Porcine Origin Research Articles

Widespread Dissemination of Plasmid-Mediated Tigecycline Resistance Gene tet(X4) in Enterobacterales of Porcine Origin.

ABSTRACTThe emergence of the plasmid-mediated high levels of the tigecycline resistance gene has drawn worldwide attention and has posed a major threat to public health. In this study, we investigated the prevalence of the tet(X4)-positive Enterobacterales isolates collected from a pig slaughterhouse and farms. A total of 101 tigecycline resistance strains were isolated from 353 samples via a medium with tigecycline, of which 33 carried tet(X4) (9.35%, 33/353) and 2 carried tet(X6) (0.57%, 2/353). These strains belong to seven different species, with Escherichia coli being the main host bacteria. Importantly, this report is the first one to demonstrate that tet(X4) was observed in Morganella morganii. Whole-genome sequencing results revealed that tet(X4)-positive bacteria can coexist with other resistance genes, such as blaNDM-1 and cfr. Additionally, we were the first to report that tet(X4) and blaNDM-1 coexist in a Klebsiella quasipneumoniae strain. The phylogenetic tree of 533 tet(X4)-positive E. coli strains was constructed using 509 strains from the NCBI genome assembly database and 24 strains from this study, which arose from 8 sources and belonged to 135 sequence types (STs) worldwide. We used Nanopore sequencing to interpret the selected 21 nonclonal and representative strains and observed that 19 tet(X4)-harboring plasmids were classified into 8 replicon types, and 2 tet(X6) genes were located on integrating conjugative elements. A total of 68.42% of plasmids carrying tet(X4) were transferred successfully with a conjugation frequency of 10−2 to 10−7. These findings highlight that diverse plasmids drive the widespread dissemination of the tigecycline resistance gene tet(X4) in Enterobacterales of porcine origin.IMPORTANCE Tigecycline is considered to be the last resort of defense against diseases caused by broad-spectrum resistant Gram-negative bacteria. In this study, we systematically analyzed the prevalence and genetic environments of the resistance gene tet(X4) in a pig slaughterhouse and farms and the evolutionary relationship of 533 tet(X4)-positive Escherichia coli strains, including 509 tet(X4)-positive E. coli strains selected from the 27,802 assembled genomes of E. coli from the NCBI between 2002 and 2022. The drug resistance of tigecycline is widely prevalent in pig farms where tetracycline is used as a veterinary drug. This prevalence suggests that pigs are a large reservoir of tet(X4) and that tet(X4) can spread horizontally through the food chain via mobile genetic elements. Furthermore, tetracycline resistance may drive tigecycline resistance through some mechanisms. Therefore, it is important to monitor tigecycline resistance, develop effective control measures, and focus on tetracycline use in the pig farms.

Time-resolved characterization of the innate immune response in the respiratory epithelium of human, porcine, and bovine during influenza virus infection.

Viral cross-species transmission is recognized to be a major threat to both human and animal health, however detailed information on determinants underlying virus host tropism and susceptibility is missing. Influenza C and D viruses (ICV, IDV) are two respiratory viruses that share up to 50% genetic similarity, and both employ 9-O-acetylated sialic acids to enter a host cell. While ICV infections are mainly restricted to humans, IDV possesses a much broader host tropism and has shown to have a zoonotic potential. This suggests that additional virus–host interactions play an important role in the distinct host spectrum of ICV and IDV. In this study, we aimed to characterize the innate immune response of the respiratory epithelium of biologically relevant host species during influenza virus infection to identify possible determinants involved in viral cross-species transmission. To this end, we performed a detailed characterization of ICV and IDV infection in primary airway epithelial cell (AEC) cultures from human, porcine, and bovine origin. We monitored virus replication kinetics, cellular and host tropism, as well as the host transcriptional response over time at distinct ambient temperatures. We observed that both ICV and IDV predominantly infect ciliated cells, independently from host and temperature. Interestingly, temperature had a profound influence on ICV replication in both porcine and bovine AEC cultures, while IDV replicated efficiently irrespective of temperature and host. Detailed time-resolved transcriptome analysis revealed both species-specific and species uniform host responses and highlighted 34 innate immune-related genes with clear virus-specific and temperature-dependent profiles. These data provide the first comprehensive insights into important common and species-specific virus-host dynamics underlying the distinct host tropism of ICV and IDV, as well as possible determinants involved in viral cross-species transmission.

