C. Dieci, V. Lodde, Rémi Labreque, I. Tessaro, M. Sirard, A. M. Luciano
Oocyte’s chromatin gradually becomes more compacted during the final stage of oocyte development and the level of chromatin compaction is considered a marker of oocyte differentiation [Luciano et al, 2014]. Moreover, several studies demonstrate that in vitro pre-maturation treatments (Pre-IVM), aimed to improve the developmental capability of immature oocytes, might behave differently depending on the oocyte metabolic status, when it is isolated from follicle [Luciano et al., 2011]. This study aims at identifying correlations between cumulus-oocyte complex (COC) morphology and oocyte chromatin configuration and secondly at testing the hypothesis that only fully grown oocytes at earlier stages of differentiation with loosely compacted chromatin (GV1) can benefit from Pre-IVM treatment. COCs were collected from bovine 2-6mm ovarian follicles, and further divided in three groups according to their morphology (Class-1, 2 and 3) as previously described [Blondin & Sirard, 1995]. Analysis of chromatin configuration revealed that only Class-1 COC was enriched in GV1 oocyte, while Class-2 and 3 presented a similar distribution of GV1, GV2 and GV3 oocytes, where GV2 and 3 oocytes are characterized by increased chromatin compaction [Lodde et al., 2007]. Then COCs were divided into two groups, one containing Class-1 COCs and the other containing Class-2 and 3 COCs and subjected to pre-IVM for 6 hours in presence of cilostamide and 10-4 UI/ml rhFSH. Finally, COCs underwent standard in vitro maturation (IVM) for 22 hours, in vitro fertilization and embryo culture. Blastocyst rate and embryos cell number were assessed at day 7. Pre-IVM positively affected developmental competences of Class-1, while in Classes 2 and 3 Pre-IVM had detrimental effects.In conclusion COCs morphology could be used as a non-invasive approach to select population of oocyte with different cultural needs. These data could be useful in setting-up dedicated IVM protocols considering specific genes and pathways to improve IVP efficiency.
在卵母细胞发育的最后阶段，卵母细胞的染色质逐渐变得更加致密，染色质致密程度被认为是卵母细胞分化的标志[Luciano et al .， 2014]。此外，一些研究表明，体外预成熟处理(Pre-IVM)旨在提高未成熟卵母细胞的发育能力，但当卵母细胞从卵泡中分离出来时，其代谢状态可能会有所不同[Luciano et al.， 2011]。本研究的目的是确定卵丘-卵母细胞复合体(COC)形态与卵母细胞染色质构型之间的相关性，其次是为了验证一个假设，即只有处于分化早期、染色质松散致密(GV1)的完全成熟的卵母细胞才能从预ivm治疗中受益。从牛2-6mm卵巢卵泡中收集COCs，并根据其形态(1、2和3类)进一步分为三组，如前所述[Blondin & Sirard, 1995]。染色质结构分析显示，只有1类COC在GV1卵母细胞中富集，而2类和3类在GV1、GV2和GV3卵母细胞中分布相似，其中GV2和3卵母细胞的特征是染色质压实增加[Lodde等，2007]。然后将COCs分为两组，一组含有1类COCs，另一组含有2类和3类COCs，在西洛胺和10-4 UI/ml rhFSH存在下进行ivm前6小时。最后，COCs进行标准体外成熟(IVM) 22小时，体外受精和胚胎培养。第7天测定囊胚率和胚胎细胞数。预ivm对第一类学生的发展能力有积极影响，而对第二类和第三类学生的发展能力有不利影响。综上所述，COCs形态学可以作为一种无创的方法来选择不同培养需求的卵母细胞群体。这些数据可能有助于建立考虑特定基因和途径的专用IVM方案，以提高IVP效率。
{"title":"Morphological markers to select populations of oocytes with different cultural needs for dedicated pre-maturation protocols","authors":"C. Dieci, V. Lodde, Rémi Labreque, I. Tessaro, M. Sirard, A. M. Luciano","doi":"10.13130/2283-3927/8367","DOIUrl":"https://doi.org/10.13130/2283-3927/8367","url":null,"abstract":"Oocyte’s chromatin gradually becomes more compacted during the final stage of oocyte development and the level of chromatin compaction is considered a marker of oocyte differentiation [Luciano et al, 2014]. Moreover, several studies demonstrate that in vitro pre-maturation treatments (Pre-IVM), aimed to improve the developmental capability of immature oocytes, might behave differently depending on the oocyte metabolic status, when it is isolated from follicle [Luciano et al., 2011]. This study aims at identifying correlations between cumulus-oocyte complex (COC) morphology and oocyte chromatin configuration and secondly at testing the hypothesis that only fully grown oocytes at earlier stages of differentiation with loosely compacted chromatin (GV1) can benefit from Pre-IVM treatment. COCs were collected from bovine 2-6mm ovarian follicles, and further divided in three groups according to their morphology (Class-1, 2 and 3) as previously described [Blondin & Sirard, 1995]. Analysis of chromatin configuration revealed that only Class-1 COC was enriched in GV1 oocyte, while Class-2 and 3 presented a similar distribution of GV1, GV2 and GV3 oocytes, where GV2 and 3 oocytes are characterized by increased chromatin compaction [Lodde et al., 2007]. Then COCs were divided into two groups, one containing Class-1 COCs and the other containing Class-2 and 3 COCs and subjected to pre-IVM for 6 hours in presence of cilostamide and 10-4 UI/ml rhFSH. Finally, COCs underwent standard in vitro maturation (IVM) for 22 hours, in vitro fertilization and embryo culture. Blastocyst rate and embryos cell number were assessed at day 7. Pre-IVM positively affected developmental competences of Class-1, while in Classes 2 and 3 Pre-IVM had detrimental effects.In conclusion COCs morphology could be used as a non-invasive approach to select population of oocyte with different cultural needs. These data could be useful in setting-up dedicated IVM protocols considering specific genes and pathways to improve IVP efficiency.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91405203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. Catozzi, C. Lecchi, C. Gini, F. Ceciliani, R. Finotello, L. Ressel
