In a bid to improve the productive potentials of local turkeys in developing countries, a total of 240 unsexed day-old poults arranged in a 2 × 2 factorial layout into 4 treatments with two rearing systems (indoor and outdoor) and two plumage colours (white and black) were used for this study. Poults were brooded for 4 weeks followed by an acclimatization period of 2 weeks in the two different rearing systems before the commencement of the study which lasted 10 weeks. Each treatment consisting of 60 birds was further sub-divided into six replicates of 10 birds per replicate. Data obtained were subjected to Analysis of Variance in a Completely Randomized Design. Results on performance in the grower phase showed turkeys reared in indoor system recorded significantly (p<0.05) higher weight gain (29.39 vs. 105.19 g/bird/day) and daily feed intake (27.18 vs. 98.11 g/bird/day) compare to turkeys under outdoor system. In addition, weight gain was significantly (p<0.05) higher (29.16 g/bird/day) in turkeys with black plumage than (27.42 g/bird/day) recorded in turkeys with white plumage. However, in the finisher phase turkeys under outdoor system recorded significantly (p<0.05) higher weight gain than turkeys under indoor system. In the finisher phase, interaction effects showed best weight gain and Feed Conversion Ratio (FCR) (39.22 g/bird/day and 4.60) in white-plumaged turkeys reared in outdoor system. Turkeys under outdoor system also had significantly (p<0.05) higher back and spleen percentages. However, proportions of thigh were significantly (p<0.05) higher in turkeys reared indoor. In addition, white-plumaged turkeys recorded significantly (p<0.05) higher (21.07%) cooking loss than 14.58% recorded in turkeys with black plumage. In conclusion, improved weight gain with best FCR at finisher phase as well as highest spleen portion and cooking loss in thigh meat was obtained in white-plumaged turkeys reared in outdoor system.
Staphylococcus pseudintermedius (SP) has been associated with high antibiotic-resistance rates (e.g. methicillin) in European countries [1,3]. This condition could be also related to the ability to produce biofilm [2]. The aim of this study was to investigate the presence of methicillin-resistant SP strains and determine their ability to produce biofilm and some crucial virulence factors. Forty-two SP strains, previously determined as multi drug resistant (MDR) by the disk diffusion method using a panel of 17 antimicrobial agents, were selected from our collection and tested phenotypically for the minimum inhibitory concentration (MIC) of methicillin and genotypically for the presence of mecA and blaZ genes. The ability to produce biofilm was assessed phenotypically by two different assays: the Congo Red Agar plates (CRA) and the Microtitre Plate test (MtP) and genetically by the amplification of icaA and icaD genes. Three virulence factors genes coding for bicomponent leukocidin and enterotoxins (luk-I, seC, se-int) were searched. Twenty-three strains revealed a value of MIC for the methicillin greater than 128 µg/mL. The 83% were mecA-positive and 86% resulted blaZ-positive; all the strains positive for mecA were also positive for blaZ. All SP strains resulted biofilm-producers by MtP assay and classified as weakly producers (4.7%), moderate producers (47.6 %) and strongly producers (47.6 %). In contrast, only 35.7% of all strains were considered biofilm-producers by CRA method. The amplification of icaA and icaD gene occurred respectively in 66.6% and 97.6%; only one strain was negative for both genes. Almost all strains were positive for luk-I (95%), seC (74%) and se-int (84%). Our data reveal the pathogenicity potential of SP strains from dogs, suggesting that they could be considered zoonotic potential agents and confirming other previous studies [3-5]. Moreover could be observed a clear linkage between antibiotic-resistance and ability to produce biofilm.
