Isolation and characterization of novel Staphylococcus aureus bacteriophage Hesat from dairy origin.

A novel temperate phage, named Hesat, was isolated by the incubation of a dairy strain of Staphylococcus aureus belonging to spa-type t127 with either bovine or ovine milk. Hesat represents a new species of temperate phage within the Phietavirus genus of the Azeredovirinae subfamily. Its genome has a length of 43,129 bp and a GC content of 35.11% and contains 75 predicted ORFs, some of which linked to virulence. This includes (i) a pathogenicity island (SaPln2), homologous to the type II toxin-antitoxin system PemK/MazF family toxin; (ii) a DUF3113 protein (gp30) that is putatively involved in the derepression of the global repressor Stl; and (iii) a cluster coding for a PVL. Genomic analysis of the host strain indicates Hesat is a resident prophage. Interestingly, its induction was obtained by exposing the bacterium to milk, while the conventional mitomycin C-based approach failed. The host range of phage Hesat appears to be broad, as it was able to lyse 24 out of 30 tested S. aureus isolates. Furthermore, when tested at high titer (108 PFU/ml), Hesat phage was also able to lyse a Staphylococcus muscae isolate, a coagulase-negative staphylococcal strain. KEY POINTS: • A new phage species was isolated from a Staphylococcus aureus bovine strain. • Pathogenicity island and PVL genes are encoded within phage genome. • The phage is active against most of S. aureus strains from both animal and human origins.

Immunity transfer in mule foals fed with good IgG quality colostrum.

While the passive transfer of immunity in horse and donkey foals has been extensively studied, there is limited information for mule foals. Immunoglobulin type G (IgG) and serum total protein concentration (TP) were assessed at different sampling times to evaluate the correlation between serum radial immunodiffusion (SRID) with electrophoresis, refractometry, and dry chemistry analyzer (Biuret), and to estimate serum IgG concentrations using serum TP in mule foals. We analyzed a total of 30 samples collected at birth, and at 6, 12, 24, and 48 h of life from 6 mule foals by SRID, electrophoresis TP, biuret TP, and refractometry TP. The SRID IgG concentration significantly increased from birth until T6 (p < 0.001). Serum TP analyzed with refractometry revealed differences between T0 and T12, T24 and T48 (p < 0.05), while a significant difference was observed with the biuret method between T0 and all the other sampling times (p < 0.001). A strong correlation was found between IgG SRID and biuret TP (r = 0.69, p < 0.001), and a good correlation existed between IgG SRID, refractometry TP, and electrophoresis TP (r = 0.44, p < 0.01 and r = 0.39, p < 0.05, respectively). All methods can be used to estimate the passive transfer of immunity in mule foals. TP refractometry and biuret TP values can be used to determine serum IgG concentrations in the blood of mule foals on their first day of life through the application of a specific equation.

Detour test performance of cloned minipigs from three different clone populations.

Genetics, the uterine environment, maternal behavior, and rearing conditions can all influence animal behavioral phenotypes. Some studies on cloned pigs have found no differences between the behavioral patterns of cloned and non-cloned animals. Other studies conducted on dogs have reported similarities in the behavior of cloned subjects. This study evaluated the performance of 12 cloned minipigs from three different clone populations (A, B, C) in a detour test around symmetric and asymmetric barriers. We measured the detour time and patterns, in order to investigate the pigs' cognitive abilities.The detour time and the detour entry/exit pattern were recorded. All the animals tended to keep a fixed entry/exit pattern instead of modifying it to accommodate changes in the working set. Significant differences in detour time were found among the populations, with animals belonging to population B being faster than the others, and also within each population.Our study is one of the few to assess the cognitive abilities of cloned minipigs. The results indicate that even animals belonging to the same cloned population may develop different cognitive, hence behavioral characteristics. Whether cloning can be utilized to obtain similar behavioral phenotypes therefore remains a matter of debate.

Linseed supplementation in the diet of fattening pigs: Effect on the fatty acid profile of different pork cuts.

