Refinement and revalidation of the Equine Ophthalmic Pain Scale: R-EOPS a new scale for ocular pain assessment in horses.

This study addresses the refinement and revalidation of a composite pain scale that focuses on equine facial expressions and behavioural indicators as exhibitions of ophthalmic pain. This scale included only Behavioural and Facial and Ocular expression indicators and, compared to the first version of Equine Ophthalmic Pain Scale (EOPS), item descriptors and related ratings were changed. Thirteen horses with ocular diseases that required medical or surgical treatment were enroled (group P). In each animal, the refined EOPS (R-EOPS) was applied prior to any treatment (T0) and one week later (T7). The R-EOPS was applied twice, 7 days apart, to 16 healthy control horses (group C). Two 30-second videos were recorded each time to allow the retrospective analysis by eight observers. Inter-observer reliability of items was moderate or substantial (Krippendorff's alpha, Kα>0.40) while their intra-observer reliability was substantial or almost perfect for most items (Kα ≥0.61). Both inter- and intra-observer reliability of Total Score (TS) were however excellent (Intraclass Correlation Coefficients, ICC>0.75). The TS also showed good reproducibility (Kendall coefficient=0.786, ICC=0.684) and high consistency of its items (Cronbach's α=0.847). The comparison between groups as well as the sensitivity and specificity values supported the validity of the R-EOPS. In particular, for each extra point added to the TS, the risk of the horse having pain increased by more than two times (Odds Ratio=2.079, 95%CI=1.542-2.804; P<0.001). The Receiver Operating Characteristic analysis identified 6 as the threshold value of R-EOPS for discriminating horses with ocular pathology (sensitivity=83%, specificity=100%). This scale may be an effective tool for reliably assessing the pain level in horses with ophthalmic diseases and potentially guiding pain management although it still requires large-scale application and external validation.

Gene expression of pregnancy-associated glycoproteins-1 (PAG-1), interferon-tau (IFNt) and interferon stimulated genes (ISGs) as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows.

The aim of this study was to determine the presence of Pregnancy-associated glycoprotein -1 (PAG-1) mRNA expression in the maternal circulation of pregnant buffaloes during the early stage of pregnancy. Contemporaneously, the mRNA expression levels of Interferon-tau (IFNt) and some Interferon stimulated genes (ISGs) (interferon stimulated gene 15 ubiquitin-like modifier interferon, ISG15; Mixoviruses resistance 1 and 2, MX1 and MX2; 2',5'-oligoadenylate synthase 1,OAS1) were evaluated in order to expand our knowledge of the molecular processes involved in the early stages of pregnancy and to identify potential biomarkers of maternal-fetal cellular interaction in buffalo. The study was conducted on 38 synchronized and artificially inseminated buffalo cows (d 0), divided ex post into 3 groups: Pregnant (n = 17), Non-pregnant (n = 15) and Embryo mortality (n = 6). Blood samples were collected on d 14, 19, 28 and 40 after artificial insemination (AI) for peripheral blood mononuclear cells (PBMCs) isolation. Expression levels of mRNA of PAG-1, IFNt, ISG15. MX1, MX2 and OAS1 were measured using RT-qPCR. No significant changes were observed in IFNt and PAG gene expressions between groups, while significant differences (p < 0.001) were found for ISG15, MX1, MX2, and OAS1. Pairwise comparisons revealed that the differences between groups occurred on days 19 and 28 post-AI. ISG15 proved to have the best diagnostic performance for distinguishing between pregnant animals and animals that experienced embryo mortality with the ROC analysis. According to the results of the univariate analyses, day 19 was identified as the most indicative to discriminate between groups while the most reliable genes for this differentiation were ISG15, MX1 and MX2. MX2 proved to be the best gene for discriminating pregnant buffaloes using the discriminant analysis, while MX1 was the gene that best predicted embryo mortality. Our results showed that among PAG-1, IFNt and ISGs expression as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows, ISGs proved to be the best peripheral biomarkers for predicting pregnancy and embryonic mortality during the peri-implantation period. These insights into the mechanisms behind maternal-fetal interaction and the development of a method for the early detection of embryo distress may enable us to implement effective strategies to support embryo survival.