The in vitro development of vitrified oocytes (VOs) is still suboptimal (Mandawala et al., 2016) and the traditional two-dimensional (2D) culture systems might not be adequate to fully exploit VOs potential. The use of three-dimensional (3D) follicle-like structures, i.e. a combination of granulosa cells (GCs) and semipermeable 3D matrices, could mimic the physiological microenvironment and enhance VOs maturation and embryo development. The aim of this study was to assess the steroidogenic ability (estradiol and progesterone secretion) of GCs encapsulated in 3D barium alginate microcapsules (follicle-like structure) compared to GCs cultured in a 2D monolayer and the maturation outcomes of VOs cultured in these systems. After purification (Simsek & Arikan, 2015), cat GCs retrieved from isolated ovaries were in vitro cultured for 6 days in 3D microcapsules (Vigo et al., 2005) or in 2D monolayers. On days 2 and 6, conditioned medium was collected and hormonal determination by enzyme-linked fluorescent assay was performed. On the same days, 3D and 2D cultured GCs were used as artificial milieu for in vitro maturation of VOs obtained by Cryotop protocol. Nuclear maturation was assessed by bis-benzimide staining. Steroidogenesis was observed in 3D follicle-like structures as well as in 2D monolayers; hormonal concentration increased over time and on day 6 it significantly differed between systems (p=0.02). Vitrified oocytes resumed meiosis in presence of GCs cultured for 2 days (3D: 45.5%; 2D: 56.7%), while GCs cultured for 6 days significantly hindered VOs meiosis progression in monolayers (21.7%, p=0.007), but supported high proportions of full maturation in follicle-like structures (26.7%, p=0.07). Granulosa cells in 3D microcapsules maintained their physiological features and these follicle-like structures were able to restore VOs developmental abilities. However, further advancements in VOs culture methods would optimize the use of these valuable resources.
Recently, many studies in livestock have focused on the identification of Copy Number Variants (CNVs) using high-density Single Nucleotide Polymorphism (SNP) arrays, but few have focused on studying chicken ecotypes coming from many locations. CNVs are polymorphisms, which may influence phenotype and are an important source of genetic variation in populations. The aim of this study was to explore the genetic difference and structure, using a high density SNP chip in 936 individuals from seven different countries (Brazil, Italy, Egypt, Mexico, Rwanda, Sri Lanka and Uganda). The DNA was genotyped with the Affymetrix Axiom®600k Chicken Genotyping Array and processed with stringent quality controls to obtain 559,201 SNPs in 915 individuals. The Log R Ratio (LRR) and the B Allele Frequency of SNPs were used to perform the CNV calling with PennCNV software based on a Hidden Markov Model analysis and the LRR was used to perform CNV detection with SVS Golden Helix software.After filtering, a total of 19,027 CNVs were detected with the SVS software, while 9,065 CNVs were identified with the Penn CNV software. The CNVs were summarized in 7,001 Copy Number Variant Regions (CNVRs) and 4,414 CNVRs, using the software BedTool.The consensus analysis across the CNVRs allowed the identification of 2,820 consensus CNVR, of which 1,721 were gain, 637 loss and 462 complex, for a total length of 53 Mb corresponding to the 5 % of the GalGal5 chicken autosomes. Only the consensus CNV regions obtained from both detections were considered for further analysis.The intersection analysis performed between the chicken gene database (Gallus_gallus-5.0) and the 1,927 consensus CNVRs allowed the identification (within or partial overlap) of a total of 2,354 unique genes with an official gene ID. The CNVRs identified here represent the first comprehensive mapping in several worldwide populations, using a high-density SNP chip.
Copy Number Variations (CNVs) have become promising markers, representing a major source of genomic variation. CNV involvement in phenotypic expression and in different diseases onset have been widely demonstrated in humans as well as in many domestic animals. However, this genomic investigation is still missing in Felis catus. This work is the first CNV mapping from a large data set of Whole Genome Sequencing (WGS) data in the domestic cat. A total of 42 cats of 14 different breeds were sequenced on the Illumina XTen (Washington University-St. Louis) which generated approximately 30-fold genome coverage from 150 paired-end reads (99 Lives Initiative). Maverix Biomics mapped the reads on the v6.2 reference assembly. CNV detection was performed using cn.mops and CNVnator, two Read Depth method software. One cat was excluded as outlier while, on the 41 remaining individuals, 1640 CNVs were detected by both the software and used to obtain 2891 CNVRs with BedTools. CNVRs covered the 0.4% of the total cat genome, with 136 loss, 127 gain and 26 complex detected (Fig. 1). A total of 164 singletons were identified and 9 CNVRs mapped in at least the 50% of the individuals. The number of CNVs in each cat ranged from 12 to 83. The clustering analysis of the detected CNVs was performed with R package “pvclust” and shows that same breed individuals cluster together. This study has led to the genetic characterization of 14 main cat breeds. Further analyses including other breeds and considering the genes located within these regions, could led to better evaluate the relationship between the presence of a specific CNV and a specific breed trait. This study can be considered a starting point for genomic CNV identification in the domestic cat, which could be further developed using the new released Felis catus vs9.0 reference aassembly.
