Use of milk amyloid A in the diagnosis of subclinical mastitis in dairy ewes.

Subclinical mastitis (SM) is one of the most important diseases affecting dairy ewes worldwide, with negative impact on the animal health, farm income and public health. Animals with SM often remain untreated because the disease may not be revealed. Increase in somatic cell count (SCC) and positive bacteriology for mastitis pathogens in milk samples are indicative of SM but the evidence of only one of these alterations must suggest an uncertain SM (UM). UM is defined when positive bacteriological examination (Latent-SM) or SCC>500 000 cells/ml (non-specific-SM) are detected in milk. Nevertheless, SCC and bacteriological examination are expensive, time consuming and are not yet in use at the farm level in dairy ewes. Recently, a sensitive acute phase protein, amyloid A, displaying multiple isoforms in plasma and different body fluids including mammary secretion (milk amyloid A-MAA), has been investigated as a marker of mastitis in cows and, in a few studies, in sheep. The aim of this trial was to compare the concentration of MAA of single udder-halves in ewes with healthy udder-halves (HU-control group) and naturally occurring subclinical mastitis, both confirmed (SM group) and uncertain (UM groups: Latent-SM and non-specific-SM), for monitoring udder health. The reliability of a specific ELISA kit for the measurement of MAA was also tested. During a 3-month trial period, 153 udder halves were assigned to the experimental groups based on their health status: 25 with SM, 40 with UM (11 with latent-SM and 29 with non-specific-SM) and 88 HU. SCC and bacteriological analysis were performed to establish the control and subclinical mastitis groups. MAA concentrations in milk samples were measured using a specific commercially milk ELISA kit. The data were submitted to statistical analysis. Significant (P<0·05) differences among the groups SM, non-specific-SM and HU were detected with the SM having the highest level and HU the lowest. MAA concentration is affected by the udder health status and is a useful indicator of subclinical mastitis and increased SCC in sheep.

Brominated flame retardants (PBDEs and HBCDs) and perfluoroalkyl substances (PFASs) in wild boars (Sus scrofa) from Central Italy.

Twenty-six samples of wild boar liver and muscle from the Central Apennine Mountain (Italy) were analysed for 19 perfluoro-alkyl substances (PFASs), 10 polybrominated diphenylethers (PBDEs) and 3 hexabromocyclododecanes (HBCDs). All samples were analysed by gas chromatography-tandem mass spectrometry for PBDEs and liquid chromatography-tandem mass spectrometry for PFASs and HBCDs, using an in-house developed analytical procedure. The brominated flame retardants (BFR) levels in livers were negligible: Σ10PBDEs reached a maximum value of 0.079 µg/kg, whereas HBCDs were not quantified in almost all of the samples analysed. BFR concentrations in muscles were higher, but not significantly therefore, for Σ10PBDEs lower bound, a mean value of 0.045 µg/kg (0.005-0.155 µg/kg range) was measured, while α-HBCD was quantified with a maximum of 0.084 µg/kg in 9 of the samples. Only two muscles contained all 3 HBCD isomers at concentrations of approximately 0.200 µg/kg. Σ19PFAS in the 26 wild boar livers was in the range 31.9-228 µg/kg, with a mean value of 87.7 µg/kg, reaching levels significantly higher than in muscles, which exhibited a mean concentration of 3.08 µg/kg (0.59-9.12 µg/kg range). Perfluorooctanesulfonic acid (PFOS) was the most prevalent compound in all liver samples, accounting for more than half of the total PFASs contamination, confirming that the liver is the primary target organ for PFOS exposure Perfluorotridecanoic acid (PFTrDA), which accounts for 25-30-% of the total contamination, was the most abundant compound in the muscle, followed by PFOS. The estimated daily intake (EDIs) of BFRs remained below the estimated chronic human daily dietary intake (Dr,h) defined from European Food Safety Authority (EFSA). Furthermore, the exposure to PFASs in muscle was 7.7 times lower than the EFSA's tolerable daily intake (TDI). In contrast, exposure due to liver consumption was significant: the EDI exceeded the EFSA's 2020 TDI by approximately 7 times.

Health Status of the Eastern Grey Squirrel (Sciurus carolinensis) Population in Umbria: Results of the LIFE Project 'U-SAVEREDS'.