Development and preliminary validation of a pain scale for ophthalmic pain in horses: The Equine Ophthalmic Pain Scale (EOPS).

The purpose of this study was to describe the development and preliminary validation of a composite pain scale, called the Equine Ophthalmic Pain Scale (EOPS), to assess ocular pain in horses. Indicators associated with ocular pain were selected and classified as behavioural, physiological or ocular expressions. Eight horses diagnosed with ocular or adnexa diseases that required medical or surgical treatment were enrolled in the study (group P). The developed EOPS was applied at the baseline (T0) and 1 week later (T7). Moreover, the EOPS was applied twice, 1 week apart, to 15 healthy control horses (group C). Videos of 60-80 s duration of all assessments were retrospectively analysed by seven masked observers, who scored items included in the behavioural and ocular expression categories of the EOPS. The inter- and intra-observer reliability was excellent (intraclass correlation coefficients ≥0.75) for most of the scored items. Cronbach's alpha (0.76) indicated that the EOPS had good internal consistency. The total score (TS), calculated as the sum of all scores, differed between groups C and P at T0 (P < 0.001) and reduced after medical/surgical treatment in group P (P = 0.017), indicating the responsiveness of the EOPS. Moreover, the area under the curve (AUC=0.918, 95% confidence interval = 0.815-1.000; P < 0.001) indicated that the EOPS was very accurate for distinguishing healthy from pathological animals. Sensitivity and specificity of EOPS to identify horses with ocular pathology (at the optimal cut-off, i.e. TS ≥ 7) were 81.3% and 100.0%, respectively. However, 'overall behaviour', 'position inside the box', 'ear movements' and 'head position' items as well as physiological parameters, showed sub-optimal reliability, consistency and/or item-total correlation, suggesting that there is still room to improve this composite scale.

Fatty acid profile, oxidative status, and content of volatile organic compounds in raw and cooked meat of different chicken strains.

Chicken meat is rich in unsaturated fatty acids. Therefore, it is more susceptible to lipid oxidation and production of volatile organic compounds (VOC). In this study, we evaluated the fatty acids, antioxidants, and VOC profiles of raw and cooked meat samples derived from 4 strains of chicken differing in their growth rates, which were as follows: slow-growing (SG, Leghorn), medium-growing (MG, Hubbard and Naked Neck), and fast-growing (FG, Ross). The VOC profile of meat was measured using proton-transfer reaction-mass spectrometry (PTR-MS). The VOC were identified using PTR-time of flight-MS (PTR-ToF-MS). The data were analyzed using both univariate and multivariate models. Twenty main VOC were identified, which were classified into the following chemical categories: aldehydes, alkadienes, alkenes, furans, amides, alcohols, and other compounds. Our results revealed that the chicken genotype and the method of cooking strongly influenced the VOC profile of the meat. Identifying the relationships between these traits allowed us to highlight the trade-off of the main substrates such as n-3 and n-6 polyunsaturated fatty acids (PUFA), protective substances (antioxidants), and degradation products (VOC) of the poultry meat produced during cooking. The extent of VOC production and n-3 loss was found to be higher for the SG genotype. Reduction of n-6 was higher in MG, whereas small losses in antioxidants and PUFA were observed in the FG genotype, consequently, resulting in the lowest production of VOC. The SG and MG are genotypes more active from a kinetic point of view respect to the FG ones. For this reason, in the FG genotypes, the antioxidants are less involved in the oxidative stress induced by the movement; thus, they were available to protect the lipid of the meat during the cooking process. These results suggested that the use of SG and MG genotypes requires a specific dietary protocol (i.e., increasing the antioxidants content) to counteract the lipid oxidations in all the phases: in vivo, postmortem, and during/after cooking.