MicroRNA (miRNA), a class of small, non-coding RNA - regulating post-transcriptionally protein expression - are emerging as clinical biomarkers in many pathologies, including cancer [1]. Since miRNA are supposed to represent fundamental key regulators, better understanding of melanoma biology is essential to improve staging and therapy. The aim of the study was to investigate whether miRNA expression can vary in canine melanoma samples derived from formalin-fixed-paraffin-embedded (FFPE) tissues. Experimental design of the study included three groups, each one composed of 7 animals: i) control healthy skin group ii) oral melanoma group iii) skin melanoma group. The histhopatology and immunoistochemistry details of dogs included in the study are previously reported [2]. Two tissue slides were used for miRNA extraction. The expression levels of seven miRNA - miR-145, miR-146a, miR-425-5p, miR-223, miR-365, miR-155 and miR-134 - were detected and assessed by qPCR using TaqMan® probes [3-5]. Five miRNA were significantly up-(n=3) or down-(n=2) regulated. In details, miR-146a and miR-155 abundance was increased as compared with control in both oral and skin melanoma (Fig 1 B,E) ( p = 0.004 and 0.014 and p = 0.043 and 0.035 respectively), while the levels of miR-145 and miR-365 were lower (Fig 1 A,D) ( p = 0.018 and 0.008 and p = 0.01 and 0.028, respectively). MiR-425-5p was upregulated ( p = 0.039) only in skin melanoma (Fig. 1 C). Furthermore, functional analysis, carried out using miRNet web-based tool, showed that 76 genes related to cancer-associated pathways were possible target of these five microRNA ( p = 6.95E-9); in particular, 21 target genes were associated with melanoma ( p = 1.47E-5), including BRAF and CDK, E2F, FGF and PIK3 families. In conclusion, miR-145, miR-146a, miR-425-5p, miR-365 and miR-155 are differentially expressed in melanoma and healthy FFPE samples, suggesting that they may play a role in canine melanoma pathogenesis and/or progression.
{"title":"Expression Analysis of MicroRNAs in FFPE samples of canine cutaneous and oral melanoma by RT-qPCR","authors":"C. Catozzi, C. Lecchi, C. Gini, F. Ceciliani, R. Finotello, L. Ressel","doi":"10.13130/2283-3927/8374","DOIUrl":"https://doi.org/10.13130/2283-3927/8374","url":null,"abstract":"MicroRNA (miRNA), a class of small, non-coding RNA - regulating post-transcriptionally protein expression - are emerging as clinical biomarkers in many pathologies, including cancer [1]. Since miRNA are supposed to represent fundamental key regulators, better understanding of melanoma biology is essential to improve staging and therapy. The aim of the study was to investigate whether miRNA expression can vary in canine melanoma samples derived from formalin-fixed-paraffin-embedded (FFPE) tissues. Experimental design of the study included three groups, each one composed of 7 animals: i) control healthy skin group ii) oral melanoma group iii) skin melanoma group. The histhopatology and immunoistochemistry details of dogs included in the study are previously reported [2]. Two tissue slides were used for miRNA extraction. The expression levels of seven miRNA - miR-145, miR-146a, miR-425-5p, miR-223, miR-365, miR-155 and miR-134 - were detected and assessed by qPCR using TaqMan® probes [3-5]. Five miRNA were significantly up-(n=3) or down-(n=2) regulated. In details, miR-146a and miR-155 abundance was increased as compared with control in both oral and skin melanoma (Fig 1 B,E) ( p = 0.004 and 0.014 and p = 0.043 and 0.035 respectively), while the levels of miR-145 and miR-365 were lower (Fig 1 A,D) ( p = 0.018 and 0.008 and p = 0.01 and 0.028, respectively). MiR-425-5p was upregulated ( p = 0.039) only in skin melanoma (Fig. 1 C). Furthermore, functional analysis, carried out using miRNet web-based tool, showed that 76 genes related to cancer-associated pathways were possible target of these five microRNA ( p = 6.95E-9); in particular, 21 target genes were associated with melanoma ( p = 1.47E-5), including BRAF and CDK, E2F, FGF and PIK3 families. In conclusion, miR-145, miR-146a, miR-425-5p, miR-365 and miR-155 are differentially expressed in melanoma and healthy FFPE samples, suggesting that they may play a role in canine melanoma pathogenesis and/or progression.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75163019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. D'urso, S. Faverzani, G. Barella, F. D. Cesare, D. Gioeni, V. Rabbogliatti, D. Stefanello, G. Ravasio
Several methods are described in veterinary medicine to perform and assess correct epidural needle placement to provide effective epidural analgesia (Adami et al 2017). The aim of this study is to evaluate the feasibility of an ultrasound longitudinal sagittal approach to epidural catheter placement using a biopsy needle guide. Seven dog cadavers were used in the study. With the cadaver in sternal recumbence, a 5-8 MHz microconvex transducer provided with a 16-gauge biopsy guide was positioned to obtain a longitudinal sagittal scan of the spinal process of L7 and the sacral crest; the epidural space was identified between two parallel hyperechoic lines and, as the trajectory of the biopsy guide crossed them, a 17G Tuohy needle was used to insert a 19G epidural catheter. Correct catheter placement was visualised through a resection of the column between L2 and L3. Firstly, an expert echographist (operator C1) visualised the ultrasonographic landmarks, while catheter placement was performed by an expert anaesthetist (operator A), a student (operator B) and another expert echographist (operator C2) (double-operator technique); secondly, operator A and C2 performed alone the whole procedure (single-operator technique); lastly all operators performed a blind procedure (Jones 2001). Operator A failed 2/7 single-operator procedures; time to perform the blind technique was statistically lower than the double-operator technique (75 ± 132.4 vs 91.6 ± 79.3 seconds). Operator C2 failed 3/7 blind procedures, scoring the higher total time of performance (329.3 ± 271.2 seconds), but was able to perform both the double- and single-operator technique without significant difference with operator A, despite a faster time in positioning the probe. Operator B showed a higher repositioning attempts of the needle with the double-operator procedure compared to the blind one. Ultrasound guidance appears to be a promising technique to ease catheter placement also by operators inexperienced of locoregional techniques.