The introduction of the Eastern grey squirrel (Sciurus carolinensis) in Europe is one of the best-known cases of invasive alien species (IAS) colonisation, that poses a severe risk to the conservation of biodiversity. In 2003, it was released in a private wildlife park near the city of Perugia (Italy), where it is replacing the native Eurasian red squirrel (Sciurus vulgaris). The LIFE13 BIO/IT/000204 Project (U-SAVEREDS) was set up for the Sciurus vulgaris conservation in Umbria through an eradication campaign of grey squirrels. One hundred and fifty-four animals were analysed for bacteriological, mycological, virological, and serological investigations (C4 action). Sanitary screening showed that Sciurus carolinensis is a dermatophyte carrier, and therefore, it could cause public health issues for humans, considering its confident behaviour. Moreover, it has been marginally responsible for the spreading of Candida albicans, Coxiella burnetii, and Borrelia lusitaniae. Health status evaluation conducted on the Sciurus carolinensis population indicated that it is necessary to raise awareness of its impacts on biodiversity and human health. Moreover, the health status and behaviours of the IAS must be considered when control or eradication campaigns are planned.

Changes in plasma fatty acids profile in hyperketonemic ewes during early lactation: a preliminary study.

The transition from late pregnancy to early lactation is characterized by marked changes in energy balance of dairy ruminants. The mobilization of adipose tissue led to an increase in plasma non-esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB). The aim of this study was to analyze the total plasma fatty acids of healthy and hyperketonemic dairy ewes in early lactation through gas chromatography (GC) to evaluate metabolic alterations. An observational study was used with a cross-sectional experimental design. Forty-six Sarda dairy ewes were enrolled in the immediate post-partum (7 ± 3 days in milk) and divided into two groups according to serum BHB concentration: non-hyperketonemic group (n = 28; BHB < 0.86 mmol/L) and hyperketonemic group (n = 18; BHB ≥ 0.86 mmol/L). A two-way ANOVA included the effect of group and parity was used to evaluate differences in fatty acids (FA) concentrations. A total of 34 plasma FA was assessed using GC. 12 out of 34 FA showed a significant different between groups and 3 out of 34 were tended to significance. Only NEFA concentration and stearic acid were influenced by parity. The results may suggest possible links with lipid metabolism, inflammatory and immune responses in hyperketonemic group. In conclusion, GC represents a useful tool in the study of hyperketonemia and primiparous dairy ewes might show a greater risk to develop this condition.

Evaluation of the metabolomic profile through 1H-NMR spectroscopy in ewes affected by postpartum hyperketonemia.

Ketosis is one of the most important health problems in dairy sheep. The aim of this study was to evaluate the metabolic alterations in hyperketonemic (HYK) ewes. Forty-six adult Sardinian ewes were enrolled between 7 ± 3 days post-partum. Blood samples were collected from the jugular vein using Venosafe tubes containing clot activator from jugular vein after clinical examination. The concentration of ß-hydroxybutyrate (BHB) was determined in serum and used to divide ewes into assign ewes into: Non-HYK (serum BHB < 0.80 mmol/L) and HYK (serum BHB ≥ 0.80 mmol/L) groups. Animal data and biochemical parameters of groups were examined with one-way ANOVA, and metabolite differences were tested using a t-test. A robust principal component analysis model and a heatmap were used to highlight common trends among metabolites. Over-representation analysis was performed to investigate metabolic pathways potentially altered in connection with BHB alterations. The metabolomic analysis identified 54 metabolites with 14 different between groups. These metabolites indicate altered ruminal microbial populations and fermentations; an interruption of the tricarboxylic acid cycle; initial lack of glucogenic substrates; mobilization of body reserves; the potential alteration of electron transport chain; influence on urea synthesis; alteration of nervous system, inflammatory response, and immune cell function.

Comparison between Single- and Group-housed Pregnant Sows for Direct and Indirect Physiological, Reproductive, Welfare Indicators and Gene Expression Profiling.

