Severe Streptococcus equi Subspecies zooepidemicus Outbreak from Unpasteurized Dairy Product Consumption, Italy.

During November 2021-May 2022, we identified 37 clinical cases of Streptococcus equi subspecies zooepidemicus infections in central Italy. Epidemiologic investigations and whole-genome sequencing showed unpasteurized fresh dairy products were the outbreak source. Early diagnosis by using sequencing technology prevented the spread of life-threatening S. equi subsp. zooepidemicus infections.

Recent Trends of Antibiotic Resistance in Staphylococcus aureus Causing Clinical Mastitis in Dairy Herds in Abruzzo and Molise Regions, Italy.

This study aimed to investigate the recent trends of antibiotic resistance (AR) prevalence in Staphylococcus aureus isolated from the milk of animals with clinical mastitis in areas of the Abruzzo and Molise regions in Central Italy. Fifty-four S. aureus isolates were obtained from routine testing for clinical mastitis agents carried out in the author institution in the years 2021 and 2022 and were analyzed for phenotypic resistance to eight antibiotics recommended for testing by European norms and belonging to the antibiotic classes used for mastitis treatment in milk-producing animals. Moreover, the presence of 14 transferable genetic determinants encoding resistance to the same antibiotics was analyzed using qPCR tests developed in this study. Phenotypic resistance to non-ß-lactams was infrequent, with only one 2022 isolate resistant to clindamycin. However, resistance to the ß-lactam cefoxitin at concentrations just above the threshold of 4 µg/mL was observed in 59.2% of isolates in both years, making these isolates classifiable as methicillin-resistant. The AR genotypes detected were the blaZ gene (50% of 2021 isolates and 44.4% of 2022 isolates), aphA3-blaZ- ermC/T (one 2021 isolate), aphA3-ant6-blaZ-ermC/T (one 2021 isolate), blaZ-ermB (one 2022 isolate) and mecA-mph (one 2022 isolate). An inquiry into the veterinarians who provided the samples, regarding the antimicrobials prescribed for mastitis treatment and criteria of usage, indicated a possible causal relation with the AR test results. The occurrence of AR genotypes did not increase in time, most probably reflecting how mastitis was treated and prevented in farms. However, the frequently observed cefoxitin resistance needs to be explained genotypically, further monitored and limited by modifying antibiotic usage practices. The identification of a mecA-positive isolate in 2022 suggests further investigation if this genotype is emerging locally.

Multi-approach methods to predict cryptic carbapenem resistance mechanisms in Klebsiella pneumoniae detected in Central Italy.

The rapid emergence of carbapenem-resistant Klebsiella pneumoniae (Kp) strains in diverse environmental niches, even outside of the clinical setting, poses a challenge for the detection and the real-time monitoring of novel antimicrobial resistance trends using molecular and whole genome sequencing-based methods. The aim of our study was to understand cryptic resistance determinants responsible for the phenotypic carbapenem resistance observed in strains circulating in Italy by using a combined approach involving whole genome sequencing (WGS) and genome-wide association study (GWAS). In this study, we collected 303 Kp strains from inside and outside clinical settings between 2018-2022 in the Abruzzo region of Italy. The antimicrobial resistance profile of all isolates was assessed using both phenotypic and bioinformatic methods. We identified 11 strains resistant to carbapenems, which did not carry any known genetic determinants explaining their phenotype. The GWAS results showed that incongruent carbapenem-resistant phenotype was associated specifically with strains with two capsular types, KL13 and KL116 including genes involved in the capsule synthesis, encoding proteins involved in the assembly of the capsule biosynthesis apparatus, capsule-specific sugar synthesis, processing and export, polysaccharide pyruvyl transferase, and lipopolysaccharide biosynthesis protein. These preliminary results confirmed the potential of GWAS in identifying genetic variants present in KL13 and KL116 that could be associated with carbapenem resistance traits in Kp. The implementation of advanced methods, such as GWAS with increased antimicrobial resistance surveillance will potentially improve Kp infection treatment and patient outcomes.

Nutritional Properties of Milk from Dairy Ewes Fed with a Diet Containing Grape Pomace.