兽医学中描述了几种方法来执行和评估正确的硬膜外针头放置，以提供有效的硬膜外镇痛(Adami等，2017)。本研究的目的是评估超声纵向矢状入路硬膜外导管放置活检针引导的可行性。研究中使用了7具狗的尸体。当尸体处于胸骨平躺状态时，放置一个5-8 MHz微凸换能器，并配备一个16号活检指南，以获得L7脊柱突和骶骨的纵向矢状面扫描;在两条平行的高回声线之间确定硬膜外间隙，当活检指南的轨迹与它们相交时，使用17G Tuohy针插入19G硬膜外导管。通过切除L2和L3之间的柱可见导管正确放置。首先，超声专家(操作员C1)可视化超声地标，导管放置由麻醉专家(操作员A)，学生(操作员B)和另一名超声专家(操作员C2)(双操作员技术)完成;第二，操作员A和C2单独完成整个过程(单操作员技术);最后，所有作业者都进行了盲操作(Jones 2001)。操作员A失败2/7单操作员程序;采用盲法的时间(75±132.4 vs 91.6±79.3)在统计学上低于双操作法。操作人员C2在3/7盲操作中失败，总执行时间更高(329.3±271.2秒)，但能够执行双操作和单操作技术，与操作人员A没有显著差异，尽管定位探针的时间更快。与盲人手术相比，双操作者手术B显示出更高的针头重新定位尝试。超声引导似乎是一种很有前途的技术，以减轻导管放置，即使操作人员缺乏局部技术。
{"title":"Ultrasound-guided epidural catheter placement with a new technique: preliminary cadaveric study.","authors":"E. D'urso, S. Faverzani, G. Barella, F. D. Cesare, D. Gioeni, V. Rabbogliatti, D. Stefanello, G. Ravasio","doi":"10.13130/2283-3927/8389","DOIUrl":"https://doi.org/10.13130/2283-3927/8389","url":null,"abstract":"Several methods are described in veterinary medicine to perform and assess correct epidural needle placement to provide effective epidural analgesia (Adami et al 2017). The aim of this study is to evaluate the feasibility of an ultrasound longitudinal sagittal approach to epidural catheter placement using a biopsy needle guide. Seven dog cadavers were used in the study. With the cadaver in sternal recumbence, a 5-8 MHz microconvex transducer provided with a 16-gauge biopsy guide was positioned to obtain a longitudinal sagittal scan of the spinal process of L7 and the sacral crest; the epidural space was identified between two parallel hyperechoic lines and, as the trajectory of the biopsy guide crossed them, a 17G Tuohy needle was used to insert a 19G epidural catheter. Correct catheter placement was visualised through a resection of the column between L2 and L3. Firstly, an expert echographist (operator C1) visualised the ultrasonographic landmarks, while catheter placement was performed by an expert anaesthetist (operator A), a student (operator B) and another expert echographist (operator C2) (double-operator technique); secondly, operator A and C2 performed alone the whole procedure (single-operator technique); lastly all operators performed a blind procedure (Jones 2001). Operator A failed 2/7 single-operator procedures; time to perform the blind technique was statistically lower than the double-operator technique (75 ± 132.4 vs 91.6 ± 79.3 seconds). Operator C2 failed 3/7 blind procedures, scoring the higher total time of performance (329.3 ± 271.2 seconds), but was able to perform both the double- and single-operator technique without significant difference with operator A, despite a faster time in positioning the probe. Operator B showed a higher repositioning attempts of the needle with the double-operator procedure compared to the blind one. Ultrasound guidance appears to be a promising technique to ease catheter placement also by operators inexperienced of locoregional techniques.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74573574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
B. Bolis, M. Schrank, J. Fusi, A. Mollo, M. Veronesi
The full knowledge of the normal fetal fluids composition could be useful in the dog for the better management of newborns. The aim of the present study was to define the biochemical composition of amniotic fluid of puppies born by elective Caesarean section (CS) at term of pregnancy. The study enrolled 24 purebred bitches, classified into small size ( 20kg). All the bitches were healthy and clinically monitored from mating until parturition. For all the bitches an elective CS at term of pregnancy was performed [1]. For each puppy, the amniotic fluid was collected, immediately centrifuged and frozen at – 20° C until analysis for ALB, AMY, TB, CHOL, CK, ALP, GGT, AST, ALT, LDH, Mg, Ca, K, Na, Trig, BUN, Glc, TP, CREA, LIP, Cl, and GLOB. Data were analyzed by ANCOVA to verify the possible effects of parity, breed body size and newborn gender on amniotic biochemical composition. A total of 69 amniotic fluid samples were collected. The amniotic mean±SD and min-max values for each parameter were defined. LDH (p<0.01) and CK activity (p<0.05), as well as Glc concentrations (p<0.0001) were negatively influenced by the parity. AMY activity was significantly (p<0.05) higher in large sized (44.2±20.87 U/L) respect to small sized dogs (30.3±19.89 U/L), while lower (p<0.05) CHOL amniotic concentrations were found in small sized (3.0±2.71 mg/dl) as compared to large sized (3.9±2.93 mg/dl) dogs. Gender of the newborn did not influence the amniotic biochemical composition. The preliminary results of this study showed some similarities as well as some differences concerning the biochemical composition of the amniotic fluid in dogs at term of pregnancy if compared to data reported for the cat [2]. Furthermore, the results suggested that, in dogs, some amniotic parameters could be influenced by breed body size and by parity.