Animal production systems produce large quantities of manure, which is recognized as a significant source of heavy metals (HMs) (Hejna et al., 2018). Some HMs are essential nutrients and zinc oxide (ZnO) was often used at high doses to control the enteric disorders mainly in the swine post weaning phase (Rossi et al., 2013, 2014). The general increase of HMs content was registered in the livestock output with a negative impact on the environment. Preliminary data showed that swine manure was an important source of Zn, Cu, Mn and Se to the environment reflecting the HMs content in feeds (Hejna et al., 2017a). The aim of this study was to evaluate the ability of Typha latifolia (TL) and Thelypteris palustris (TP) to bioaccumulate trace elements, from water as a cost-effective plant-based approach of wastewater remediation in pig livestock. The experimental design included four mesocosms (width: 4.0 m, length: 2.0 m, depth: 0.7 m; 695L of water, 210kg of soil): two controls, planted with TP (TPC) and TL (TLC) respectively and two treated, planted with TP (TPT) and TL (TLT) respectively. The treatment was represented by a mineral feed additive premix dissolved in TPT and TLT with the following final HMs concentration: Zn: 44.02mg/L; Cu: 8.63mg/L; Mn: 10.83mg/L; Se: 0.09mg/L. Such high concentrations, corresponding to polluted wastewater, would be sufficient to reach the potential saturation limit of the substrates in the short experimental period. At day 0 (T0), day 15 (T1) and day 45 (T2) samples of roots, leaves, stems, soil and water were collected, dried and principal chemical component were estimated according to the official method of Analysis of Association of Analytical Communities (AOAC). Samples were also mineralized by an ultravawe single reaction chamber and analyzed using inductively coupled plasma mass spectrometry (ICP-MS). Obtained results showed that TL and TP tolerated high levels of Zn, Cu, Mn and Se with no visual toxicity signs or significant effects on growth during the entire experimental period. TP appeared more effective than TL at translocating elements from water to plant tissues. In particular, TPT showed a significant increase of Zn and Cu content in whole plants from T0 to T2 (p≤0.001). At T2 the mean zinc concentration was 409.26±342.33 mg/kg d.m. and 271.64±64.85 mg/kg d.m. in TPT and TLT plants respectively. In the plants of control mesocosms, differences between T0 and T2 were not observed. Results suggested that both plant species were able to reduce the available amount of metals from the contaminated wastewater, thus TL and TP plants may be candidates for the phytoremediation approach to control HMs output from the livestock wastewater. Acknowledgements: supported by MIPAAF 2015.
Predictable immobilization of wild zebras is challenging and there is massive variation in opiate response within different species. Etorphine combined with azaperone is considered the protocol of choice, but no studies have investigated the physiological response to this procedure of immobilization in plains zebras. Eleven free-ranging plains zebras (Equus quagga) were immobilized in Kenya using a combination of etorphine 0.019 ± 0.003 mg/kg and azaperone 0.27 ± 0.05 mg/kg administered intramuscularly with a projectile dart. After recumbency, an arterial sample was performed for blood gas analysis and physiological parameters were recorded every five minutes. Descriptive scores were given to the exertion resulting from high-speed chasing and to the quality of induction, immobilization and recovery. Diprenorphine or naltrexone were used for opioid antagonism. In all zebras, the combination induced quick inductions within 3.5 ± 0.8 minutes and provided reliable recumbencies without attempts to stand for the entire duration of the immobilization. The average heart rates, respiratory rates and mean arterial blood pressure recorded were 102 ± 42 beats/minute, 18 ± 4 breaths/minute and 145 ± 28 mmHg respectively. Arterial gas analyses demonstrated mild to severe and partially compensated metabolic acidosis and hypoxia, while electrolytes were within equids range. In particular, higher exertion levels during the chasing were significantly correlated to worse immobilization scores (p=0.008) and hyperthermia occurrence (p=0.0012) and non-significantly to more severe acidosis. Recoveries from anaesthesia were smooth, on average 121 ± 38 seconds after diprenorphine/naltrexone administration. Etorphine-azaperone combination produced physiological alterations in free-ranging plains zebra such as tachycardia, hypertension, metabolic acidosis and hypoxemia. However, these preliminary results indicate that high-speed chase might be responsible for the physiological imbalance and that this drug combination does not suppress the compensatory response. Regardless of the metabolic status, recover from immobilization was uneventful and all zebras went back to normal behavior thereafter.
