Calves gastrointestinal nematodes and Eimeria prevalence and associated risk factors in dairy farms, southern Ethiopia

Erratum to: Calves gastrointestinal nematodes and Eimeria prevalence and associated risk factors in dairy farms, southern EthiopiaMaireg Hailu1, Kassahun Asmare2, Endrias Zewdu Gebremedhin3, Vincenzo Di Marco Lo Presti4, Maria Vitale4, Desie Sheferaw2On page 6 of the original publication, a short paragraph and a table (the second Table 4) were wrongly included from other article during layout preparation. Therefore, this wrong information has nothing to do with the topic and is deleted from the original publication Abstract:  Dairy production is an important component of livestock farming in Ethiopia. Nevertheless, the productivity of the sector has been impacted negatively by the morbidity and mortality of replacement animals. A Cross-sectional study was, therefore, aimed at estimating the prevalence of Nematode and Eimeria infection in calves in Hawassa, Shashemene and Arsi Negelle, southern Ethiopia. To this end, a flotation technique was used to recover Nematode egg and Eimeria oocyst from rectally collected faeces. The overall prevalence of gastrointestinal parasitic infection, Nematode and Eimeria species collectively, was 43.9% (95% CI=38.6-49.4). The estimated proportion of Nematode, Eimeria and mixed infection was 35.8%, 21.5% and 13.3%, respectively. Among the potential factors considered faecal consistency, age and study area were found to increase recovery of Nematode egg and Eimeria oocyst in faeces (p< 0.05). Area wise, the prevalence has been noted to be higher at Arsi Negelle followed by Hawassa and Shashemene (p< 0.05). Besides, younger and diarrheic calves were found more infected by Nematode and Eimeria species than their adult and non-diarrheic counterpart. Based on their morphological appearance, Strongyle type (20.3%), Trichuris (4.2%) and Ascaris (16.3%) eggs, Eimeria oocysts (21.5%) were observed. In the light of this finding, the authors would like to advise the need for strategic intervention

Multiple drug-susceptibility screening in Mycobacterium bovis: new nucleotide polymorphisms in the embB gene among ethambutol susceptible strains

Objectives: Pyrazinamide-resistant Mycobacterium bovis isolates of animal origin were assessed for drug susceptibility to five antituberculosis drugs by the agar based Middlebrook 7H11 method as gold standard as well as by a simplified, dichotomous resazurin microtitre assay (d-REMA). Methods: A total of 53 M. bovis isolates were typed and tested against isoniazid, rifampin, streptomycin, ethambutol, kanamycin and the control drug pyrazinamide. On the basis of the results obtained, pncA and embB genes were PCR-amplified and DNA-sequenced for all isolates. Results: All M. bovis isolates, classified into 21 spoligotype/MIRU-VNTR profiles, were resistant to pyrazinamide by both methods, as expected. The pncA gene sequencing confirmed the presence of the resistance-conferring H57D mutation. All strains were found to be susceptible to the other five drugs by the agar based gold standard method. The d-REMA was in agreement with these results for all five drugs, with the exception of 12 isolates, which showed ambiguous and therefore inconclusive results in ethambutol testing. Mutations in the embB gene were observed in all 53 isolates: four new single-nucleotide polymorphisms were identified. No association was found between embB genetic profiles and ethambutol resistance results by the gold standard. Conclusion: All M. bovis isolates were sensitive to the most common antituberculosis drugs used for treatment. There was a good agreement between the d-REMA assay and the agar based reference method. Among ethambutol susceptible isolates, four new embB mutations were found

Antibiotic Resistance Profiling, Analysis of Virulence Aspects and Molecular Genotyping of Staphylococcus aureus Isolated in Sicily, Italy

Staphylococcus aureus is the major cause of foodborne diseases worldwide. In this retrospective study, 84 S. aureus strains were characterized. The collection comprises 78 strains isolated during 1998 and 2014 from dairy products and tissue samples from livestock bred for dairy production in Sicily. One isolate was obtained from a pet (dog), one from an exotic animal (a circus elephant), and four human isolates were obtained during a severe food poisoning outbreak that occurred in Sicily in 2015. All the strains were characterized by pulsed-field gel electrophoresis (PFGE), for antibiotic resistance and presence of toxin genes. PFGE results showed 10 different pulsotypes, with three relatively frequent and three unique. The antibiotic resistance profiling showed that penicillin G (35.7%) and tetracycline (20.2%) resistance is largely spread. Most isolates contained at least one toxin gene making them a potential threat for public health. Enterotoxin sec gene was observed in 28.6% and seg in 23.8% of the strains, respectively; the human isolates were the only ones to concurrently harbor both seg and sei genes. In addition, 24 isolates were randomly selected and analyzed by multilocus sequence typing. Interestingly, the analysis showed the presence of 12 sequence types (STs), of which 6 were novel. One of them, ST700, was detected in 29% of the isolates and was found to be spread throughout Sicily. ST700 has been present in the island for almost 16 years (1998-2014) and it shows no host preference since it was isolated from different ruminant species. Four human isolates shared both the pulsotype (PT10) and the sequence type (ST9), as well as the virulence genes (seg-sei); this observation suggests that the isolates originated from a single clone, although they were obtained from two different individuals

Comparison of semi-automated commercial rep-PCR fingerprinting, spoligotyping, 12-locus MIRU-VNTR typing and single nucleotide polymorphism analysis of the embB gene as molecular typing tools for Mycobacterium bovis

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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

Inside Mycobacterium bovis SB0120 spoligotype circulating in Italy: analysis of the most frequent genotypes by whole genome sequencing

Bovine tuberculosis (bTB) is a chronic inflammatory disease primarily caused by Mycobacterium bovis. The infection affects domestic animals and wildlife, posing a zoonotic risk to humans. To understand the dynamics of transmission and genetic diversity in Italy’s M. bovis population, we conducted whole-genome sequencing (WGS) analysis on two prevalent genotypes, belonging to Spoligotype SB0120, identified in different geographical and temporal contexts. By comparing these genomes with international M. bovis isolates, we identified a distinct clade within the lineage La1.2, encompassing the Italian SB0120 isolates, indicating a genomic segregation of Italian M. bovis from other European isolates. Within Italy, a significant level of genetic variability emerged across regions, while isolates within epidemiologically linked outbreaks exhibited minimal genetic diversity. Additionally, isolates derived from cattle and wild boars within a tuberculosis hotspot in Central Italy and from cattle and black pigs in Sicily formed unified clonal clusters. This indicates the presence of persistent strains circulating in the examined regions. The genetic diversity within herds was limited, as specific clones endured over time within certain herds. This research enhances our comprehension of the epidemiology and transmission patterns of bTB in Italy, thereby aiding the development of precise control strategies and disease management. Using WGS and implementing standardized protocols and databases will be pivotal in combating bTB and promoting One-Health approaches to address this noteworthy public health concern