Linseed supplementation improves the fatty acid (FA) profile of meat, primarily by increasing α-linolenic acid (ALA). Pork is one of the most consumed meats, though the high level of saturated FA, therefore, a change of the FA profile is needed to improve the health properties. The aim of this work was to study the influence of extruded linseed supplementation on the FA profile in 5 different pork cuts, increasing their nutraceutical properties. Sixty pigs were divided into two groups, which were fed two different diets: control (C) and experimental (L, 8% supplementation with extruded flaxseed). Five cuts were sampled: backfat (Bf), bacon (B), Boston shoulder (Bs), ham lean part (Hl) and ham fatty part (Hf). The L diet reduced fat content in Hf (-6%) and B (-11%), while the other cuts did not show differences. Moreover, L group showed a higher level of n-3 PUFA (approx. 9-fold), with a significant reduction in the n-6/n-3 from 20 to 2.5. In cuts rich in fat (Bf, B and Hf) of L group, the level of n-3 PUFA was higher than the thresholds set by EU to obtain the claim "Source of omega-3 fatty acids". In contrast, the lean cuts (Hl and Bs) did not exceed the threshold of n-3 PUFA required to obtain the claim, as consequence of the low level of fat. The results demonstrated that a diet with 8% extruded linseed enhanced the nutraceutical quality of pork meat.

Evaluation of Ultrasound Measurement of Subcutaneous Fat Thickness in Dairy Jennies during the Periparturient Period.

The body condition score (BCS) represents a practical but subjective method for assessing body fat reserves. Real time ultrasonography (RTU) has been proposed as an accurate method to objectively measure subcutaneous fat (SF) thickness and predict body fat reserves in cows, horses and donkeys. The aim of the present study was to describe RTU measures of SF thickness during periparturient period in jennies. The present prospective cohort study evaluated six dairy jennies. SF RTU were performed at 15 and 7 days before the presumptive delivery, and 2, 15 and 30 days after delivery. A portable ultrasound machine and multifrequency linear transducer (5-7.5 MHz) was used. RTU images were obtained in six sites (S1-S6). Results at each time point were reported as mean ± standard deviation and compared through time. A total of 180 images were evaluated. RTU technique was easy to perform and well tolerated. No statistically significant differences were found of each site during time, except for S2 and S6a: S2 at T2 and S6a at T1 were significatively different to values obtained at T5. The RTU mean values were above those reported by others, suggesting major physio-logical challenges related to energy balance and fat mobilization in pregnant jennies bred for milking production. BCS and sites through observational time have shown a good and reliable association. Our study could give preliminary indications on fat reserves in different body locations evaluated thanks to RTU and it show no significative variation of SF thickness, in pregnant and lactating jennies.

Body Condition Score Is Not Correlated to Gastric Ulcers in Non-Athlete Horses.

Equine Gastric Ulcer Syndrome (EGUS) is a worldwide disease of the stomach that can be found in different categories of horses. Different clinical signs may be present, but a large number of horses are asymptomatic. The aim of this study was to identify a possible correlation between body condition score (BCS) and EGUS in a population of horses. A total of 203 non-athlete horses were submitted for gastroscopies, and the presence and severity of lesions of the glandular and squamous mucosa were recorded. A board-certified veterinarian blinded to the gastroscopy results assessed the BCS of the horse. In the study population, no correlation was found between BCS and the presence of gastric lesions in either the glandular or the squamous mucosa. The disease of the squamous or glandular mucosa cannot be excluded based only on the presence of normal or increased BCS in non-athlete horses.

Validation of a new smart textiles biotechnology for heart rate variability monitoring in sheep.