The study was carried out to investigate the effect of feeding broiler chicken on different vegetable oils with feed additives on the quality characteristics of chicken meat. A total of 216 one-day-old chicks of ( Hubbard ) strain were randomly assigned to six dietary treatments as (2×3) factorial designs where two sources of dietary oil with three levels of commercial multi-enzyme feed additives. Treatments were: soybean oil only (T1), soybean oil+ ZAD (T2), soybean oil+ AmPhi-BACT (T3), palm oil only (T4), palm oil + ZAD (T5) and palm oil + AmPhi- BACT (T6). Results showed that feeding broiler chicken on different types of dietary oils had significant effect on the fatty acid profile of broiler chicken meat. UFA/SFA ration of broiler chicken groups (T4, T5adT6) were significantly lower compared with (T1, T2 and T3) groups. Broiler fed on soybean oil had significantly higher n-6: n-3 ration compared with broiler fed on palm oil. Regardless of the source of dietary oil, significant differences were observed in the most of fatty acid profile in the chicken meat among levels of commercial multi- enzyme feed additives. Meat of T5and T6 had the higher pH value, followed by meat of T1and T3 groups, while the lowest pH value found in meat of T2 and T4. The higher cooking loss was found in meat of T4 while, meat of T5had the lowest value. Data of chilling loss indicated that the differences between dietary treatments were not significantly different except for meat of T6 which had the higher chilling loss. No significant differences were found in color measurements between dietary treatments.
In livestock, genomics has been used since a decade in combination with phenotypic information for the estimation of breeding values. In honey bees (Apis mellifera), the advantage for including genomics in selective breeding programmes is represented by the possibility to reduce the generation interval and increase the accuracies of estimated breeding values resulting in higher genetic gain (Brascamp et al., 2018). The limit for this application is DNA extraction. Extraction methods for small animals such as insects often rely upon destructive approaches. The challenge is to develop tissue sampling methods that permit the survival of the animal while providing adequate quality DNA for genotyping. Along with previous reports of DNA extraction from several matrices, this study aims to contribute in developing suitable methodologies for genotyping honey bees queens using DNA extracted from wing cuttings (Chaline et al., 2004; Gregory and Rinderer, 2004; Gould et al., 2011). The clipping of the queen wings in beekeeping is a common practice and it ensures the survival and normal activities of the animal (Forster, 1971). A total of 57 queens with known pedigree were enrolled for this study. Wings from each queen were cut and stored at -20°C until processed (Fig. 1). Extractions were carried out using a modified protocol provided by Qiagen (DNeasy® Blood & Tissue). The modification consists in an initial incubation of the samples with proteinase K for 20 minutes, further steps are carried out following the manufacturer’s instructions. To test the suitability of the extracted DNA for genotyping, PCR was performed on Esterase FE4 like gene. Although quantification with NanoDrop™ resulted in <20 ng/μL of DNA in solution, the extracted material was sufficient for PCR amplification of candidate genes for sequencing and genotyping. Our results show that it is possible to extract DNA from wings’ cuttings permitting to implement genomic approaches in honey bee selective breeding. Further work will analyse possible association between genetic variability and phenotypes of interest.
The aim of this study was to evaluate the effect of Astragalus polysaccharide (APS) supplementation on the growth performance, plasma biochemical parameters, and plasma immune and antioxidant indexes of weaned piglets in a commercial swine farm. A total of 120 piglets weaned at 22 days and allocated to 2 groups, and fed a basal diet either without (CTR) or with 200 mg/kg of APS in a local commercial farm for a 42-d experiment. At end of the trial, one piglet from each pen was selected for blood sampling. The results showed that dietary APS decreased the feed conversion ratio (FCR) compared to the CTR group from day 14 to day 28 and day 0 to day 42 (P = 0.08 and 0.02, respectively). In addition, supplementation of APS had the tendency to increase the plasma superoxide dismutase activity and IgG content of piglets compared to the CTR group on day 42 (P = 0.06 and 0.09, respectively). Results in this study suggested that dietary APS might have a beneficial effect on growth performance and health status of weaned piglets under the commercial condition.