{"title":"Biochemical composition of amniotic fluid in normal puppies at term of pregnancy: preliminary data","authors":"B. Bolis, M. Schrank, J. Fusi, A. Mollo, M. Veronesi","doi":"10.13130/2283-3927/8343","DOIUrl":"https://doi.org/10.13130/2283-3927/8343","url":null,"abstract":"The full knowledge of the normal fetal fluids composition could be useful in the dog for the better management of newborns. The aim of the present study was to define the biochemical composition of amniotic fluid of puppies born by elective Caesarean section (CS) at term of pregnancy. The study enrolled 24 purebred bitches, classified into small size ( 20kg). All the bitches were healthy and clinically monitored from mating until parturition. For all the bitches an elective CS at term of pregnancy was performed [1]. For each puppy, the amniotic fluid was collected, immediately centrifuged and frozen at – 20° C until analysis for ALB, AMY, TB, CHOL, CK, ALP, GGT, AST, ALT, LDH, Mg, Ca, K, Na, Trig, BUN, Glc, TP, CREA, LIP, Cl, and GLOB. Data were analyzed by ANCOVA to verify the possible effects of parity, breed body size and newborn gender on amniotic biochemical composition. A total of 69 amniotic fluid samples were collected. The amniotic mean±SD and min-max values for each parameter were defined. LDH (p<0.01) and CK activity (p<0.05), as well as Glc concentrations (p<0.0001) were negatively influenced by the parity. AMY activity was significantly (p<0.05) higher in large sized (44.2±20.87 U/L) respect to small sized dogs (30.3±19.89 U/L), while lower (p<0.05) CHOL amniotic concentrations were found in small sized (3.0±2.71 mg/dl) as compared to large sized (3.9±2.93 mg/dl) dogs. Gender of the newborn did not influence the amniotic biochemical composition. The preliminary results of this study showed some similarities as well as some differences concerning the biochemical composition of the amniotic fluid in dogs at term of pregnancy if compared to data reported for the cat [2]. Furthermore, the results suggested that, in dogs, some amniotic parameters could be influenced by breed body size and by parity.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82920613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Staphylococcus pseudintermedius is a newly described species of Staphylococcus regarded as the main causative agent of canine pyoderma [1]. S. pseudintermedius infection was recently described in humans. An important feature of this pathogen is the high genetic identity with two other species of staphylococci, namely S. intermedius and S. delphini , which are included all together in the Staphylococcus Intermedius Group (SIG) [2]. This scenario seriously hampers phenotypic differentiation of these three pathogens. Despite this, only in 2008 was described the first molecular protocol for diagnostic identification of S. pseudintermedius [3]. The aim of this work was to investigate the presence of different biotypes of S. pseudintermedius obtained from clinically relevant cases of pyoderma in dogs using three molecular methods commonly used to type bacteria: the Ribosomal Spacers Amplification (RSA), the Random Amplification of Polymorphic DNA (RAPD) and the Restriction Fragment Length Polymorphism (RFLP). A total of 46 different strains were included in this work. The application of the RSA technique, which was applied here for the first time, identified the presence of S. pseudintermedius , although it did not allow any differentiation between biotypes. The RAPD assay showed a single cluster that assembles all the interested strains that are grouped in three different sub-clusters ( Fig. 1 ). The RFLP technique showed the most discriminative power, providing the opportunity to clearly identify this bacterium. In conclusion, the use of these three different techniques allows to clearly identify S. pseudintermedius and to observe the presence of different biotypes. In future it could be interesting to couple these results with the determination of the antibiotic resistance in order to verify if certain Multi Drug Resistant strains have particular RSA and RAPD profiles.
{"title":"Molecular typing of Staphylococcus pseudintermedius canine strains by three commonly used techniques","authors":"G. Meroni, S. Morandi, M. Brasca, P. Martino","doi":"10.13130/2283-3927/8375","DOIUrl":"https://doi.org/10.13130/2283-3927/8375","url":null,"abstract":"Staphylococcus pseudintermedius is a newly described species of Staphylococcus regarded as the main causative agent of canine pyoderma [1]. S. pseudintermedius infection was recently described in humans. An important feature of this pathogen is the high genetic identity with two other species of staphylococci, namely S. intermedius and S. delphini , which are included all together in the Staphylococcus Intermedius Group (SIG) [2]. This scenario seriously hampers phenotypic differentiation of these three pathogens. Despite this, only in 2008 was described the first molecular protocol for diagnostic identification of S. pseudintermedius [3]. The aim of this work was to investigate the presence of different biotypes of S. pseudintermedius obtained from clinically relevant cases of pyoderma in dogs using three molecular methods commonly used to type bacteria: the Ribosomal Spacers Amplification (RSA), the Random Amplification of Polymorphic DNA (RAPD) and the Restriction Fragment Length Polymorphism (RFLP). A total of 46 different strains were included in this work. The application of the RSA technique, which was applied here for the first time, identified the presence of S. pseudintermedius , although it did not allow any differentiation between biotypes. The RAPD assay showed a single cluster that assembles all the interested strains that are grouped in three different sub-clusters ( Fig. 1 ). The RFLP technique showed the most discriminative power, providing the opportunity to clearly identify this bacterium. In conclusion, the use of these three different techniques allows to clearly identify S. pseudintermedius and to observe the presence of different biotypes. In future it could be interesting to couple these results with the determination of the antibiotic resistance in order to verify if certain Multi Drug Resistant strains have particular RSA and RAPD profiles.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77706459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Chiapparini, F. Vizzarri, C. Corino, R. Rossi, G. Maiorano
In the recent years, research has focused the attention on the use of substances able to improve both animal health and meat quality. The objective of this study was to investigate the effect of dietary supplementation with natural extracts in rabbit does on reproductive parameters. Natural extracts have been widely reported to have potent antioxidant, anti-inflammatory and antimicrobic activities related especially to their phenolic content (Pereira et al., 2009). In rabbit production, does are normally inseminated 11 days after parturition, with kits being weaned at 35 days (Rebollar et al., 2006). The reproductive phase is a critical period because the animals have few days to reconstitute their body reserves (Castellini et al., 2003). Therefore, the interaction between nutrition and reproduction is crucial in this specie (Butler, 2000; Roche et al., 2000). The trial was performed at the Research Institute for animal production in Nitra (Slovak Republic). Sixty does were randomly selected, artificially inseminated and divided into three experimental groups. The first fed a basal diet (C), the second one (T1) received 0.3% of natural extracts mixture and the third one (T2) an integration of 0.6% of natural extracts mixture for gestation and lactation period (65 days) The animals were allocated in individual flat-deck cages with a closeable nest box. At parturition, the total number of kits born and stillborn and litters weights were recorded. The data were analyzed by one way Analysis of Variance (ANOVA) using SPSS (IBM.-SSPS Statistics 24). Dietary supplementation did not affect (P>0.05) number of kids born (8.0 ± 1.0 C vs 7.3 ± 0.97 T1 and 7.4 ± 1.0 T2) and birth weight (63 ± 2.0 g C vs 60.1 ± 2.3 g T1 and 61.0 ± 2.4 g T2). The administration of natural extract in does did not improve (P>0.05) the kits average daily gain (20.54 ± 1.3 g/d C vs 21.92 ± 0.5 g/d T1 and 20.93 ± 0.9 g/d T2) and body weight at weaning (829 ± 16.6 g C vs 834 ± 26.6 g T1 and 826 ± 26.8 g T2). These preliminary data showed that at the present dosage, the natural extracts mixture is not able to affect does reproductive performance. However, further research is needed to confirm the present data and explore the mechanism of action of this natural mixture.