Single- and group-housed pregnant sows were assessed during 8 weeks (4th and 8th to 14th) with regard to physiological, reproductive, welfare indicators and gene expression profiling. Compared to single-housed sows, group-housed sows had decreased non-return to estrus at 56 days after artificial insemination (AI) (83% vs 92%) and farrowing rate (78% vs 88%), respectively. Furthermore, group-housed sows showed a higher degree (p < 0.01) of advantageous physiological indicators, such as albumin (odds ratio [OR] = 4.4), alkaline phosphatase (OR = 1.5), bactericidal (OR = 3.2) and complement (OR = 24.3), and disadvantageous for alanine amino transferase (OR = 0.5), bilirubin (OR = 0.4), lysozyme (OR = 0.3) and C-reactive protein compared to single-housed. Eighty-seven genes related to immune response were underexpressed (log fold change ≤ 1.5; p < 0.05) during the 8th to 14th weeks in the group compared to single-housed sows, which in turn showed an immunomodulatory reduction on the expression of 43 genes during the 11th to 14th compared to the 4th week. Overall, the results were interpreted as indicative of greater comfort state of the group compared to single-housed sows.

Hepatitis E virus (HEV) genotype 3 diversity: Identification of a novel HEV subtype in wild boar in Central Italy.

In the last decade in Europe, the number of autochthonous cases of hepatitis E has significantly increased. Most of the cases arise from foodborne infections caused by the zoonotic hepatitis E virus (HEV) genotypes HEV-3 and HEV-4. Several human cases have been linked to consumption of raw or undercooked animal products of both pork (liver sausages) and wild boar meat. In this study, the occurrence of HEV infection was investigated in 611 livers and 88 paired lungs from wild boars collected during the hunting seasons of 2016-2020 in the Umbria-Marche Apennines (Central Italy). Using real-time reverse transcription polymerase chain reaction, 15 liver samples (2.45%) and one lung sample were found to be positive for HEV RNA. The phylogenetic tree built on the partial ORF2 gene revealed that the detected HEV strains belonged to HEV-3f (n = 5), HEV-3e (n = 1) and HEV-3c (n = 1) subtypes. Interestingly, 8 strains were genetically placed in a different cluster, further away from all other subtypes. To corroborate this finding, four complete genomes were obtained by next generation sequencing. The full genome of the HEV strains clustered together with another wild boar strain previously detected in Southern Italy in 2015 but the strains were divergent from all the HEV-3 strains classified in any subtype defined so far. Thus, these strains represent a novel subtype that might have originated in Italy, which we have tentatively named HEV-3n.

How the kinetic behavior of organic chickens affects productive performance and blood and meat oxidative status: a study of six poultry genotypes.

The aim of this study was to analyze the relationship between the kinetic behavior, carcass characteristics, oxidative status (blood and meat), and meat fatty acid profiles of 6 organically reared slower growing chicken genotypes (SrG). One hundred male chickens of 6 SrG were used: Ranger Classic (RC), Ranger Gold (RG), Rowan Ranger (RR), RedJA (RJ), CY Gen 5 JA87 (CY), and M22 × JA87 (M). Twenty chickens/genotype were selected to analyze behavior, while, 15 individuals were slaughtered and different traits were analyzed in the blood and drumstick meat. The variables were grouped into different principal components: kinetic activity (PC1, with explorative attitude as the highest score), productive performance (PC2, carcass and head/feet yields), blood (PC3, carbonyls, and TBARS) and meat (PC6, thiols, and TBARS) markers, technological traits (PC4, pH, and color), proximate meat composition (PCA5, moisture, lipids, protein, and ash), fatty acid profile, and nutritional indexes (PC7, IP, and PUFAn-3). Uni- and bivariate analyses showed a strong positive association between kinetic behaviors and blood and meat oxidation and a medium positive association with fatty acid profile and nutritional indexes, whereas a negative association was found between productive performance loads and the technological traits of meat. Generalized linear models showed that all PCs were influenced by genotype. In particular, CY and M resulted as less active genotypes; conversely, RR showed more kinetic activity, whereas RJ, RG, and RC exhibited intermediate levels of activity. Cluster analysis of kinetic behavior and blood or meat oxidative status highlighted 2 groups: nonwalking (NW: CY and M) and walking (W: RC, RG, RR, and RJ) animals. However, in the W group, another was visualized, constituted by genotypes with high kinetic activity resulting in the worst oxidative balance (Walking not trained-genotypes, Wnt: RR and RJ). The present results confirmed that the kinetic behavior of SrG genotypes is negatively correlated with productive performance. Furthermore, a significant association between kinetic behavior and blood (positively correlated) or meat (negatively correlated) oxidative status was noted. Such differences are mainly due to the intrinsic response of the genotypes used (i.e., training-walking capacity).