Superovulation of high-producing Holstein lactating dairy cows with human recombinant FSH and hMG

Superovulation of high-producing dairy cows is a challenging subject in dairy farms with respect to the cost, dose and type of gonadotropin. The objectives of this study were to compare three gonadotropin products: Folltropin-V® (highly purified FSH with porcine origin), Cinnal-f® (recombinant human FSH) and Menotropins® (hMG) for superovulation in high-producing Holstein lactating dairy cows and to investigate the pregnancy outcomes achieved following transferring embryos recovered from donors treated with different gonadotropins. Healthy high-producing Holstein lactating dairy cows (n = 30; milk production: 46.35 ± 8.78 kg; parity: 2–4; days in milk: 80–130 days) without any puerperal problems were selected as donors. On Day 10 after estrus (Day 0 of superovulation), donors (10 cows in each experimental groups) received Folltropin-V® (400 mg NIH, dissolved in 20 ml), Cinnal-f® (20 vials; each vial of 1 ml contains 75 IU Follitropin alfa) and Menotropins ® (20 ampules; each ampule of 1 ml contains 75 IU FSH and 75 IU LH), administered twice daily, in decreasing doses (4,4; 3,3; 2,2; 1,1 ml), over 4 days. On Day 2 of superovulation, donors received 3 doses of prostaglandin F2α analogue, 6 h apart. They were inseminated twice with a frozen semen at 12 and 24 h after standing estrus. Concurrent with the second insemination, donors received 2500 IU hCG (Karma Pharmatech GmbH, Germany). On Day 7 after standing estrus, superovulatory responses (number of CLs, total ova/embryos and transferable embryos) were recorded and Code 1 embryos, recovered from each treated donors, were transferred to synchronized heifers. Pregnancy was detected on Day 30 and 60 after AI. Gestation length, the number and weight of live births were recorded. Data were analyzed using Proc GLM, Proc Mixed and Proc Genmod of SAS. The respective number of corpora lutea, total number of ova/embryos and transferable embryos were not different among donors received Cinnal-f (25.5 ± 3.01, 11.2 ± 2.77, 5.1 ± 0.86), Menotropins (24.0 ± 3.21, 9.0 ± 2.04, 6.3 ± 1.74) and Folltropin-V (20.3 ± 3.21, 8.9 ± 1.90, 5.1 ± 1.16; P > 0.05). Pregnancy rates on Day 30 was similar among treatment groups (P > 0.05). However, pregnancy rates on Day 60 and the number of calves born healthy was less in heifers that received embryos from Cinnal-f treated donors (P < 0.05). In conclusion, Cinnal-f and Menotropins could provide similar superovulatory response to Folltropin-V for superovulation of high-producing Holstein lactating dairy cows.

Breast reconstruction with silicone prosthesis and acellular dermal matrix of porcine origin: Retrospective study of 84 cases

Breast reconstruction (BR) using a prosthesis implant (PI) associated with an acellular dermal matrix (ADM) is a known method that has been the subject of discussion in recent years. The objective of this study was to quantify the rate of PI removal after BR using prosthesis combined with ADM, and to identify the risk factors in the event of removal. This was a retrospective study conducted between 2010 and 2015. Inclusion criteria were patients who had undergone immediate (IBR) or delayed (DBR) breast reconstruction with placement of a PI associated with porcine ADM. The primary endpoint was the postoperative removal of the PI. In all, 84 reconstructions were performed. The mean age of the population was 57.5 years. 25.9% of the patients were active smokers at the time of surgery. 89.5% of patients had previously benefited from ipsilateral breast radiation therapy (IBRT) in DBR, 10.5% in IBR. The PI deposition rate, all BR combined, was 21.4%. It was 52.17% in smokers and 9.84% in non-smokers (P<0.0001), making smoking an independent risk factor for reconstruction failure (hazard ratio (HR)=7.4, 95%CI [2.64-20.9]). IBRT was also a risk factor for PI removal, especially when performed after IBR (HR=8.1, 95%CI [1.1-62.1]). Smoking and adjuvant IBRT are risk factors for failure of reconstruction by PI associated with ADM. This type of reconstruction should be selected for non-smokers who have not undergone IBRT and therefore could be a therapeutic alternative in the BR panel.

PSVI-13 Identifying Probiotic Bacteria to Reduce Pathogen Colonization in Nursery Pigs

Abstract Weaning stress and the negative health effects that occur with it cause serious concerns for pork producers. The abrupt environmental and dietary transitions that occur at weaning often take place before the gut microbiome is stabilized in the piglet. This leaves weanling piglets vulnerable to gut dysbiosis and increases susceptibility to enteric pathogens. The infections by opportunistic pathogens can be prevented with antibiotics, but due to growing concerns surrounding antimicrobial resistance in bacterial populations, direct-fed microbial (DFM) formulations have emerged as an alternative prevention method. In the current study, we isolated Lactobacillus spp. and Bifidobacterium spp. from fecal samples collected from neonatal pigs and sows that are capable of inhibiting colonization of Salmonella spp. and Escherichia coli isolated from sick pigs. Diluted fecal samples were plated on Bifidus selective medium (BSM) and deMan, Rogosa Sharpe (MRS) medium to isolate Bifidobacterium and Lactobacillus, respectively. Bacterial isolates were screened against Salmonella and E. coli isolates of porcine origin using a medium consisting of 50% MRS and 50% brain, heart, lung infusion (BHI) using disk diffusion assays. The resulting isolates that demonstrated the greaest degree of pathogen inhibition were characterized using 16s rRNA sequencing. Sequence analysis identified the top isolates as L. mucosae, L. reuteri, and B. thermacidophilum subsp. porcinum. Currently, genome sequencing of the selected isolates is underway to identify the mechanisms of inhibition.