近年来，研究将注意力集中在能够改善动物健康和肉质的物质的使用上。本研究旨在探讨饲粮中添加天然提取物对家兔生殖参数的影响。天然提取物已被广泛报道具有有效的抗氧化、抗炎和抗菌活性，特别是与它们的酚类含量有关(Pereira等人，2009)。在家兔生产中，公兔通常在分娩后11天进行授精，幼兔在35天断奶(rebolar等，2006)。繁殖阶段是一个关键时期，因为动物只有很少的时间来重建它们的身体储备(Castellini et al.， 2003)。因此，营养和繁殖之间的相互作用对这个物种至关重要(Butler, 2000;Roche et al.， 2000)。试验是在Nitra(斯洛伐克共和国)的动物生产研究所进行的。随机选取60只进行人工授精，分为3个实验组。在妊娠期和哺乳期(65 d)，第一组饲喂基础饲粮(C)，第二组(T1)饲喂0.3%的天然提取物混合物，第三组(T2)饲喂0.6%的天然提取物混合物。在分娩时，记录出生和死产的幼崽总数和窝重。采用SPSS (IBM)软件进行单因素方差分析(ANOVA)。——ssps统计饲粮添加量对出生仔数(8.0±1.0 C vs 7.3±0.97 T1和7.4±1.0 T2)和出生体重(63±2.0 g C vs 60.1±2.3 g T1和61.0±2.4 g T2)无显著影响(P>0.05)。饲粮中添加天然提取物对断奶仔猪平均日增重(20.54±1.3 g/d C vs 21.92±0.5 g/d T1和20.93±0.9 g/d T2)和断奶体重(829±16.6 g C vs 834±26.6 g T1和826±26.8 g T2)均无显著影响(P>0.05)。这些初步数据表明，在目前的剂量下，天然提取物混合物不会影响其繁殖性能。然而，需要进一步的研究来证实目前的数据，并探索这种天然混合物的作用机制。
{"title":"Effect of dietary natural extracts mixture on rabbit does reproductive performances: preliminary data.","authors":"S. Chiapparini, F. Vizzarri, C. Corino, R. Rossi, G. Maiorano","doi":"10.13130/2283-3927/8365","DOIUrl":"https://doi.org/10.13130/2283-3927/8365","url":null,"abstract":"In the recent years, research has focused the attention on the use of substances able to improve both animal health and meat quality. The objective of this study was to investigate the effect of dietary supplementation with natural extracts in rabbit does on reproductive parameters. Natural extracts have been widely reported to have potent antioxidant, anti-inflammatory and antimicrobic activities related especially to their phenolic content (Pereira et al., 2009). In rabbit production, does are normally inseminated 11 days after parturition, with kits being weaned at 35 days (Rebollar et al., 2006). The reproductive phase is a critical period because the animals have few days to reconstitute their body reserves (Castellini et al., 2003). Therefore, the interaction between nutrition and reproduction is crucial in this specie (Butler, 2000; Roche et al., 2000). The trial was performed at the Research Institute for animal production in Nitra (Slovak Republic). Sixty does were randomly selected, artificially inseminated and divided into three experimental groups. The first fed a basal diet (C), the second one (T1) received 0.3% of natural extracts mixture and the third one (T2) an integration of 0.6% of natural extracts mixture for gestation and lactation period (65 days) The animals were allocated in individual flat-deck cages with a closeable nest box. At parturition, the total number of kits born and stillborn and litters weights were recorded. The data were analyzed by one way Analysis of Variance (ANOVA) using SPSS (IBM.-SSPS Statistics 24). Dietary supplementation did not affect (P>0.05) number of kids born (8.0 ± 1.0 C vs 7.3 ± 0.97 T1 and 7.4 ± 1.0 T2) and birth weight (63 ± 2.0 g C vs 60.1 ± 2.3 g T1 and 61.0 ± 2.4 g T2). The administration of natural extract in does did not improve (P>0.05) the kits average daily gain (20.54 ± 1.3 g/d C vs 21.92 ± 0.5 g/d T1 and 20.93 ± 0.9 g/d T2) and body weight at weaning (829 ± 16.6 g C vs 834 ± 26.6 g T1 and 826 ± 26.8 g T2). These preliminary data showed that at the present dosage, the natural extracts mixture is not able to affect does reproductive performance. However, further research is needed to confirm the present data and explore the mechanism of action of this natural mixture.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79941298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lake Garda is located in Insubria region, that is known for being the most populated and industrialized area of Italy (Camusso et al., 2001). Therefore, the Lake water, and also the fish species present, could be affected by environmental contamination. European eel ( Anguilla anguilla ) are considered as suitable matrix for biomonitoring environmental contaminants in European water (Belpaire et al., 2007), being widespread in many European waters and highly contaminated by lipophilic compounds, due to the high lipid content (up to 40%) (Larsson et al., 1991). Moreover, eel is an edible species (its farming currently supplies approximately 45,000 tons/year) (Nielsen et al., 2008), so it also represents a public health issue. Based on these considerations, the aim of this study was to evaluate the occurrence of fourteen organochlorine pesticides (OCs) in forty-five eels ( Anguilla anguilla ) from Lake Garda, using Accelerated Solvent Extraction (ASE) procedure for the analytes extraction and Gas chromatography coupled with Tandem Mass Spectrometry (GC-MS/MS) for the analysis of OCs. GC-MS/MS analysis was developed and validated according to the SANTE/11945/2015 guidelines. Uncontaminated eel sample (previously checked for the presence of OCs and considered blank with a concentration of compounds < Limit of Detection) were used for all procedure's optimization steps. For all the OCs analysed, satisfactory results were achieved. Regarding eel samples, several pesticides were detected, but DDTs (DDT and its metabolites) were found with the highest prevalence (92 %). The concentration rage was from not detected (n.d.) to 19000 ng g -1 . Although DDTs levels in the environment are declining (Albaiges et al., 2011), they continue to bioaccumulate in tissues of human and animal and biomagnify in food chains.