Robust Animal-Based Measures (ABMs) are fundamental in order to assess animal welfare, however in semi-extensive sheep farming systems is not easy to collect ABMs without inducing additional stress in the animals. Heart rate variability (HRV) is a non-invasive technique of assessing stress levels related to animal welfare. It is considered a sensitive indicator of the functional regulatory characteristics of the autonomic nervous system. Several studies investigated the use of HRV for welfare assessment in dairy cows while research on sheep is scarce. Moreover, assessing HRV in small ruminants at pasture is critical because of the lack of a solution adoptable for field conditions. This study aimed to evaluate if a smart textiles technology is comparable to a Standard base-apex electrocardiogram (ECG) for measuring HRV in small ruminants. Eight healthy Massese dairy sheep were recruited. Standard base-apex ECG and smart textiles technology (Smartex ECG) were simultaneously acquired for 5 min in the standing, unsedated, unclipped sheep. The ECG tracings were recorded when animals were standing quietly. The Bland-Altman test and the linear regression analysis were applied after parameter extraction in time, frequency, and non-linear methods to compare Smartex against standard base-apex ECG systems. The Bland-Altman test was applied to all HRV extracted parameters (Mean RR, pNN50, RMSSD, LF/HF, SampEn, SD1, SD2, stdRR) to evaluate the agreement between the two different instruments, and a linear regression analysis was performed to evaluate the relationship between the two methods. The smart textiles biotechnology was simple to wear and clean. It can be worn without using glue and without shaving the sheep's wool, limiting animal handling and stress. Bland Altman test reported a robust agreement between the two systems. In fact, the regression analysis of HRV parameters showed that half of the parameters recorded had an R2 coefficient >0.75. Results also showed a very small reproducibility coefficient that indicated that the two methods were really close to each other. Smartex textiles technology can be used for HRV evaluation in sheep species as a potential ABM for animal welfare assessment.

Are Horse Owners Able to Estimate Their Animals' Body Condition Score and Cresty Neck Score?

Obesity and overconditioning are becoming more prevalent in horses, and are influencing the animal's health, activity, and reproductive efficiency. Body condition score (BCS) and cresty neck score (CNS) have been correlated with the development of various diseases. Many of the papers in the literature evaluate the prevalence of obesity and overconditioning in horse populations considering BCS given by owners. The aim of this study was to evaluate the validity of the owners' estimation of the degree of equine obesity or overconditioning by comparing BCS and CNS provided by each owner and a board-certified vet to a cohort of 259 horses and their agreement based on age, breed, sex, and activity of the animals. Overconditioning was present in 29% and 24% of horses according to the owners and the vet, respectively, and obesity in 2% and 1%, respectively. Agreement between BCS and CNS given by the owners and the vet was slight, with owners scoring horses either lower or higher than the vet. Agreement between the owners and the vet assessment was also influenced by age, breed, sex, and activity of the animals. Education of owners and caretakers to correctly assess BCS and CNS and regular evaluation of the animals throughout the year is important to prevent the development of obesity and overconditioning and related diseases.

Effect of a Phytogenic Feed Additive in Preventing Calves' Diarrhea.

The aims of the present study were to evaluate the preventive and the therapeutic effect of Stodi® as phytogenic feed additive rich in phenolic substances on the calf diarrhea, during the first 24 days of life. A total of 40 calves were included and randomly divided into Group C (control group) and Group T (treated group) with placebo or treatment administration started from the third day of life (T0). Calves belonged to group C received 2 L of warm water, while the calves assigned to group T received 2L of warm water plus 30 g of Stodi®. Solutions administration was maintained until day 21 (T21) that was the end of the experimental period. Calves were weighed at T0 and T21 to assess the average daily gain (ADG). Physical examination and fecal score evaluation were performed daily. The duration of a diarrheic episode, the age of the first diarrhea outbreak (TDE) and the frequency of diarrheic episodes were recorded. Complete blood count, methemoglobin and liver enzymes were evaluated at T0 and at T21 in all the calves by spectrophotometer and clinical chemistry analysis, respectively. Data were analyzed using a mixed model. A Chi-square and a Mann-Whitney test were also performed. No difference was found for ADG between the groups. The difference of mean age at TDE was not statistically significant between C and T group. The number of calves with diarrhea in the C group tended to be higher than that of T group (p = 0.13). Calves in group C spent more days with clinical sign of diarrhea compared to group T (p = 0.016). Complete blood count, methemoglobin and liver enzymes were within the reference ranges. The feed additive Stodi® seemed to be effective in shortening neonatal diarrhea episodes in calves thanks to the administration of 30 g per day of product. The fixed dosage of Stodi® used in our study did not show a preventive effect to reduce the incidence of calf diarrhea.