A cross-talk between blood-cell neuroplasticity-related genes and environmental enrichment in working dogs.

This study aims to identify a panel of blood-cell neuroplasticity-related genes expressed following environmental enrichment stimulation (EE). The Drug detection (DD) training course was an excellent model for the study of EE in the working dog. This research is divided into two experimental trials. In the First Trial, we identified a panel of blood-cell neuroplasticity related-genes associated with DD ability acquired during the training course. In the Second Trial, we assessed the EE additional factor complementary feeding effect on blood-cell neuroplasticity gene expressions. In the First and Second Trials, at different time points of the DD test, blood samples were collected, and NGF, BDNF, VEGFA, IGF1, EGR1, NGFR, and ICE2 blood-cell neuroplasticity related-genes were analyzed. As noted in the First Trial, the DD test in working dogs induced the transient up-regulation of VEGFA, NGF, NGFR, BDNF, and IGF, immediately after the DD test, suggesting the existence of gene regulations. On the contrary, the Second Trial, with feeding implementation, showed an absence of mRNA up-regulation after the DD test. We suppose that complementary feeding alters the systemic metabolism, which, in turn, changes neuroplasticity-related gene blood-cell mRNA. These findings suggested that, in working dogs, there is a cross-talk between blood-cell neuroplasticity-related genes and environmental enrichment. These outcomes could be used to improve future treatments in sensory implementation.

Occurrence and antimicrobial susceptibility of Campylobacter spp. on fresh and refrigerated chicken meat products in Central Italy.

This study investigated the presence and the level of Campylobacter spp. contamination in 41 thigh samples (with skin) and 37 skinless breast samples collected at the end of slaughter (T1) and after 10 day period at refrigeration temperature (4°C) (T2), corresponding to their commercial shelf life. The isolates were phenotypically classified as Campylobacter spp. and successively identified by conventional multiplex PCR. The antimicrobial susceptibility of the isolates from fresh thigh and breast samples was also determined via the microdilution method (MIC) in Eucamp microtitre plates with known scalar concentrations of: gentamicin (GEN), streptomycin (ST), ciprofloxacin (CIP), tetracycline (TET), erythromycin (ERY), and nalidixic acid (NA). A greater percentage of positivity for Campylobacter spp. (P < 0.001) was observed in thighs and C.jejuni appeared to be the most common species identified at this level (P < 0.001) followed from its association with C.coli. There was a global reduction of Campylobacter spp. in both thigh and breast samples at T2 (P < 0.001) showing that the refrigeration was able to reduce Campylobacter count. The prevalence of resistance to CIP, TET, NA, and ERY was evidenced for C.jejuni and C. coli. The co (TET-NA, CIP-NA) and multiple resistant (CIP-TET-NA, CIP-TET- NA-ERY) isolates came from the thigh products. It should be highlighted the presence of Campylobacter spp. isolates resistant to ST occurred in breast samples, responsible for the ST-CIP co-resistance and ST-CIP-TE multi-resistance profiles, higher in breast than in thigh products (P > 0.001). The presence of Campylobacter isolates resistant to ST can be further investigated since it is used for therapeutic treatment of several bacterial diseases in humans.

The different effects of linseed and fish oil supplemented diets on insulin sensitivity of rabbit does during pregnancy.