加尔达湖位于Insubria地区，该地区以意大利人口最多、工业化程度最高而闻名(Camusso et al.， 2001)。因此，湖水和鱼类都可能受到环境污染的影响。欧洲鳗鲡(Anguilla Anguilla)被认为是欧洲水域生物监测环境污染物的合适基质(Belpaire等，2007)，由于其高脂含量(高达40%)，在许多欧洲水域广泛存在，并受到亲脂化合物的严重污染(Larsson等，1991)。此外，鳗鱼是一种可食用物种(目前其养殖供应量约为45,000吨/年)(Nielsen et al.， 2008)，因此它也代表了一个公共卫生问题。基于上述考虑，本研究采用加速溶剂萃取法(ASE)提取鳗鲡(Anguilla Anguilla) 45只鳗鲡中14种有机氯农药(OCs)的含量，气相色谱-串联质谱联用(GC-MS/MS)对OCs进行分析。GC-MS/MS分析是根据SANTE/11945/2015指南开发和验证的。所有程序的优化步骤都使用未受污染的鳗鱼样本(先前检查了OCs的存在，并且认为化合物浓度<检测限为空白)。所有的OCs分析都取得了令人满意的结果。在鳗鱼样本中检测到几种农药，但DDT(滴滴涕及其代谢物)的流行率最高(92%)。浓度范围从未检出(n.d)到19000 ng g -1。尽管环境中的ddt水平正在下降(Albaiges et al.， 2011)，但它们继续在人类和动物组织中进行生物积累，并在食物链中进行生物放大。
{"title":"Analyses of organochlorine pesticides residues in eels (Anguilla anguilla) from Lake Garda using Gas chromatography coupled with Tandem Mass Spectrometry (GC-MS/MS).","authors":"G. Labella, L. Chiesa, S. Panseri, F. Arioli","doi":"10.13130/2283-3927/8377","DOIUrl":"https://doi.org/10.13130/2283-3927/8377","url":null,"abstract":"Lake Garda is located in Insubria region, that is known for being the most populated and industrialized area of Italy (Camusso et al., 2001). Therefore, the Lake water, and also the fish species present, could be affected by environmental contamination. European eel ( Anguilla anguilla ) are considered as suitable matrix for biomonitoring environmental contaminants in European water (Belpaire et al., 2007), being widespread in many European waters and highly contaminated by lipophilic compounds, due to the high lipid content (up to 40%) (Larsson et al., 1991). Moreover, eel is an edible species (its farming currently supplies approximately 45,000 tons/year) (Nielsen et al., 2008), so it also represents a public health issue. Based on these considerations, the aim of this study was to evaluate the occurrence of fourteen organochlorine pesticides (OCs) in forty-five eels ( Anguilla anguilla ) from Lake Garda, using Accelerated Solvent Extraction (ASE) procedure for the analytes extraction and Gas chromatography coupled with Tandem Mass Spectrometry (GC-MS/MS) for the analysis of OCs. GC-MS/MS analysis was developed and validated according to the SANTE/11945/2015 guidelines. Uncontaminated eel sample (previously checked for the presence of OCs and considered blank with a concentration of compounds < Limit of Detection) were used for all procedure's optimization steps. For all the OCs analysed, satisfactory results were achieved. Regarding eel samples, several pesticides were detected, but DDTs (DDT and its metabolites) were found with the highest prevalence (92 %). The concentration rage was from not detected (n.d.) to 19000 ng g -1 . Although DDTs levels in the environment are declining (Albaiges et al., 2011), they continue to bioaccumulate in tissues of human and animal and biomagnify in food chains.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84140261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Perfluoroalkyl substances (PFASs) are a large class of fluorinated aliphatic chemical of anthropogenic origin with high chemical stability even at high temperatures and in presence of alkalis, strong acids or oxidizing agents (Lau et al. 2004). All these characteristics make them no biodegradable and very persistent in the environment, associated with adverse health risks (Eriksen et al. 2010). Food, especially fish and other seafood, is considered the main source of exposure to PFASs (EFSA 2012). In this preliminary study we developed and validated a sensitive, selective and specific method by LC-HRMS Orbitrap to monitor the presence of 16 PFASs in eel ( Anguilla Anguilla ) samples. The clean-up of the lyophilized samples consisted of a previous extraction step with acetonitrile to precipitate also proteins, followed by a purification step through Oasis® WAX SPE (Weak Anionic Exchange Solid Phase Extraction) cartridges. The method applied to 45 farmed eel samples from Lake Garda showed the presence of several PFASs, up to 10 in the same eel, in the order of ng/g (Fig.1). The results provided a representative situation of the PFASs contamination level of the lake, lower than those of others European countries (Hoff et al. 2005, Kwadijk et al. 2010).