Differences of the Plasma Total Lipid Fraction from Pre-Foaling to Post-Foaling Period in Donkeys.

The period from late gestation to early lactation is characterized by changes in energy metabolism. The aim of the current study was to analyze the plasma total lipid fraction using gas chromatography (GC) analysis, in order to highlight metabolic changes from the pre-foaling to post-foaling periods. Eleven pluriparous dairy jennies (mean age of 11.88 ± 3.79 years) belonging to the Amiata donkey breed were enrolled. Blood sampling was performed at 15 days before foaling (T0), and 15 (T1), 30 (T2), 60 (T3), and 90 (T4) days after foaling, for biochemical and GC analysis. A total of 37 fatty acids were identified in plasma samples: 4 medium chain (MCFA), 24 long chain (LCFA), and 9 very-long chain (VLCFA) fatty acids. Among them, 20 fatty acids changed significantly, and two fatty acid showed a trend toward significance. Furthermore, the LCFA, saturated, unsaturated, monounsaturated, and polyunsaturated ω-3 fatty acids changed significantly during the study period. The main alterations were between T0 and the other time points and appeared to be related to lipid metabolism, cellular structure and function, and inflammatory and immune responses. Our findings reveal greater energy requirements at the end of gestation compared to early lactation in donkeys.

Designing Statistical Models for Holstein Rearing Heifers' Weight Estimation from Birth to 15 Months Old Using Body Measurements.

Body measurements could be used to estimate body weight (BW) with no need for a scale. The aim was to estimate heifers weight based on their body dimension characteristics. Twenty-five Holstein heifers represent the study group (SG); another 13 animals were evaluated as a validation group (VG). All the heifers were weighed (BW) and their wither height (WH), shin circumference (SC), heart girth circumference (HG), body length (BL), hip width (HW) and body condition score (BCS) were measured immediately after birth, and then weekly until 2 months and monthly until 15 months old. Equations were built with a stepwise regression in order to estimate the BW at each time using body measures for the SG. A linear regression was applied to evaluate the relationship between the estimated BW and the real BW. Equations found were to be statistically significant (r2 = 0.688 to 0.894; p < 0.0001). Three variables or fewer were needed for BW estimation a total of 11/23 times. Regression analysis indicated that the use of HG was promising in all the equations built for BW estimation. These models were feasible in the field; further studies will evaluate possible modifications to our equations based on different growing rate targets.

Standard measurement of the normal forelimb claw dimensions in Egyptian buffalo cows (Bubalus Bubalis) before and after functional trimming: a post-mortem study.

The aim of this study was to compare forelimb claw dimensions in buffalo cows, and to establish a reference values for functional claw trimming. Medial and lateral healthy front feet claws were evaluated from 20 young (YB) and 20 mature (MB) Egyptian buffalo cows (Bubalus bubalis). The dorsal wall length (TL), dorsal wall angle (TA), heel length (HBL), height (HBH) and width (HBW), sole (SL) and claw length (CL) and sole width (SW) were measured. Differences between YB and MB were evaluated using ANOVA test, while those between the lateral and medial claws with a paired t test. Before and after trimming, the HBL, HBW, HBH, SL and CL were significantly higher for both claws in MB, while the TA was significantly lower. Before trimming the lateral HBW and HBH were significantly higher than the medial, while the SL and CL of the medial claw were significantly higher than the lateral claw. After trimming, the lateral claw HBL was significantly higher in MB. The SW of the lateral claw was significantly higher than the medial claw in both YB and MB, and it did not change after trimming. These results can be considered as a guidance during front feet trimming in buffaloes.

Evaluation of Different Methods to Estimate the Transfer of Immunity in Donkey Foals Fed with Colostrum of Good IgG Quality: A Preliminary Study.