This study investigates the effects of linseed (rich in ɑ-linolenic acid (ALA)) and fish oil (rich in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA)) supplementation on the insulin resistance of pregnant rabbits. Two months before insemination, the rabbits (15 animals/group) were fed different diets: commercial standard (group C), supplemented with 10% extruded linseed (group L), and 3% fish oil (group FO). The L group does showed both the highest feed intake before AI (P < 0.01) and the highest body weight (BW) throughout pregnancy (P < 0.001). The L does yielded less milk than the C does (P < 0.001); however, no differences were observed in either weight or size of litter at weaning. Regardless of diet, insulin concentrations and HOMA-IR values were higher during the first half of pregnancy (P < 0.001). Nevertheless, the L does showed higher mean insulin concentrations than FO rabbits (P < 0.01) and the lowest glucose clearance (P < 0.01) during pregnancy. On the other hand, pregnant FO rabbits showed the lowest glucose concentrations (P < 0.05) and the lowest Homeostasis model assessment values for insulin resistance (HOMA-IR, P < 0.05) as well as a faster restoration of baseline glucose levels following glucose load (P < 0.001). Before and during pregnancy, the BW of the rabbits was positively related to fasting sample- and tolerance test-derived indices of insulin resistance (P < 0.05) suggesting that a high pre-pregnancy BW predisposes to gestational insulin resistance. Linseed supplementation increased BW and predisposed to insulin resistance during pregnancy; whereas, fish oil improved insulin sensitivity without significant changes in BW.

Serum concentration dynamic of energy homeostasis hormones, leptin, insulin, thyroid hormones, and cortisol throughout canine pregnancy and lactation.

Pregnancy and lactation represent critical periods of canine reproductive life and different hormones are required to maintain homeostasis and the correct energy requirements. The aim of this study was to evaluate leptin, insulin, triiodotyronine (T3), thyroxin (T4) and cortisol serum concentrations to better describe the canine maternal energy metabolism. Twelve Bloodhound bitches that had at least one litter and were considered to be healthy submitted blood samples every 15 days starting from day 0 (ovulation) throughout pregnancy up to the end of lactation (day 120). We found that leptin concentrations increased from day 0 to reach maximum value at day 45 (P < 0.001), decreased at day 60 (P < 0.05) and rose again at day 105 and day 120 (P < 0.05 and P < 0.01 respectively). Insulin concentrations significantly increased during pregnancy peaking at day 60 (P < 0.05). After parturition dropped and reached progressively lower values similar to those at day 0 at the end of lactation. During pregnancy Body Weight was in low correlation with leptin (r = 0.32, P < 0.01) and there was no correlation with insulin (r = 0.13, P > 0.1); moreover, no correlation was found during lactation. T3 concentrations showed a U-shaped trend with a significant reduction at day 75 (P < 0.05). T4 concentrations fluctuated without any significant change (ranging from 27.0 ± 1.6 nmol/l to 32.1 ± 1.9 nmol/l). Cortisol concentrations ranged within the reference values (minimal value at day 30 (49.8 ± 6.3 nm/L) and maximal value at day 45 (72.5 ± 16.1 nm/L)). Our study states that canine pregnancy and lactation evoke many hormonal changes necessary to mother and fetuses. In particular, this is the first report regarding the pattern of leptin during whole pregnancy and lactation. The present work lays the groundwork for further studies on endocrine homeostatic mechanisms using the dog as a model for human medicine.

Pregnancy-associated glycoproteins (PAGs) concentrations in water buffaloes (Bubalus bubalis) during gestation and the postpartum period.

For the first time in literature this study describes the pregnancy-associated glycoprotein (PAG) profile of buffalo cows during gestation and the post-partum period using antiserum raised against PAG-molecules purified from buffalo placenta (AS#860). Ninety-eight buffalo cows, belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program, were enrolled in this study. Blood samples were taken on days 0 (AI), 23, 25, 28, 30 and then biweekly until the end of pregnancy. Pregnancy was confirmed by ultrasonography on days 28 and 45, and by rectal palpation from day 60 onwards. Blood samples were suspended for the non-pregnant cows on day 45, while the blood of 20 buffaloes that had calved was tested every five days from the day of calving until day 50 post-calving. A cut-off value of 1.0 ng/mL was used in order to discriminate between pregnant and non-pregnant buffaloes. We used Linear Mixed models after Log(x+1) transformation to analyse the PAG concentrations. Fifty-two buffalo cows had become pregnant out of 98 synchronized (53%) and 46 remained non-pregnant (47%) as shown by ultrasonography and the PAG analysis. Significant differences (P < 0.001) in PAG concentrations were observed between the pregnant and non-pregnant buffaloes from day 23 as the PAG of the non-pregnant cows was always close to zero. Conversely, the PAG of the pregnant cows increased progressively from day AI until day 105 post-insemination and then stabilized until the end of pregnancy. Regarding pregnancy diagnosis, the sensitivity of PAG-RIA 860 system (ability of the test to correctly identify pregnant buffalo) ranged from 23% on day 23-98% on day 28 post AI; the specificity (ability to correctly identify non-pregnant buffaloes) was 100% throughout the sampling period. PAG progressively decreased from parturition to day 25 post-partum; from day 30 post-partum, the concentrations fell below 1 ng/mL and were close to 0 on the last day of observation (50 d post-partum). In conclusion, our results showed that RIA-860 is highly accurate for diagnosing pregnancy in buffaloes starting from day 28 of gestation. Furthermore, the rapid disappearance of PAG concentration after calving means that a cut-off limit in post-partum for detecting a new pregnancy is not required.