Milk Fatty Acids Composition Changes According to ß-Hydroxybutyrate Concentrations in Ewes during Early Lactation.

Ketosis is a metabolic disease of pregnant and lactating ewes linked to a negative energy balance which can cause different economic losses. The aims of this study were to understand the metabolic status of the early-lactating ewes and to identify biomarkers for early diagnosis of subclinical ketosis. Forty-six Sarda ewes were selected in the immediate post-partum for the collection of the biological samples. A blood sample from the jugular vein was used to determine ß-Hydroxybutyrate (BHB) concentrations. Animals were divided into two groups: BHB 0 or healthy group (n = 28) with BHB concentration < 0.86 mmol/L; and BHB 1 or subclinical ketosis (n = 18) with a BHB concentration ≥ 0.86 mmol/L. Ten mL of pool milk were collected at the morning milking for the analyses. The concentration of 34 milk fatty acids was evaluated using gas chromatography. Two biochemical parameters and 11 milk fatty acids of the total lipid fraction presented a p-value ≤ 0.05. The study revealed different relationships with tricarboxylic acid cycle, blood flows, immune and nervous systems, cell functions, inflammatory response, and oxidative stress status. Eight parameters were significant for the receiver operating characteristic (ROC) analysis with an area under the curve greater than 0.70.

Natural immunity in conventionally and organically reared turkeys and its relation with antimicrobial resistance.

Suboptimal animal welfare may affect natural immunity, rendering animals more susceptible to environmentally conditioned diseases, including those requiring antimicrobial treatment, which may promote antimicrobial resistance (AMR) in bacterial populations. Herewith, we tested the hypothesis that conventionally raised turkeys have higher levels of AMR in indicator Escherichia coli bacteria, but lower levels of natural immunity, as compared to turkeys reared under organic conditions. Litter and serum samples were collected from 28 conventional and 4 organic turkey farms: E. coli isolates from litter were tested for resistance to 14 antimicrobials, while 3 parameters of natural immunity (i.e., lysozyme, hemolytic complement levels, and serum bactericidal activity) were assessed in the sera. Resistant E. coli isolates were identified in both conventional and organic farms but generally more frequently in conventional farms. High rates of resistance to ampicillin (96%), tetracycline (95%), streptomycin (82%), sulfamethoxazole (80%), ciprofloxacin (73%), and trimethoprim (71%), as well as high rates of multiresistance, were observed in conventional farms. Organically raised turkeys had significantly higher levels of lysozyme and serum bactericidal activity than conventional turkeys, and these levels were also higher in turkeys housed in farms where AMR frequency was lower. Findings support the hypothesis that conventional farming conditions may affect turkeys' natural immunity, rendering the animals more susceptible to environmentally conditioned diseases requiring antimicrobial treatment, which would in turn promote AMR. Reducing AMR in turkey farming is therefore more likely to be successful when considering animal welfare as an option to reduce the need of antimicrobial use.

Plasmatic Profile of Pregnancy-Associated Glycoprotein (PAG) during Gestation and Postpartum in Sarda and Lacaune Sheep Determined with Two Radioimmunoassay Systems.

This study was carried out to determine ovine pregnancy-associated glycoprotein (oPAG) levels in the plasma of Sarda and Lacaune ewes throughout gestation and in the first month postpartum, using two heterologous radioimmunoassays (RIA-706 and RIA-srPool) and to study the correlations between PAG levels and fetal gender and number. On Day 18 of pregnancy, PAG concentrations were detected in 90.1% and 80.8% of Sarda pregnant ewes with RIA-706 and RIA-srPool, respectively; and in 90% and in 75% of Lacaune pregnant ewes with RIA-706 and RIA-srPool, respectively. From Day 30, PAG concentrations were detected in all pregnant ewes by using both RIA methods. In the postpartum period, the PAG concentrations in Sarda ewes decreased rapidly reaching minimal levels (<1 ng/mL) on day 28 using both RIA-706 and RIA-srPool. In Lacaune ewes, PAG-706 levels were higher than PAG-srPool from parturition until the last day of observation (Day 28 postpartum). It was also observed that mean concentrations were higher in multiple than in single pregnancies in Sarda and Lacaune ewes. Moreover, due to the rapid disappearance of PAG concentration following lambing, a cut-off limit in postpartum animals was not required as a means for detecting a new pregnancy.