The aims of the present study were to evaluate the correlation between IgG Serum Radial Immunodiffusion (SRID), Electrophoresis Gamma Globulins (EGG), Electrophoresis Total Protein (ETP) and the serum total protein (TP) analyzed by refractometry and by a dry chemistry analyzer (Biuret) and to estimate serum IgG concentrations using serum TP. A total of 36 samples collected at four different times (birth, 6, 12, 24 h after birth) from nine Amiata donkey foals were evaluated with SRID, EGG, ETP, serum TP Biuret and refractometry. SRID IgG concentration increased significantly over time until T12. Serum TP analyzed with refractometry, electrophoresis and Biuret showed a statistically significant difference between T0 and T6 vs. T12 and T24. A good or strong correlation was found between different tests performed. Equations to quantify serum IgG were created and can be used for estimating the donkey foals' serum IgG in the first day of life. Serum TP refractometry showed a high correlation with SRID IgG (0.91) which may be a particularly useful and economic instrument to estimate the transfer of immunity in donkey foals during the first day of life. Further studies evaluating a high number of animals are needed in order to set specific cut-off values.

Presence and Characterization of Zoonotic Bacterial Pathogens in Wild Boar Hunting Dogs (Canis lupus familiaris) in Tuscany (Italy).

Domestic dogs (Canis lupus familiaris) used for wild boar (Sus scrofa) hunting may represent incidental hosts for several zoonotic pathogens. This investigation aimed to evaluate the presence of anti-Leptospira antibodies and the occurrence, antimicrobial resistance, and virulence of Salmonella spp., Yersinia enterocolitica, and Listeria monocytogenes in sera and rectal swabs collected from 42 domestic hunting dogs in the Tuscany region (Italy). Regarding Leptospira, 31 out of 42 serum samples (73.8%) were positive and serogroup Pomona was the most detected (71.4%) at titers between 1:100 and 1:400. Four Salmonella isolates (9.52%) were obtained, all belonging to serotype Infantis; two of them showed antimicrobial resistance to streptomycin, while pipB and sopE presence was assessed in all but one isolate. Concerning Yersinia enterocolitica, seven isolates (16.7%) were obtained, six belonging to biotype 1 and one to biotype 4. Resistance to amoxicillin-clavulanic acid, cephalothin, and ampicillin was detected. Biotype 4 presented three of the virulence genes searched (ystA, ystB, inv), while isolates of biotype 1 showed only one gene. No Listeria monocytogenes was isolated from dog rectal swabs. The results suggest that hunting dogs are exposed to different bacterial zoonotic agents, potentially linked to their work activity, and highlight the possible health risks for humans.

Evaluation of Brix Refractometry for the Estimation of Colostrum Quality in Jennies.

Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right half of nine Amiata jennies. A total of 72 colostrum samples were analyzed. A Friedman test with a Dunn's test for multiple comparisons was used for assessing the differences between the left and right half at each sampling time. No differences were found between the left and right halves; the average value was used to analyze the effect of sampling time on the IgG concentrations and Brix values. The relationship between colostrum IgG concentrations (SRID test) versus Brix value and Brix value versus time was analyzed using two different mixed linear models. A strong statistically significant relation has been found between IgG concentrations and Brix values (R2 = 0.84). The relation between IgG concentrations and Brix refractometer showed a cutoff point of 17% Brix score for the identification of high-quality colostrum. The Brix value (%) decreased continuously from 16.29 by 0.29 × hour. Jennies' and donkey foals' management may be greatly improved using this simple and cheap device.

Mammary Cistern Size during the Dry Period in Healthy Dairy Cows: A Preliminary Study for an Ultrasonographic Evaluation.