Mycobacteria in cats are responsible for cutaneous or visceral disease, often with systemic involvement (Gunn-Moore et al., 2011). Infection can be sustained by tuberculous (e.g. M.bovis) and non-tuberculous mycobacteria (e.g. M.avium and M.lepraemurium) (Lee et al., 2017). The aim of this study is to evaluate the distribution of the lesions in feline visceral mycobacteriosis. Twenty-nine necropsy cases of feline visceral mycobacteriosis, conferred from 1965 to 2017, were studied. On histopathology hematoxylin-eosin and acid-fast stains were performed. Mycobacteria strains were identified by microbiological and molecular methods. Twenty-three out of 29 cases were submitted for necropsy in autumn and winter. When breed was known: nine cats were Persian, 8 Siamese and 10 crossbreed. In 17 out of 24 cases where age was known, it was lesser than 5 years. Two cases had lesions confined to the digestive system, 3 to the respiratory system, while 24 cases were systemic forms. Thirteen cases out of 29 were identified: 2 as Mycobacterium spp, 7 as M.bovis and 4 as M.avium. Siamese, Persian breeds and young cats were overrepresented as already described (Gunn-Moore, 2014). The occurrence of mycobacterioses in our cases was higher during cold seasons, contrary to the man in which tuberculosis is a long term localized disease, more often diagnosed in summer even if acquired in winter; in cat the disease tends to generalize making shorter the course of the disease (Fares, 2011). This is confirmed by lesions’ distribution that indicates a greater occurrence of systemic compared to localized forms. Features of gastrointestinal lesions indicate the alimentary as the primary route of infection (Fig.1). Since the most recent cases were sustained exclusively by M.avium, while older cases were caused by M.bovis, a switching in the most diffused species or a different source of infection might occurred in the last years (Pesciaroli et al., 2014).
MicroRNAs (miRNAs) are a class of short non-coding RNA, which interact with the 3’ UTR region of complementary mRNA to decrease or inhibit the translation of proteins (Lai, 2002). MiRNAs regulate pathways in various pathophysiological status, and are regarded as biomarkers for early diagnosis of several diseases, including cancer (Di Leva et al., 2014). The study aims to evaluate the quality and purity of miRNAs extracted from a) 11 archival Formalin Fixed and Paraffin Embedded (FFPE) samples of Mast Cell Tumour (MCT) at stage I, II, III and IV, and 8 intra-patient healthy controls; b) samples collected during surgery, including 6 samples of saliva, primary tumour biopsy and serum/plasma. The quality of miRNA largely influence the downstream experiments, and must be carefully evaluated before performing for examples, the sequencing reaction. MiRNA extraction was carried out using commercial kits (Qiagen) and quantify using Small RNA Kit (Agilent) on Agilent 2100 Bioanalyzer. The results showed that the concentration of miRNAs from FFPE, saliva, primary tumor biopsy and serum was acceptable with a Median (Me)= 56,91 ng/ml, Me=10,30 ng/ml, Me=3,44 ng/ml and Me=0,71 ng/ml, and a miRNA/Small RNA ratio of 48%, 61%, 17% and 76%, respectively. The concentration of miRNAs from plasma was not detectable. Studies reveal that plasma ranks as the first choice source for diagnostic purpose, much more than serum (Aung et al., 2014), but the debate remains open and subsequent analyses are needed. The concentration of miRNAs from FFPE and saliva samples is higher than that from other matrices. Possible explanations include a) different quantity and quality of starting materials; b) nucleic acids fragmentation, due to the formalin fixation and paraffin embedded procedure; c) presence of nucleases in saliva, which produce small fragments recognized as miRNAs or smallRNAs. In conclusion, the quantity and the purity of miRNAs, obtained using Qiagen commercial kits, are reliable for further NGS analysis.
Introduction: Ticks are haematophagous ectoparasites of vertebrates habitually parasitizing avian species, which may contribute to tick dispersal across continents during migrations (Hasle 2013; Altizer et al., 2011). Midichloria bacteria can be transmitted to the vertebrate host during the tick bite (Bazzocchi et al., 2013; Serra et al., 2018). Although many avian species are common hosts of ticks harbouring Midichloria (e.g. Ixodes, Hyalomma ), the circulation of this bacterium in birds has never been investigated. The aims of this study are: 1) evaluate the presence of Midichloria DNA in H. marginatum ticks and blood collected from trans-Saharan migratory birds; 2) quantify Midichloria bacteria in ticks through a novel quantitative PCR (qPCR). Material and methods: A total of 256 H. marginatum ticks and 97 blood samples were collected from three different migratory species ( Phoenicurus phoenicurus, Saxicola rubetra and Sylvia communis ) on Ventotene Island (Central Italy) and DNAs were extracted. A nested-PCR targeting the 16S rRNA gene of Midichloria was used to detect bacterial presence. Subsequently, primers targeting the gyrB gene of Midichloria and the cal gene of H. marginatum were designed and used in a qPCR for Midichloria quantification. Results were expressed as gyrB / cal copy numbers ratio. Results and discussion: 94% of Hyalomma ticks harbored DNA of Midichloria belonging to the monophylum associated with ticks, while the bacterial DNA was detected in 44.3% of blood samples. Furthermore, engorged ticks showed significantly higher bacteria load than unengorged ticks (Table 1; Wilcoxon sum-rank test: z=3.14; p=0.0017), similarly to what has been observed for M. mitochondrii in I. ricinus ticks. Conclusions: This work provides evidence for the presence of circulating Midichloria DNA in long-distance migratory birds, suggesting an enhanced worldwide spread of these bacteria across haematophagous ectoparasite populations. Future studies are necessary to increase the knowledge of Midichloria role in the biology of this tick species.