Welfare Assessment in Shelter Dogs by Using Physiological and Immunological Parameters.

This study aimed to evaluate the state of welfare of a group of dogs during the first month after entering the shelter by using different stress parameters. Blood and fecal samples were collected from a group of 71 dogs at the time of admission to the shelter. In 46 of these dogs, sampling was repeated after four weeks. Well-recognized welfare biomarkers, such as fecal cortisol and leukocytes, as well as some innovative parameters (ß-endorphin and lysozyme) were determined. Uni- and multivariate statistical analyses were used to evaluate their interactions and changes over time. Neutrophils (p < 0.01), lysozyme (p < 0.05), and fecal cortisol (p < 0.05) decreased, while lymphocytes (p < 0.05) increased after four weeks compared to the first days of being in the shelter, suggesting an improvement in the dogs' welfare over time. A principal component analysis extracted three bipolar components (PCs), explaining 75% of the variance and indicating negative associations between neutrophil and lymphocyte (PC1), lysozyme and ß-endorphin (PC2), cortisol and lysozyme (PC3). The associations between these variables within each PC also confirmed the intricate relationships between the hypothalamic-pituitary-adrenal (HPA) axis and the immune system as well as the importance of a multiparametric approach in evaluating welfare.

Benefits of dietary supplements on the physical fitness of German Shepherd dogs during a drug detection training course.

A high standard of physical fitness is an essential characteristic of drug detection dogs because it affects not only their ability to sustain high activity levels but also their attention and olfaction efficiency. Nutritional supplements could improve physical fitness by modulating energy metabolism, oxidative processes, and perceived fatigue. The aim of this study was to investigate the physiological and biochemical changes induced by submaximal exercise on drug detection dogs (German Shepherd breed) and to assess whether a dietary supplement improves their physical fitness. During a drug detection dog training course, seven dogs were fed with a basal diet (Control Group) for three-month period, while a further seven dogs were fed with a basal diet as well as a daily nutritional supplement containing branched-chain and limiting amino acids, carnitine, vitamins, and octacosanol (Treatment Group). At the end of this period, individual physical fitness was assessed by making each subject take a graded treadmill exercise test. A human heart rate monitor system was used to record the dog's heart rate (HR) during the treadmill exercise and the subsequent recovery period. The parameters related to HR were analysed using nonparametric statistics. Blood samples were collected before starting the nutritional supplement treatment, before and after the treadmill exercise and following recovery. Linear mixed models were used. The dietary supplements accelerated HR recovery, as demonstrated by the lower HR after recovery (P<0.05) and Time constants of HR decay (P<0.05), and by the higher Absolute HR Recovered (P<0.05) recorded in the Treatment group compared with the Control dogs. The supplemented dogs showed the lowest concentrations of creatine kinase (CK; P<0.001), aspartate aminotransferase (AST, P<0.05) and non-esterified fatty acids (NEFA; P<0.01) suggesting a reduction in muscle damage and improvement of energy metabolism. These data suggest that this combined supplement can significantly enhance the physical fitness of drug detection dogs.

Housing Rabbit Does in a Combi System with Removable Walls: Effect on Behaviour and Reproductive Performance.

We evaluated the effects of two types of colony cages, in which rabbit does were always in a group (C1), and where they were in combi cages furnished with removable internal walls to allow both individual and grouphousing (C2), in addition to the control group (C: conventional individual cage), on welfare, reproductive performance, and global efficiency. Forty-eight New Zealand White nulliparous rabbit does underwent artificially insemination (AI) and were divided into three groups, and reared in the different systems for about 1 year. The reproductive rhythm provides AIs at weaning (30d). In the C1 system, does were continuously grouped, while in C2, walls were inserted four days before kindling and removed 1week after it (60% of the timesheet in group). Reproductive traits and behaviour were evaluated during the entire year. The behavioural observations were performed around days 7, 36, and 44, corresponding to the inclusion of the does in the maternal cages, the insertion of walls four days before kindling, and the removal of the walls 1week after parturition in the C2 group, respectively. The percentages of does with severe skin injuries and the distribution of the injuries on different parts of body were also registered. Does reared in conventional cages showed the greatest presence of stereotype behaviours, while the C1 group showed the highest (p < 0.05) incidence of aggressiveness after regrouping (attack, dominance features, and lower allo-grooming) in comparison to the C2 group (17% and 22%, in C2 and C1 does, respectively).Individually caged does achieved the best productive performance (sexual receptivity, fertility, kindling rate, and number of kits born alive and at weaning). The C1 group showed the lowest performance (p < 0.05), whereas C2 showed an intermediate one. Does housed in the combi cage (C2) had higher (p < 0.05) receptivity and fertility rates and higher numbers of kits born alive and at weaning (79.2% and 76.2%; 7.95 and 7.20, respectively) than the C1 group, but lower values (p < 0.05) than does that were individually housed.