Genetic diversity and antimicrobial resistance profiles of Campylobacter coli and Campylobacter jejuni isolated from broiler chicken in farms and at time of slaughter in central Italy.

AIMS: Genetic diversity and antimicrobial resistance of Campylobacter coli and Campylobacter jejuni were investigated along the broiler chicken production chain in central Italy. METHODS AND RESULTS: Campylobacter sp. isolated from cloacal swabs in farms (n = 116) and from the neck skin of chilled and eviscerated carcasses at slaughter (n = 24) were identified as C. coli (n = 99) and C. jejuni (n = 41) by multiplex PCR. Characterization by single amplified fragment length polymorphism (s-AFLP) revealed a specific genotype of Campylobacter for each farm. Minimal inhibitory concentration showed high prevalence of fluoroquinolones (70%), tetracycline (70%) and erythromycin (30%) resistance among C. coli isolates. Campylobacter jejuni isolates showed lower prevalence of fluoroquinolone (39%) and tetracycline (10%) resistance, and all isolates were susceptible to erythromycin. The S-AFLP types of the C. coli and C. jejuni isolates were associated with their antimicrobial resistance profiles (P < 0·001). CONCLUSIONS: The genetic diversity detected in Campylobacter isolates suggested that a specific genotype was harboured in each farm. A considerable number of C. coli isolates were resistant to erythromycin. SIGNIFICANCE AND IMPACT OF THE STUDY: Campylobacter coli was detected more frequently than C. jejuni in contrast to common findings for poultry. The high prevalence of 30% resistance to erythromycin in C. coli strains isolated from poultry is worrisome, as this is the first antibiotic of choice to treat human campylobacteriosis.

Using real-time PCR to identify pregnancy-associated glycoprotein 2 (PAG-2) in water buffalo (Bubalus bubalis) blood in early pregnancy.

This study investigates for the first time mRNA pregnancy-associated glycoprotein 2 (PAG-2) expression in blood cells during early pregnancy in water buffalo. The PAGs constitute a large family of glycoproteins expressed in the outer epithelial layer of the placenta in eutherian species. All PAGs are not concomitantly expressed throughout pregnancy; some of them are expressed in the earlier phases, whereas others appear later and are expressed over a shorter period. Twenty-one lactating buffaloes were analyzed-17 females were synchronized with PRID and artificially inseminated (AI), whereas four females were synchronized but not inseminated (control group). Blood was collected at Days 0, 18, 28, 40, and 75 from AI (AI = Day 0). Expression of PAG-2 mRNA in blood samples was measured with real-time polymerase chain reaction. Pregnancy diagnosis was performed on Day 28 (D28) and Day 40 (D40) after AI by ultrasonography (US) and by PAG-1 RIA method. The females diagnosed pregnant at D28 and confirmed pregnant at D40 were defined as D28(+)D40(+) group; the females diagnosed pregnant at D28 but not confirmed pregnant at D40 were defined as D28(+)D40(-) group; and the females that were diagnosed as nonpregnant on either days were defined as D28(-)D40(-) group. PAG-2 mRNA at Day 0 was not observed in any groups. The D28(+)D40(+) group showed the highest expression, starting on Day 18 and increasing progressively up to Day 75. PAG-2 mRNA was also expressed on Day 18 in both D28(+)D40(-) and D28(-)D40(-) groups, but their levels were lower than those of D28(+)D40(+) group and almost constant over time. PAG-2 mRNA was never detected in the control group. The significant difference in the expression of PAG-2 mRNA between the D28(+)D40(+) group and the D28(-)D40(-) group, starting from Day 18, suggests that these animals might have conceived, but have experienced early embryonic loss; therefore, the PAG-2 mRNA was still present in blood circulation although at lower levels, as found in the D28(+)D40(-) group. In conclusion, this study shows that PAG-2 mRNA can be detected in peripheral maternal blood cells earlier than circulating PAG-1 molecules and could be useful for studies on early pregnancy and embryonic mortality.