全氟烷基物质(PFASs)是一类人为来源的氟化脂肪族化学品，即使在高温和碱、强酸或氧化剂存在下也具有很高的化学稳定性(Lau等人，2004年)。所有这些特征使它们无法生物降解，并且在环境中非常持久，具有不利的健康风险(Eriksen等人，2010年)。食物，特别是鱼类和其他海产品，被认为是全氟辛烷磺酸的主要暴露来源(EFSA 2012)。在本初步研究中，我们利用LC-HRMS Orbitrap建立并验证了一种灵敏、选择性和特异性的方法来监测鳗鲡(Anguilla Anguilla)样品中16种PFASs的存在。冻干样品的清理包括前一步用乙腈提取蛋白质，然后通过Oasis®WAX SPE(弱阴离子交换固相萃取)净化步骤。该方法应用于加尔达湖的45条养殖鳗鱼样本，结果显示存在几种PFASs，同一条鳗鱼中多达10种，以ng/g为数量级(图1)。结果提供了湖泊全氟磺酸污染水平的代表性情况，低于其他欧洲国家(Hoff et al. 2005, Kwadijk et al. 2010)。
{"title":"Preliminary study about the detection of perfluoroalkyl substances in eel samples of Lake Garda by liquid chromatography tandem mass high resolution mass spectrometry (LC-HRMS).","authors":"M. Nobile, S. Panseri, F. Arioli, L. Chiesa","doi":"10.13130/2283-3927/8376","DOIUrl":"https://doi.org/10.13130/2283-3927/8376","url":null,"abstract":"Perfluoroalkyl substances (PFASs) are a large class of fluorinated aliphatic chemical of anthropogenic origin with high chemical stability even at high temperatures and in presence of alkalis, strong acids or oxidizing agents (Lau et al. 2004). All these characteristics make them no biodegradable and very persistent in the environment, associated with adverse health risks (Eriksen et al. 2010). Food, especially fish and other seafood, is considered the main source of exposure to PFASs (EFSA 2012). In this preliminary study we developed and validated a sensitive, selective and specific method by LC-HRMS Orbitrap to monitor the presence of 16 PFASs in eel ( Anguilla Anguilla ) samples. The clean-up of the lyophilized samples consisted of a previous extraction step with acetonitrile to precipitate also proteins, followed by a purification step through Oasis® WAX SPE (Weak Anionic Exchange Solid Phase Extraction) cartridges. The method applied to 45 farmed eel samples from Lake Garda showed the presence of several PFASs, up to 10 in the same eel, in the order of ng/g (Fig.1). The results provided a representative situation of the PFASs contamination level of the lake, lower than those of others European countries (Hoff et al. 2005, Kwadijk et al. 2010).","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90292224","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. Curone, S. Panseri, C. Colombani, L. Chiesa, J. Filipe, M. Faustini
In the past decades, milk has been considered a mere supplier of nutrients, although its importance was considered paramount for the development and growth of newborns, a number of aspects regarding the biological functions of milk are still unknown. Several positive functional properties of milk derive from fatty acids (FA), mainly unsaturated fatty acids (UFA), either monounsaturated (MUFA) or polyunsaturated (PUFA) fatty acids. In particular, UFAs are considered functional components of food because of their positive effects on disease prevention (FAO, 2010; Connor W.E. ,2000; Wijendran V., 2004). The objective of this study was to characterize the fatty acid profile, the desaturase index and the atherogenic index in milk of local Italian bovine breeds (Cabannina, Varzese and Valdostana) and in a cosmopolitan breed (Holstein Friesian) during the first period of lactation. A total number of 129 cows have been enrolled (Friesian n=30, Cabannina n=30, Varzese n=30, Valdostana n=39) from three dairy farms with similar management and feeding conditions. Animals were chosen in order to have three classes of lactation stage: milk collections were carried out starting from 40±10 days (group A), 70±10 days (group B), and 130±10 days (group C). Milk samples have been analyzed by gas chromatography to obtain the fatty acid profile, on the basis of these results, the Desaturase and Atherogenic Indices were calculated. A number of differences between breeds have been found, in particular local breeds showed an higher percentages of UFA, MUFA, PUFA, and a higher UFA/SFA ratio, as well as lower desaturase indices (related to C14, C16 and C18) and atherogenic index, when compared to Friesian cows. The results can add further information aiming to re-evaluate an almost lost local treasure in Northern Italy.