Dehydrated Alfalfa and Fresh Grass Supply in Young Rabbits: Effect on Performance and Caecal Microbiota Biodiversity.

The improvement of rabbit gut microbiota by modifying nutritional components of the feed or favoring its early intake of feed has been previously investigated. The early administration of dehydrated alfalfa (A) or fresh grass (G) for rabbits, during the peri-weaning period (30 and 45 days of age), and their effect on performance and caecal microbiota compared to a standard diet (C) were evaluated. Until 15 days of age, nine litters/group were housed in the maternal cage and milked once per day. From 15 to 30 days, the young rabbits could consume both milk and solid feed (pelleted for C or supplemental feed for A and G). At 30 days of age, the rabbits were weaned and, until 45 days, were kept in single cages following the same dietary protocol. No significant changes were found in the milk intake or the individual weight of young rabbits at 30 and 45 days. The caecal Firmicutes/Bacteroidetes (bacterial phyla ratio) increased with age (from 2.43 to 6.05 on average, at 30 and 45 days). The Ruminococcaceae/Lachinospiraceae (bacterial family ratio) was highest in the A group at both ages, followed by G then C. The early administration of dehydrated alfalfa is a promising solution to improve health status by favoring an appropriate digestive microbiota.

Dietary Saccharomyces cerevisiae boulardii CNCM I-1079 Positively Affects Performance and Intestinal Ecosystem in Broilers during a Campylobacter jejuni Infection.

In poultry production, probiotics have shown promise to limit campylobacteriosis at the farm level, the most commonly reported zoonosis in Europe. The aim of this trial was to evaluate the effects of Saccharomyces supplementation in Campylobacter jejuni challenged chickens on performance and intestinal ecosystem. A total of 156 day old male Ross 308 chicks were assigned to a basal control diet (C) or to a Saccharomyces cerevisiae boulardii CNCM I-1079 supplemented diet (S). All the birds were orally challenged with C. jejuni on day (d) 21. Live weight and growth performance were evaluated on days 1, 21, 28 and 40. The histology of intestinal mucosa was analyzed and the gut microbiota composition was assessed by 16S rRNA. Performance throughout the trial as well as villi length and crypt depth were positively influenced by yeast supplementation. A higher abundance of operational taxonomic units (OTUs) annotated as Lactobacillus reuteri and Faecalibacterium prausnitzii and a lower abundance of Campylobacter in fecal samples from S compared to the C group were reported. Supplementation with Saccharomyces cerevisiae boulardii can effectively modulate the intestinal ecosystem, leading to a higher abundance of beneficial microorganisms and modifying the intestinal mucosa architecture, with a subsequent improvement of the broilers' growth performance.

Reference values for serum amyloid A, haptoglobin, lysozyme, zinc and iron in healthy lactating Lacaune sheep.

Acute-phase proteins and trace elements are considered biomarkers of the immune response to infection, inflammation, trauma and other pathological conditions, as well as indicators of the health status and productivity of farm animals. Given the scarcity of published data on this topic, the purpose of this study was to determine the serum levels of serum amyloid A (SAA) and haptoglobin, as well as lysozyme, iron and zinc in clinically healthy Lacaune sheep in lactation months 3-7 ranging in age from 2 to 6 years. The mean serum levels for SAA (12.2 µg/mL), lysozyme (1.47 µg/mL), zinc (78.9 µg/dL) and iron (26.6 mmol/L) differed from those found in other sheep breeds and indicate considerable variations depending on breed, age and physiological status of the sheep. Reference values for clinically healthy mid-lactating Lacaune sheep were determined by using well-described and modern analytical and statistical methods. The reference intervals may be used to determine the health and welfare of lactating Lacaune sheep and may serve as a starting point to investigate diseases.