Addition of different concentrations of prostasome-like vesicles at neutral or slightly alkaline pH decreases canine sperm motility.

Prostasome-like vesicles (PV) are components of the canine prostatic fluid during ejaculation and despite some enzymatic activities have been identified recently, their functions in the reproductive events in this species are still poorly understood. In this study, we evaluated at neutral or slightly alkaline pH, the effects of different concentrations of purified PV on sperm cell motility characteristics and the impact on the short- and long-term preservation of preserved semen maintained at room temperature. Two different experiments were performed. In the first experiment, purified PV were added at increasing concentration (1.25, 2.5, 5, 10, and 20 µL equivalent to 1, 2, 4, 8, and 16 µg of protein, respectively) to aliquots of 100 µL of preserved semen maintained at 22 °C at the following time points: 0, 30, 60, 120, 240, and 480 min. Computer-assisted sperm motility characteristics and pH were assessed three times at each time points, for each sample and for every concentrations. In the second experiment, the purified PV were added with the same methods as described above but only at time 0. Sperm motility characteristics and pH were assessed over the time. This study showed how the addition of purified PV to preserved semen affects negatively (p < 0.05) at neutral and alkaline pH, both total and progressive motility in a concentration depending manner. Furthermore, prostasome addition was demonstrated to change the quality of sperm movement which may represent a mechanism facilitating sperm cells attachment to the uterine epithelium and facilitating energy preservation before fertilization.

Feline Injection-Site Sarcoma.

Feline injection-site sarcoma (FISS) is an aggressive tumor believed to arise from the proliferation of fibroblasts and myofibroblasts in areas of chronic inflammation, particularly at sites of injection. Local recurrence is frequent after surgical excision. Gelatinases (MMP-2 and MMP-9) and their inhibitor (TIMP-2) are endopeptidases pivotal in extracellular matrix remodeling and therefore in tumor invasiveness. The aim of this study was to investigate the immunohistochemical expression of MMP-2, MMP-9, and TIMP-2 in FISS to assess their usefulness as prognostic factors. Size, soft tissue sarcoma (STS) grading system, depth of infiltration, surgical margins, and Ki-67 index were evaluated as additional prognostic markers. Twenty-four cases of primary FISS were classified according to clinical follow-up as nonrecurrent (NR, n = 14; 58.3%) and recurrent (R, n = 10; 41.7%). MMP-2, MMP-9, and TIMP-2 were variably expressed in the FISS examined, confirming their role in tumor invasiveness, yet they did not show significant differences between the R and NR groups. These results could be due to different tumor stages or to the multiple activities of these enzymes, not limited to ECM remodeling. The immunohistochemical expression of these enzymes considered alone does not seem to be useful as a prognostic marker. STS grading system, depth of infiltration, surgical margins, and Ki-67 index did not relate to recurrence. Instead, the size of the tumor, measured after formalin fixation, with an optimal cutoff of 3.75 cm (accuracy = 86%; P < .05), and the mitotic count, with an optimal cutoff of 20 mitoses/10 HPF (accuracy = 80%; P < .05), could be evaluated as useful prognostic markers.