Following the concepts of precision feeding, the right components balance (Sova et al., 2014) and the correct particle size distribution (PSD, Khan et al., 2014) of total mixed ration (TMR) are essential for a complete homogeneity of the diet and are strongly influenced by adopted mixing time (MT, Humer et al., 2018, Schingoethe et al.,2017). The aim of the trial was to determine the influence of two MTs (MT1≤7min and MT2>7min) on the chemical homogeneity and PSD along the feeding alley. Diets were performed with a horizontal cutter-mixer wagon (Gulliver 4016, Sgariboldi), and TMR samples were collected from the beginning, middle and end of the feeding alley after discharge. Triplicate samples of the diet were collected for chemical composition analyses (moisture, CP, Ash, EE, NDF and ADF) and PSD evaluation (Heinrichs and Kononoff, 2002) over two months (two sampling/week). Statistical analysis was performed by a PROC MIXED for repeated measurements of SAS. MT1 evidenced a non-uniform distribution of moisture content along the feeding alley (P=0.05): lower moisture was found at the end than at the beginning and in the middle (47.55 vs 51.13 and 51.00%, respectively; P<0.01). No significant effects of MTs were recorded for other chemical parameters. The PSD showed trend to a higher retained amount of fibre in MT1 upper sieve (14.79 vs. 10.14%; P=0.06), while lower amount of feed was found in middle and bottom sieve than MT2 (38.9 and 12.81 vs 42.17 and 14.32%, respectively; P=0.08 and P=0.06). With respect to TMR distribution along the feeding alley, no differences were found between MT1 and MT2. Day of sampling evidenced significant variation both in chemical and physical composition (P<0.05). Obtained preliminary data evidenced the influence of MTs on composition and on PSD of the provided diet; results suggest to daily measure moisture of raw material in order to avoid negative changes in dry matter intake.
Cytological evaluation of splenic lesions is a routine preoperative diagnostic technique. However, few studies have evaluated the utility of diagnostic cytology in canine splenic diseases (Ballegeer et al., 2007; Christensen et al., 2009; Watson et al., 2011). Our aim was to evaluate accuracy, sensitivity, specificity, positive and negative predictive value of cytology to diagnose canine splenic conditions using histopathology as the gold standard. Splenic cytological samples obtained between January 1998 and 2018 were retrospectively evaluated. Cases were included only when cytology and histology of the same lesion were available. All samples were blindly reviewed. Ninety-two cases were included (65 neoplasms, 27 non-neoplastic lesions) and classified as: 36 true positive, 29 false negative, 26 true negative and 1 false positive. Splenic cytology had a diagnostic accuracy of 67.39%, a sensitivity of 55.38%, a specificity of 96.3%, a positive and negative predictive value of 97.3% and 47.27% (Tab.1). To our knowledge, this is the first study reporting conjunctively accuracy, sensitivity, specificity, positive and negative predictive value of cytology in the diagnosis of canine splenic disorders. The major limit of splenic cytology was a reduced sensitivity related to a high number of false negative results that strongly correlate with lesion distribution, size and type and with the blood storage function of the spleen resulting in hematic samples (Bertazzolo et al., 2005; O’Brien et al., 2013). Limitations were balanced by high specificity and positive predictive value making splenic cytology a valuable preliminary diagnostic tool to assist further diagnostic and therapeutic approaches in splenic disease.