Retrospective Biomolecular Investigation of Coxiella burnetii and Leptospira spp. DNA in Cases of Abortion, Stillbirth and Neonatal Mortality in Dogs and Cats.

Abortion and neonatal mortality are events that can occur in breeding bitches and queens. It has been reported that up to 55% and 33% of these cases remain without a known cause, respectively, in canine and feline pregnancies. Unusual abortigenic and potentially zoonotic agents, including Coxiella burnetii and Leptospira spp., may be involved in these cases. C. burnetii is able to cause reproductive disorders in cattle, sheep and goats, and cases of abortion have been observed in dogs and cats. Moreover, several outbreaks of C. burnetii infection in humans have been caused by delivering bitches and queens, and some of these animals experienced abortion. Leptospira interrogans sensu lato is able to cause abortion or stillbirth in several animal species and its abortigenic role has occasionally been described in bitches and queens. The aim of this study was to search for C. burnetii and Leptospira spp. DNA in a retrospective series of 103 cases of canine and feline abortion, stillbirth, and neonatal mortality submitted for the identification of possible infectious agents. One hundred and fifty-one specimens were tested using PCR assays and found negative for C. burnetii and Leptospira DNA. However, in 49 samples (47.6%) other infectious causes of abortion, stillbirth, and neonatal mortality were identified. These results showed that C. burnetii and Leptospira spp. are probably not common abortigenic agents or causes of neonatal deaths in dogs. However, given the potential abortigentic and zoonotic role of these agents, surveillance of canine and feline abortion, stillbirth, and neonatal mortality could be advisable for a systematic investigation of these events.

Circulating miRNAs as Putative Biomarkers of Exercise Adaptation in Endurance Horses.

Endurance exercise induces metabolic adaptations and has recently been reported associated with the modulation of a particular class of small noncoding RNAs, microRNAs, that act as post-transcriptional regulators of gene expression. Released into body fluids, they termed circulating miRNAs, and they have been recognized as more effective and accurate biomarkers than classical serum markers. This study examined serum profile of miRNAs through massive parallel sequencing in response to prolonged endurance exercise in samples obtained from four competitive Arabian horses before and 2 h after the end of competition. MicroRNA identification, differential gene expression (DGE) analysis and a protein-protein interaction (PPI) network showing significantly enriched pathways of target gene clusters, were assessed and explored. Our results show modulation of more than 100 miRNAs probably arising from tissues involved in exercise responses and indicating the modulation of correlated processes as muscle remodeling, immune and inflammatory responses. Circulating miRNA high-throughput sequencing is a promising approach for sports medicine for the discovery of putative biomarkers for predicting risks related to prolonged activity and monitoring metabolic adaptations.

Protective effects of platelet-rich plasma against lidocaine cytotoxicity on canine articular chondrocytes.

BACKGROUND: Lidocaine (LD) is one of the most commonly used local anesthetics for performing arthroscopic surgery and managing of osteoarthritic pain in both human and veterinary medicine. However, over the last years, several studies have focused on the chondrotoxic effects of LD. In order to ensure that intra-articular lidocaine is safe to use, treatments aimed at mitigating chondrocyte death have recently been investigated. The aim of this study is to evaluate the possible protective effects of platelet-rich plasma (PRP) against LD cytotoxicity on canine articular chondrocytes. RESULTS: Articular canine chondrocytes, were exposed to 1% or 1.8% LD alone or in co-presence with 10% PRP for 30 min. In order to evaluate the effects of PRP pre-treatments, experiments were carried out on cells cultured in serum-free medium-or in medium supplemented with 10% PRP or 10% fetal bovine serum. Cell viability was evaluated by methyl thiazolyl tetrazolium assay and cell apoptosis was analyzed by flow cytometry using annexin V-fluorescein isothiocyanate/propidium iodide. The results showed that LD significantly reduced canine chondrocytes viability, probably due to apoptosis induction. Pre-treatment or the co-presence of PRP in the media restored the number of viable chondrocytes. The PRP also seemed to protect the cells from LD-induced apoptosis. CONCLUSIONS: Pre-treatments and/or the simultaneous administration of PRP reduced LD-induced cytotoxicity in canine chondrocytes. Further in vivo studies are required to determine whether PRP can be used as a save protective treatment for dogs receiving intra-articular LD injections.