Retrograde maxillary nerve perineural injection: A tomographic and anatomical evaluation of the infraorbital canal and evaluation of needle type and size in equine cadavers.

The aim of this study was to investigate a new approach for equine maxillary nerve blocks, which can facilitate several orofacial surgeries. Current techniques aim at the maxillary foramen and approach via the zygomatic arch, conferring the risk of injury to several delicate structures in the target area. To investigate the feasibility of a retrograde approach from the infraorbital foramen, an anatomic study of the infraorbital canal and its surrounding structures was performed on 13 cadaveric skulls using computed tomography and anatomical dissection. Measurements included canal length and volume, its conformation and relationship with the enclosed structures, and infraorbital foramen diameters. The technical approach to simulate the distribution of local anaesthetic within the infraorbital canal was further defined, including needle selection among seven different needles, evaluating ease of insertion, trauma to surrounding tissues and spread of contrast medium toward the target area. To validate the technique, two Tuohy needles were randomly inserted at 12 infraorbital foramina and 10 mL of contrast medium was injected. CT verified the spread of the solution and possible complications. Each canal had a serpentine-curved pathway. Anatomical dissections evidenced gaps between the infraorbital nerve, vessels and the infraorbital canal. The integrity of these structures had been preserved from the passage of the selected Tuohy needles. This study suggests a feasible approach to the maxillary nerve block within the infraorbital canal providing an appropriate needle selection and technique to limit complications.

Probiotic mixture supplementation in the preventive management of trinitrobenzenesulfonic acid-induced inflammation in a murine model.

Inflammatory bowel diseases (IBD) are characterized by inflammatory conditions of the intestine. Probiotic bacteria (PB) can have beneficial effects in several gastrointestinal disorders. The objectives of this study were: (i) to provide an acute experimental IBD model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) in CD-1 mice, and (ii) to assess the preventive effects of Citogenex (Lactobacillus casei and Bifidobacterum lactis) supplementation on intestinal tissues and microbiota. Mice were inoculated intrarectally with saline, ethanol or different TNBS solutions. 1%TNBS induced clinical signs of colitis (P less than 0.01) and histological damage (P less than 0.01). Based on these results, mice were pre-treated with Citogenex or saline for 1, 2 or 3 weeks before 1%TNBS treatment. Probiotic pre-treatment determined a reduction of clinical signs (P less than 0.05), histological alterations of colitis (P less than 0.05) and increased beneficial bacteria (P less than 0.05). This study confirms that TNBS-induced colitis in CD-1 mice is useful for studying the mechanisms involved in IBD pathogenesis, and pre-treatment with Citogenex prevents the intestinal damage induced by TNBS.

E-cadherin, N-cadherin Expression and Histologic Characterization of Canine Choroid Plexus Tumors.

Choroid plexus tumors (CPTs) are reported with an increasing incidence in dogs, and they call for a reexamination of histologic features and criteria of classification corresponding to their biological behavior. In this study, the human World Health Organization classification was applied to 16 canine CPTs, and the expression of molecules involved in neoplastic cell adhesion (E-cadherin, N-cadherin), invasion (doublecortin), and proliferation (Ki-67) was investigated. Mitotic index was found to be the main criterion for grading CPTs. Cell density and multilayering of papillae were also statistically associated with histologic grade. Intraventricular spread and parenchymal invasion was observed for tumors showing histologic benign features. E-cadherin was expressed in all CPT grades, independent of tumor invasion. N-cadherin immunolabeling was more expressed in grade I than high-grade CPTs, whereas doublecortin expression was not detected in CPTs. An increasing proliferative activity was observed in relation with histologic grade.

A retrospective study of canine prostatic diseases from 2002 to 2009 at the Alfort Veterinary College in France.