The aim of the present study was to evaluate the effects of a diet containing a 10% of grape pomace (GP) on the milk yield, chemical-nutritional characteristics, total phenolic compounds (TPCs), antioxidant activity (AOA), fatty acids and proteins profile of dairy ewe's milk. Forty-six ewes were dived into two groups: a control group (Ctrl), fed a standard diet, and an experimental group (GP+), whose diet was supplemented with 10% of GP on dry matter. The trial lasted 60 days and milk samples were collected and analyzed at the beginning (T0) and after 60 (T60) days. Dietary enrichment with GP did not affect the yield and the chemical composition of the milk. TPCs and AOA were not affected by the diet. After 60 days, the diet induced an increase in monounsaturated fatty acids (MUFA) and a decrease in medium chain saturated fatty acids (MCSFA), but the total saturated fatty acids (SFA), polyunsaturated fatty acids (PUFA), short chain saturated fatty acids (SCSFA) and long chain saturated fatty acids (LCSFA) were not modified. A decrease in the C14 desaturation index and an increase in the C18 index were also detected. Total caseins and whey protein were not affected by GP, even if a lower content of k-casein in GP+ milk compared to Ctrl milk was observed on the 60th day. The results of the present study suggest that 10% of GP can be included in the diet of lactating ewes without modifying milk gross composition but inducing significantly changes the fatty acid profile.

Angiostrongylus vasorum in foxes (Vulpes vulpes) and wolves (Canis lupus italicus) from Abruzzo region, Italy.

In Europe wildlife animals such as the red fox (Vulpes vulpes) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves (Canis lupus italicus) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection.

Physical, Nutritional, and Sensory Properties of Cheese Obtained from Goats Fed a Dietary Supplementation with Olive Leaves.

The aim of this study is to evaluate the physical, nutritional, and sensory properties of cheese obtained from goats fed a dietary supplementation with olive leaves (OL). Thirty Saanen goats were randomly allocated into two groups of 15 goats each, (1) a control group fed with a standard diet (CG), and (2) an experimental group (EG) fed an OL-enriched diet. The trial lasted for 30 days. The milk of each group was then collected and used to produce Caciotta cheese, which was analyzed at the beginning and at the end of the ripening period (60 days). The results showed a positive effect of dietary OL supplementation in improving the fatty acid profiles due to the significant increase of unsaturated fatty acids, mostly α-linolenic acid (C18:3 n-3), with the consequent reduction of the ω-6/ω-3 ratio, a condition commonly associated with an increased health functionality of food products. Moreover, improved oxidative stability was observed in cheese during ripening, a presumable consequence of the transfer into the milk of dietary bioactive compounds, mainly polyphenols of high biological value, and credited as a marked antioxidant potential. Furthermore, reduced lipolytic action was observed in 60-day ripened cheese, even if no significant changes in sensory properties were evidenced.

Application of the Micro Biological Survey analytical method for the determination of bacterial load in cow raw milk.

The aim of this study was to evaluate the performance of "Micro Biological Survey - MBS Test" in the enumeration of bacterial load in cow raw milk. The MBS test is based on a colorimetric method recently developed and patented by "Roma Tre" University, Italy. The evaluation of the performance of the MBS method was carried out by comparison with plate count at 30°C (gold standard) and flow cytometry. Thirteen independent set of experiments were performed analyzing a total of 104 samples of cow raw milk with the selected methods. Results obtained using the MBS method are comparable with those obtained with the plate count method at 30°C (CFU/mL) and flow cytometry technology; in particular, the results obtained with the MBS method are very close to plate count's at 30°C. On the other hand, there are statistically significant differences between these two methods' and flow cytometry technology's results that could be due to the different experimental conditions.

Angiostrongylus vasorum in 20 dogs in the province of Chieti, Italy.

After a case of Angiostrongylus vasorum (canine lungworm) was diagnosed in the province of Chieti (Italy) in early 2008, parasitological research was conducted to investigate the presence of the parasite in dogs in the area. A total of 178 dogs, 56 carcasses and 122 stool samples were examined between January and September 2008. The carcasses were examined for the presence of adult parasites in the right ventricle and pulmonary artery, and larval forms in the internal organ and brain tissues. The faeces were inspected for larval form L1 using three diagnostic methods that are currently used to test for endoparasites and larvae of bronchopulmonary strongyles. A total of 20 cases of canine angiostrongylosis were diagnosed (8.9%), with adult parasites being identified in 5 dogs, and L1 larvae in another 15. The anatomo-pathological examination of the carcasses of the dogs infested with adult nematodes revealed pneumonia, pleurisy, reddish foam in the trachea, effusion of serohaemorrhagic fluid in the thoracic cavity and enlarged mediastinic and mesenteric lymph nodes. Histological examination of the tissues showed serious, similar syndromes with lesions caused by colonisation of the kidneys, lymph nodes and brain by the parasites. Given the large number of cases confirmed in relation to the period of study (9 months), it is essential to include angiostrongylosis among the differential diagnoses made in clinical and post-mortem examinations of dogs in the province of Chieti and in the neighbouring areas.