{"title":"Fatty acid profile, Desaturase and Atherogenic indices in milk of Holstein Friesian and Italian autochthonous cattle breeds","authors":"G. Curone, S. Panseri, C. Colombani, L. Chiesa, J. Filipe, M. Faustini","doi":"10.13130/2283-3927/8403","DOIUrl":"https://doi.org/10.13130/2283-3927/8403","url":null,"abstract":"In the past decades, milk has been considered a mere supplier of nutrients, although its importance was considered paramount for the development and growth of newborns, a number of aspects regarding the biological functions of milk are still unknown. Several positive functional properties of milk derive from fatty acids (FA), mainly unsaturated fatty acids (UFA), either monounsaturated (MUFA) or polyunsaturated (PUFA) fatty acids. In particular, UFAs are considered functional components of food because of their positive effects on disease prevention (FAO, 2010; Connor W.E. ,2000; Wijendran V., 2004). The objective of this study was to characterize the fatty acid profile, the desaturase index and the atherogenic index in milk of local Italian bovine breeds (Cabannina, Varzese and Valdostana) and in a cosmopolitan breed (Holstein Friesian) during the first period of lactation. A total number of 129 cows have been enrolled (Friesian n=30, Cabannina n=30, Varzese n=30, Valdostana n=39) from three dairy farms with similar management and feeding conditions. Animals were chosen in order to have three classes of lactation stage: milk collections were carried out starting from 40±10 days (group A), 70±10 days (group B), and 130±10 days (group C). Milk samples have been analyzed by gas chromatography to obtain the fatty acid profile, on the basis of these results, the Desaturase and Atherogenic Indices were calculated. A number of differences between breeds have been found, in particular local breeds showed an higher percentages of UFA, MUFA, PUFA, and a higher UFA/SFA ratio, as well as lower desaturase indices (related to C14, C16 and C18) and atherogenic index, when compared to Friesian cows. The results can add further information aiming to re-evaluate an almost lost local treasure in Northern Italy.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76125395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Meazzi, S. Lauzi, A. Stranieri, S. Paltrinieri, A. Giordano
Feline Infectious Peritonitis (FIP) develops from a mutation of enteric feline coronaviruses (FCoVs) and an imbalance of the host immune response. The wide polymorphism of FCoVs is associated with the viral replication rate (Licitra et al. 2013) . Changes in the composition of the gut microbiota may induce quali-quantitative modifications in FCoVs and/or different immune profiles (Weese et al., 2015). Few information is available on feline gut microbiome and the association between microbiota and the predisposition to pathological conditions (Ramadan et al., 2014). The aim of this study is to provide preliminary data about the composition of gut microbiota in healthy cats compared with FCoV infected cats (with and without FIP), in order to evaluate whether changes of gut microbiota may induce changes in FCoV, in its genetic polymorphism and in the mucosal immunity. Screening analyses have been performed on 22 cats: - Routine hematology and biochemistry on EDTA and serum (included electrophoresis and alpha-1-acid glycoprotein measurement for cats suspected with FIP) - Nested RT-PCR-3’UTR on frozen faeces - Effusion evaluation - FIV/FeLV serology Due to strict inclusion criteria (cats younger than 2.5 years old, indoor and not assuming antibiotics in the previous two months) and based on the results obtained from the complete set of analysis, only 15 cats, specifically 5 cats for each of the following 3 groups: FIP- affected, healthy negative and positive for FCoV, have been recruited to perform the following analyses: - microbiota analysis through NGS of 16S rRNA gene (V4 region) amplicons followed by bioinformatic analysis - evaluation of secretory IgA (ELISA kit) - phylogenetic analysis of FCoVs S gene sequences Innovative results will be provided on the feline gut microbiota composition. These will be correlated with the presence and genetic polymorphisms of FCoV and mucosal defenses to establish significant correlations between the analysed factors.
猫传染性腹膜炎(FIP)是由肠道猫冠状病毒(fcov)突变和宿主免疫反应不平衡引起的。fcov的广泛多态性与病毒复制率有关(Licitra et al. 2013)。肠道微生物群组成的变化可能导致fcov和/或不同免疫谱的质-定量改变(Weese等，2015)。关于猫肠道微生物群以及微生物群与病理状况易感性之间的关系的信息很少(Ramadan et al.， 2014)。本研究的目的是提供健康猫与感染FCoV的猫(有和没有FIP)肠道微生物群组成的初步数据，以评估肠道微生物群的变化是否会引起FCoV的变化，其遗传多态性和粘膜免疫。对22只猫进行了筛选分析:-常规血液学和生化检测EDTA和血清(包括对疑似FIP的猫进行电泳和α -1-酸糖蛋白检测)-冷冻粪便的嵌套RT-PCR-3'UTR -积液评估- FIV/FeLV血清学由于严格的纳入标准(小于2.5岁、室内且前两个月未使用抗生素的猫)，基于完整的分析结果，只有15只猫，具体来说，我们招募了5只猫，分别用于以下3组:受FIP影响、健康、FCoV阴性和阳性的猫，进行以下分析:-通过16S rRNA基因(V4区)扩增子的NGS分析进行微生物群分析，然后进行生物信息学分析-分泌IgA (ELISA试剂盒)评估- FCoV S基因序列的系统发育分析将提供关于猫肠道微生物群组成的创新结果。这些将与FCoV和粘膜防御的存在和遗传多态性相关，以确定所分析因素之间的显著相关性。
{"title":"The gut microbiome and mucosal defenses in cats with coronaviruses: a pilot study","authors":"S. Meazzi, S. Lauzi, A. Stranieri, S. Paltrinieri, A. Giordano","doi":"10.13130/2283-3927/8410","DOIUrl":"https://doi.org/10.13130/2283-3927/8410","url":null,"abstract":"Feline Infectious Peritonitis (FIP) develops from a mutation of enteric feline coronaviruses (FCoVs) and an imbalance of the host immune response. The wide polymorphism of FCoVs is associated with the viral replication rate (Licitra et al. 2013) . Changes in the composition of the gut microbiota may induce quali-quantitative modifications in FCoVs and/or different immune profiles (Weese et al., 2015). Few information is available on feline gut microbiome and the association between microbiota and the predisposition to pathological conditions (Ramadan et al., 2014). The aim of this study is to provide preliminary data about the composition of gut microbiota in healthy cats compared with FCoV infected cats (with and without FIP), in order to evaluate whether changes of gut microbiota may induce changes in FCoV, in its genetic polymorphism and in the mucosal immunity. Screening analyses have been performed on 22 cats: - Routine hematology and biochemistry on EDTA and serum (included electrophoresis and alpha-1-acid glycoprotein measurement for cats suspected with FIP) - Nested RT-PCR-3’UTR on frozen faeces - Effusion evaluation - FIV/FeLV serology Due to strict inclusion criteria (cats younger than 2.5 years old, indoor and not assuming antibiotics in the previous two months) and based on the results obtained from the complete set of analysis, only 15 cats, specifically 5 cats for each of the following 3 groups: FIP- affected, healthy negative and positive for FCoV, have been recruited to perform the following analyses: - microbiota analysis through NGS of 16S rRNA gene (V4 region) amplicons followed by bioinformatic analysis - evaluation of secretory IgA (ELISA kit) - phylogenetic analysis of FCoVs S gene sequences Innovative results will be provided on the feline gut microbiota composition. These will be correlated with the presence and genetic polymorphisms of FCoV and mucosal defenses to establish significant correlations between the analysed factors.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85181226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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