Osteogenic differentiation of human dental pulp stem cells in decellularised adipose tissue solid foams.

3D cell culture systems based on biological scaffold materials obtainable from both animal and human tissues constitute very interesting tools for cell therapy and personalised medicine applications. The white adipose tissue (AT) extracellular matrix (ECM) is a very promising biomaterial for tissue engineering due to its easy accessibility, malleability and proven biological activity. In the present study, human dental pulp stem cells (hDPSCs) were combined in vitro with ECM scaffolds from porcine and human decellularised adipose tissues (pDAT, hDAT) processed as 3D solid foams, to investigate their effects on the osteogenic differentiation capacity and bone matrix production of hDPSCs, compared to single-protein-based 3D solid foams of collagen type I and conventional 2D tissue-culture-treated polystyrene plates. pDAT solid foams supported the osteogenic differentiation of hDPSCs to similar levels to collagen type I, as assessed by alkaline phosphatase and alizarin red stainings, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and osteocalcin/bone gamma-carboxyglutamate protein (BGLAP) immunostaining. Interestingly, hDAT solid foams showed a markedly lower capacity to sustain hDPSC osteogenic differentiation and matrix calcification and a higher capacity to support adipogenesis, as assessed by RT-qPCR and oil red O staining. White ATs from both human and porcine origins are relatively abundant and available sources of raw material to obtain high quality ECM-derived biomedical products. These biomaterials could have promising applications in tissue engineering and personalised clinical therapy for the healing and regeneration of lesions involving not only a loss of calcified bone but also its associated soft non-calcified tissues.

Immunoreaction to xenogenic tissue in cardiac surgery: alpha-Gal and beyond.

The ability of the immune system to distinguish self from non-self, reacting only to non-self, is essential for immune homeostasis. The mechanisms by which the immune system recognizes and responds to xenogenic tissue heart valves likely involve the initial activation of the innate immune system, with subsequent adaptive immune activation. The antigenicity of xenogeneic tissue constructs, primarily of porcine and bovine origin, has been extensively investigated and several antigenic epitopes have been identified. Some of them have been more exhaustively studied to avoid or at least reduce the immunoreaction triggered by their presence. Galactose-α-1,3-galactose (alpha-Gal) is a carbohydrate epitope present on the cells of all mammals (Fig. 1A). However, in ancestral Old World monkeys and apes, the gene for the enzyme alpha-1,3-galactosyltransferase, essential for the synthesis of alpha-Gal, resulted inactivated [1]. As a consequence, humans and recent Old World primates do not express the epitope. For xenotransplantation, i.e. transplantation of organs from pigs into humans, the mammals' anti-alpha-Gal antibodies represent a major immune barrier [1] (Fig. 1B).

Biomechanical Characterization of a New Acellular Dermal Matrix for Oral Soft Tissue Regeneration

Objectives Acellular dermal matrices (ADM) are a suitable alternative to autogenous soft tissue grafts (ASG). The aim of this study was to analyze the biomechanical properties and architectural features of ASG and ADM. Materials and methods ASG were harvested from the hard palate of fresh frozen body donors as connective tissue grafts and compared to ADM of porcine origin (NovoMatrix, NM; mucoderm, MD). Maximum load (ML, Newton [N]) and expansion (E, [mm]) were measured after rehydration in saline solution by tensile strength measurement. Light microscopy (LM) and scanning electron microscopy (SEM) were performed to analyze the architectural features of ASG and ADM in high resolution. Results ASG demonstrated a significantly decreased ML compared to NM and MD (p < 0.0001 and p = 0.019). NM showed a significantly increased ML compared to MD (p = 0.001). ASG demonstrated a non-significantly reduced E compared to NM (p = 0.13) and a significantly increased E compared to MD (p = 0.025). NM showed an increased E compared to MD (p < 0.0001). LM and SEM highlighted the surface characteristics and internal structures of ASG and ADM, such as the surface compact layer of MD and the densely packed, parallel running and ordered collagen fibers of NM and MD. Conclusions Significant differences concerning the biomechanical properties and architectural features of ASG, and ADM were found. Clinical relevance Information about the biomechanical properties and architectural features of ASG and ADM can contribute to a better understanding of the clinical performance and extend the application area.

