoir hosts and susceptible species, the control of this pathogen is a challenge. The authors report two outbreaks of goat tuberculosis caused by M. bovis in multi-host ecosystems within two protected natural areas of Sicily, where TB is historically endemic. The first outbreak (Farm A) was identified after the incidental detection at the slaughterhouse of TB-like lesions in goat viscera ready to be disposed. Single intradermal cervical tuberculin test (SICT) was performed in Farm A on 205 goats, resulting positive in 10 (4.9%). After slaughtering, six out of ten animals showed TB-like lesions, from which M. bovis spoligotype SB0841 was isolated. The typing did not reveal any epidemiological connection with the neighboring cattle, suggesting that free-ranging type of management exposed the affected goat livestock or wildlife infected with other strains. The second outbreak (Farm B) was detected in a mixed farm (bovine, caprine, and ovine), where relapsing outbreaks of TB in cattle were registered in the previous years after performing the SICT in cohabiting goats. SICT resulted positive in 6/153 (3.9%), and two animals showed bTB-like lesions. No mycobacteria were cultured, and the final diagnosis of TB was achieved by histopathology and immunohistochemistry. The reported outbreaks highlight the importance of assessing the epidemiological, diagnostic, and regulatory critical issue, which is fundamental to optimizing the strategies of eradicating TB in the endemic multi-host ecosystem described