Sturgeons, as well as paddlefishes, belong to the Acipenseriformes group, which is one of the most primordial 57 orders of the Osteichthyes that comprehends 25 species spread throughout Europe, Asia and North America. The present study aims at investigating muscle growth and development as well as fatty acid profile in Siberian sturgeon free-embryos when subjected to three different rearing densities. Fatty acids, in particular polyunsaturated fatty acids of n-3 series, are generally known as key nutrients in fish larvae.This study was approved by the Ethic Committee of the University of Milan (OPBA_22_2017). Siberian sturgeon larvae were reared at 18°C, at three stocking densities until complete yolk-sac absorption: low (LD, 30 larvae/l), mid (MD, 80 larvae/l) and high (HD, 150 larvae/l). Sampling timepoints were: hatching, schooling and complete yolk-sac absorption stage (YSA). Sacrificed larvae were weighed and histological analyses were performed in order to assess muscle development as described elsewhere ; fatty acid profile was determined by GC-FID analysis as described by Vasconi et al. (2015). Statistical analysis was performed with SAS software (v. 9.3, Cary Inc., NC).At the end of the experiment, LD larvae presented a higher weight than larvae reared at the other two densities (P<0.05). Within the schooling stage (Figure 1), Total Muscle Area was lower for HD larvae (P<0.05); red and white muscle areas in schooling and YSA were higher than at hatching (P<0.05), regardless the density. Concerning fatty acids, no statistical differences were recorded between different rearing densities, while during the development regardless the rearing density, there was a common pattern: linoleic and alfa linolenic acids, significantly decreased their relative content, while others, as arachidonic acid and DHA, significantly increased. Siberian sturgeon larvae reared at LD or MD reveal an anatomically normal muscle development, while in the HD it is possible to observe a slowdown.What the aquaculture industry requires is a set of guidelines that allows the development of a sustainable industry, so that we tried to develop guidelines for stocking density in the very early stage of farming. As a conclusion, it would seem that mid density could be more suitable for this species in this stage of development.
Quadriceps contracture is a debilitating and uncommon condition, mostly affecting young dogs. Itcan be congenital or acquired ) and is reported to induce muscular hypotrophy/fibrosis, progressive degenerative joint disease, bone hypoplasia and limb hyperextension. The aim of this study was to elucidate anatomic, tomographic and biomechanical features of stifles affected by quadriceps contracture. Seven 2-month-old dead Dobermann Pinschers with unilateral quadriceps contracture were included. Before gross anatomic evaluation, all stifles underwent Computed Tomography before and after intra-articular administration of iodinated contrast medium. Images were acquired in double positioning (stifle extension and flexion) to identify articular cartilage, Ossification Centres’ (OCs) and menisci abnormalities, which were compared between affected and unaffected limbs. In all affected limbs the stifle was back-turned, the distal femur was extra-rotated and the patella was luxated proximo-medially. Severe lack of physiological stifle movements (rolling, gliding, spinning) was observed, so that affected joints could not be flexed. The articular cartilage of the femur was flattened and irregular in thickness, the femoral trochlea was hypoplasic and sloping, the menisci were misshaped. The OC of the distal femur and proximal tibia were misshaped; the tibial plateau was oriented caudodorsally-cranioventrally and significantly smaller (P<0,05). Quadriceps contracture influenced stifle development. The action of quadriceps insertion on the tibia prevented normal development of the plateau, causing wedging and abnormal orientation. Constant compression also induced external rotation of the distal femur (unable to develop distally) and patellar luxation, ending up in genu recurvatum. Static compression was likely responsible for femoral trochlea hypoplasia, articular cartilage and meniscal deformation, due to the lack of physiological stifle movements. Quadriceps contracture induces severe alterations of stifle development in affected puppies. Histology, histochemistry and immunohistochemistry may better define the nature of such bone, cartilage and meniscal alterations.
Genetic selection of dairy cattle has classically been based on fertility and productivity (kilograms of milk, protein, fat). In recent years a growing interest in characters related to health and efficiency of the animals has taken hold. The selection of animals with a high feed efficiency can bring benefits in terms of health, productivity and environmental impact. A first batch of 16 Holstein heifers (mean age 12.63 ± 2.90 months) was selected and housed in a tie-stall of the Centro Zootecnico Didattico Sperimentale (CZDS) of Lodi for the whole length of the trial (35 days). Blood samples were collected and sent to the lab to perform genotyping of the animals. Heifers were fed a composed ration of sorghum silage and total mixed ration, delivered daily on individual feeders to ensure each animal continuous access to the feed. Animals had free access to the water. Feed intake was obtained weighing the supplied ration and the residual the day after. Individual body weight (BW), body condition score (BCS), hearth girth (HG) and height at the wither (W) were measured weekly. Feed Conversion Ratio (FCR) and Residual Feed Intake (RFI) were calculated using Dry Matter Intake (DMI) data. Fecal samples were collected weekly to assess any relation between feed efficiency and acidic profile of the feces. Preliminary results show an average daily gain of 941 ± 206 grams, an increase of mean BW from 374.63 ± 17.06 (day 0) to 407.56 ± 17.06 kilograms (day 34), an increase of mean BCS from 3.97 ± 0.11 (day 0) to 4.19 ± 0.11 (day 34), an increase of mean HG from 169.25 ± 2.83 (day 0) to 177.41 ± 2.83 centimeters (day 34), an increase of mean W from 127.28 ± 1.23 (day 0) to 132.69 ± 1.23 centimeters (day 34).
Organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) are synthetic chlorinated compounds classified as POPs whereas only the penta e tetra-brominated polybromodiphenyl ethers (PBDEs) are so defined by the Stockolm Convention (Stockholm Convention, 2005) in order to elimitate or restrict the use of POPs. Organophosphorus insecticides (OCPs) represent important environmental and food contamination sources, widely used in agriculture. Among polyciclic aromatic hydrocarbons (PAHs), benzo[a]pirene is classified by IARC in Group 1, as cancerogen and Benzo[a]fluoranthene as a Group 2B, as possible cancerogen (IARC, 2012; IARC, 2010). EFSA (European Food Safety Authority) has released a scientific opinion on the risks to public health related to the presence of brominated flame retardants in food (EFSA, 2011) and in 2014 European commission has asked Member States to monitor the presence of brominated flame retardants (BFRs) in food over the next two years (EC, 2014). Due to their heir n-octanol/water partition coefficient (log Kow), they accumulate in fat tissue, bioconcentrate and biomagnify in the animals at the higher trophic levels, possibly causing, through chronic exposure, endocrine disruption and cancer (Wania et al., 1995; Vallack et al., 1998). The aim of this study is to evaluate the presence of OCPs, PCBs, PBDEs and PAHs in chamois and wild boar from Eastern Piedmont, Italy. A total of 20 chamois and 20 wild boar muscle samples were collected during the hunting season 2017, from Verbania Cusio Ossola (VCO) (Fig 1). The chemical analysis for the detection of OCPs, PCBs, PBDEs, and PAHs was performed by GC-MS/MS on muscle samples purified and extracted using a QuEChERS technique, validated according to SANTE 2017 (SANTE/11183/2017). These preliminary results show the ubiquitary presence of the studied contaminants. PCBs have been found more in chamois (45%) than in wild boar (35%). No PBDEs were detected in wild boar but in chamois were found with a prevalence of 35% and concentration 0.25-1.52 ng g-1. About OCPs, phorate and demeton were found in wild boar (55%-15%) and chamois (32%- 35%) with range concentrations 0.21-20.1 ng g-1. No PAHs were detected, expect antharacene for one samples in wild boar (0.53 ng g-1). Further studies are in progress in order to correlate environmental contamination and game animals.
Knowledge of host-pathogen interactions at molecular level is crucial for understanding of pathogenesis, disease prevention and cure. Here Borrelia is presented as a model neuroinvasive pathogen which employs vast immune evasion mechanisms like - drastic change in antigenic proteins, complement regulatory protein binding, antigenic variation etc. It is proposed that Borrelia may employ multiple strategies to evade host's complement system by binding complement regulatory proteins like factor H, vitronectin, C4BP and CD59. Amplitude of these mechanisms, their pathogen species dependent disparity, and characterization of interacting proteins from both sides (host and pathogen) will be presented during VAS presentations. Borrelia is also capable of invading central nervous system (CNS) in hide in the immune privileged site – the CNS. To do so it must cross the blood brain barrier (BBB). The mechanisms of the BBB crossing of this organism, like many of the other CNS invading pathogens, are still subject of on-going research. Our decade of research indicate the paracellular mechanism of the BBB penetration of Borrelia. Paracellular penetration of pathogen needs multiple protein:protein interactions between pathogen surface proteins and endothelial cells. The second part of my presentation in VAS will be dedicated to mechanisms employed by Borrelia to modulate cell signaling events in brain microvascular endothelial cells to cross the BBB. This presentation will also give sneak peak of several state-of-the-art technologies in genomics and proteomic used to understand the mechanisms of neuroinvasive and complement evasion by Borrelia. I hope that doctoral fellows will take advantage of the experimental pipeline that will be presented in VAS and apply successfully in their research