We evaluated the udder cistern (UC) size during the dry period using ultrasound. Forty healthy quarters were evaluated in both the longitudinal and cross-section of the UC. Quarters were evaluated at the drying-off (T0) and 24 h later (T1), then regularly until the end of the dry period (T7-T58), during the colostrum production phase (TCPP) and at 7 days in milking (T7PP). The Spearman test was applied to find the correlation between the ultrasonographic UC size (UUCS) assessment and time. The Friedman test and Dunn's test for multiple comparisons as a post-hoc test were performed to compare the forequarter and hindquarter cross-sections (FQCSs and HQCSs, respectively) and the forequarter and hindquarter longitudinal sections (FQLSs and HQLSs, respectively) at T0 vs. T58 vs. TCPP vs. T7PP. A total of 440 images were evaluated. A negative linear correlation between time and FQCS and FQLS (r = -0.95; p < 0.0004) and between time and HQCS and HQLS (r = -0.90; p < 0.002) was found. The UUCS decreased throughout the dry period, starting to increase at the beginning of the next lactation. Measuring the UUCS provides useful information for monitoring the dry period.

Evaluation of jennies' colostrum: IgG concentrations and absorption in the donkey foals. A preliminary study.

Immunoglobulin type G (IgG) concentration both in jennies' colostrum and in serum of donkey foals are mostly unknown in the first 24 h after delivery. The aims of the present study were to evaluate the IgG concentrations of colostrum during the first 24 h of lactation of Amiata jennies, the absorption of colostrum and the weekly body weight gain of the donkey foals. IgG concentrations were assessed in the jennies' colostrum and in the serum of donkey foals. Colostrum was collected in 9 jennies ready after delivery, and at 6, 12, 24 h after foaling from both halves. Serum was collected at the same sampling times from 9 donkey foals. Donkey foals were weighted at birth and then weekly until the 28th days of life. Temporal changes of IgG concentrations in dam's colostrum and in donkey foal serum were analyzed by a linear regression model and a general linear model, respectively. Results showed that colostrum IgG concentration were similar between the left and the right half. Colostrum IgG concentrations decreased continuously throughout the time in all jennies by 0.0244 Log10 mg/mL per hour. Serum IgG concentrations in donkey foals at birth was significantly lower compared to other times. No correlation was found between the colostrum IgG concentrations and the average weekly body weight gain of the donkey foal. The pattern of colostrum IgG levels in jennies and serum IgG concentration in donkey foals seem to be similar to what reported for equine. However, the donkey foals seem to be less agammaglobulinemic at birth compared to the horse foal. The pattern and both serum and colostrum concentrations evaluated in the Amiata donkeys were slightly different from results reported in other donkey breeds, underlying the importance of setting references specific to breed.

Pathotypes and Antimicrobial Susceptibility of Escherichia Coli Isolated from Wild Boar (Sus scrofa) in Tuscany.

Wild boar are among the most widespread wild mammals in Europe. Although this species can act as a reservoir for different pathogens, data about its role as a carrier of pathogenic and antimicrobial-resistant Escherichia coli are still scarce. The aim of this study was to evaluate the occurrence of antimicrobial-resistant and pathogenic Escherichia coli in wild boar in the Tuscany region of Italy. During the hunting season of 2018-2019, E. coli was isolated from 175 of 200 animals and subjected to antimicrobial resistance tests and PCR for detection of resistance and virulence factor genes. The highest resistance rates were against cephalothin (94.3%), amoxicillin-clavulanic acid (87.4%), ampicillin (68.6%), and tetracycline (44.6%). The most detected resistance genes were blaCMY-2 (54.3%), sul1 (38.9%), sul2 (30.9%), and tetG (24.6%). Concerning genes encoding virulence factors, 55 of 175 isolates (31.4%) were negative for all tested genes. The most detected genes were hlyA (47.4%), astA (29.1%), stx2 (24.6%), eaeA (17.1%), and stx1 (11.4%). E. coli was classified as Shiga toxin-producing E. coli (STEC) (21.7%), enterohemorrhagic E. coli (EHEC) (6.3%), enteroaggregative E. coli (EAEC) (5.1%), and atypical enteropathogenic E. coli (aEPEC) (3.4%). Enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and typical enteropathogenic E. coli (tEPEC) were not detected. Our results show that wild boars could carry pathogenic and antimicrobial-resistant E. coli, representing a possible reservoir of domestic animal and human pathogens.