A retrospective study was used to investigate the incidence of prostatic diseases in a large population of dogs at Alfort Veterinary Hospital and to clarify epidemiologic features, which might be of a great help to veterinarians in managing and discriminating prostatic disorders. During the investigation period, a total of 72,300 male dogs (coming mainly from the Ile-de-France region) were registered in the Alfort Veterinary College database, and 481 of them (0.7%) were found to have prostatic disorder. The diagnosis was carried out on the basis of clinical signs and ultrasound findings. Among dogs experiencing a prostatic disorder, most frequently recorded diseases were benign prostatic hyperplasia (45.9%) and prostatitis (38.5%), followed by abscesses (7.7%), cysts (5.0%), neoplasia (2.6%), and squamous metaplasia (0.2%). Our study revealed an incidence of 0.3% of prostatic disorders observed in intact male dogs, except in the case of prostatic neoplasia. The mean age of the dogs experiencing prostatic disorders was 8.6 ± 3.2 years. This was significantly different (P < 0.001). Large dogs were significantly more affected by prostatic disorders (P < 0.05), except for prostatic neoplasia. A breed predisposition was suspected in German Shepherd (odds ratio [OR] = 2.1; 95% confidence interval [CI]: 1.5-2.9), Rottweiler (OR = 1.8; 95% CI: 1.2-2.7), American Staffordshire Terrier (OR = 3.8; 95% CI: 2.5-5.8), Berger de Beauce (OR = 3.7; 95% CI: 2.2-6.1), and Bernese Mountain Dog (OR = 2.5; 95% CI: 1.3-4.7).

Food restriction during pregnancy in rabbits: effects on hormones and metabolites involved in energy homeostasis and metabolic programming.

This study examined the effects of food restriction during rabbit pregnancy on hormones and metabolites involved in energy homeostasis and metabolic programming. Pregnant does were assigned to four groups: the control group was fed a standard ration while the others received a restricted amount of food (30% restriction) during early (0-9 days), mid (9-18 days), and late (19-28 days) pregnancy. The pregnancy induced a coordinated range of adaptations to fulfil energy requirements of both mother and foetus, such as hyperleptinaemia and hyperinsulinaemia, reduced insulin sensitivity, increased cortisol and non-esterified fatty acid. Food restriction altered leptin, insulin, T3, non-esterified fatty acids and glucose concentrations depending on the gestational phase in which it was applied. Collectively, present data confirm that the endocrinology of pregnancy and the adaptive responses to energy deficit make the rabbit an ideal model for studying nutritional-related disorders and foetal programming of metabolic disease.

Effects of a bacterial lipopolysaccharide on the reproductive functions of rabbit does.

Systemic and local infections and inflammations are known to cause infertility in humans and animals. However, the mechanisms by which infection/inflammation induces infertility are only partially known. The objectives of this study were: (i) to provide models of systemic (acute) and local (sub-acute) inflammation by intra-peritoneal injection or intra-cervical deposition of lipopolysaccharide (LPS) in the rabbit and (ii) to assess their effects on uterine tissues and sperm transport in the genital tract of rabbit does. Intra-peritoneal administration of different doses of LPS induced systemic effects such as fever, anorexia and changes in white blood cells (WBC) count. In our study, LPS inoculation (100µg/kg) produced an inflammation-like status that lasted for about 3 days, with minimal distress for the animals. Intra-peritoneal administration of LPS 60h before artificial insemination induced a rapid increase of IL-1ß concentrations. The intra-cervical inoculation of LPS did not show any systemic effects, as confirmed by the lack of changes in body temperature, feed intake and WBC count. Histological examination of uterine tissues showed an endometritis-like inflammation status in LPS-treated does, more severe in those inoculated intra-cervically. The number of spermatozoa recovered from uterine horns and oviducts of intra-cervically treated does was less than that retrieved from intra-peritoneally treated animals and controls. These results suggest (i) that sub-acute or acute inflammation may cause infertility by compromising the uterine environment and/or impairing sperm transport and (ii) that the LPS-induced -infection/inflammation experimental model is useful for studying the mechanisms involved in reproductive dysfunctions in the rabbit.