Isolation, Characterization, and Molecular Detection of Porcine Sapelovirus.

Porcine sapelovirus (PSV) is an important emerging pathogen associated with a wide variety of diseases in swine, including acute diarrhoea, respiratory distress, skin lesions, severe neurological disorders, and reproductive failure. Although PSV is widespread, serological assays for field-based epidemiological studies are not yet available. Here, four PSV strains were recovered from diarrheic piglets, and electron microscopy revealed virus particles with a diameter of ~32 nm. Analysis of the entire genome sequence revealed that the genomes of PSV isolates ranged 7569–7572 nucleotides in length. Phylogenetic analysis showed that the isolated viruses were classified together with strains from China. Additionally, monoclonal antibodies for the recombinant PSV-VP1 protein were developed to specifically detect PSV infection in cells, and we demonstrated that isolated PSVs could only replicate in cells of porcine origin. Using recombinant PSV-VP1 protein as the coating antigen, we developed an indirect ELISA for the first time for the detection of PSV antibodies in serum. A total of 516 swine serum samples were tested, and PSV positive rate was 79.3%. The virus isolates, monoclonal antibodies and indirect ELISA developed would be useful for further understanding the pathophysiology of PSV, developing new diagnostic assays, and investigating the epidemiology of the PSV.

Development of a double-antibody sandwich ELISA targeting the receptor binding domain of TcdB toxin of ST11 type Clostridium difficile of porcine origin

Clostridium difficile is an important zoonotic intestinal pathogen, which is widely present in humans and a variety of animals. The ST11 type C. difficile is one of the most widespread and harmful subtypes in the world. As a large country in pig farming, China lacks efficient methods for detecting C. difficile of porcine origin, leaving hidden dangers for the prevention and control of C. difficile. The aim of this study was to develop a specific and sensitive double-antibody sandwich ELISA for the epidemiological investigation of ST11 type C. difficile of porcine origin. Firstly, a 97 kDa receptor binding domain (RBD) was expressed in a prokaryotic host and purified. A hybridoma cell line AE2D3 capable of stably secreting monoclonal antibody targeting the RBD was screened, and the antibody subtype was determined to be IgG2b (κ). Secondly, a double antibody sandwich ELISA method was developed, where the monoclonal antibody targeting the RBD was used as a detection antibody, and the rabbit polyclonal antibody was used as a capture antibody. The chessboard method was used to determine the matching concentration of the capture antibody and the detection antibody, the antigen coating conditions, the blocking conditions, the incubation conditions for detection antibody and samples to be tested, as well as the reaction conditions of HRP-conjugated and reaction conditions of TMB chromogenic solution. The negative cutoff OD450 was 0.152, and no cross-reaction with 13 strains of non-ST11 type C. difficile was found. The minimum detection concentration of RBD was 8.83 ng/mL. This specific and sensitive double-antibody sandwich ELISA provides a reliable serological detection method for epidemiological investigation of the ST11 type C. difficile in pig industry.

Indirect Immunoperoxidase Test (IPT) for Detection of Antibodies Against African Swine Fever Virus (ASFV) on African Green Monkey Cell Lines (Vero, MS).

The immunoperoxidase technique (IPT) is an immunocytochemistry technique on fixed infected cells to determine the antibody-antigen complex formation through the action of the peroxidase enzyme. In this procedure, African green monkey established cell lines, such as Vero or MS, are infected with African swine fever virus (ASFV)-adapted isolates to these cell cultures. The infected cells are fixed and then used as antigens to determine the presence of the specific antibodies against ASFV in samples of swine origin. IPT has many advantages for the serological diagnosis of ASF. Firstly, IPT has a higher sensitivity than ELISA or immunoblot assay (IB) for the detection of antibodies against ASFV. Secondly, it has greater versatility to analyze any type of clinical sample of porcine origin and to conduct research studies on the epidemiological situation, through the presence of antibody titers in exudates from organs and tissues. Thirdly, the preparation of IPT plates with cells infected with ASFV is simple, although laborious, that can be stored for a long time and allow the analysis of a large number of samples.

Leite com sabor fortificado com ferro heme: análise sensorial e de qualidade

Abstract The fortification of dairy beverages is a widely developed strategy using non-heme or heme iron. Heme iron has a higher bioavailability. The investigation aimed to elaborate pasteurized milk with fortified chocolate flavor with heme iron that has good sensory acceptability. The preparation of the flavored milk was carried out based on the regulations and heme iron, obtained from a commercial source of whole blood of porcine origin, was added before the pasteurization process to achieve its complete dilution. The concentration of iron and chocolate flavoring was established as variables in order to evaluate the optimal formulation based on Sensory Acceptability (SA). The experimental design was a 32 factorial design in which eight formulations were established, which were sensory acceptability evaluated by a total of 35 school-age children, aged between 8 and 11 years using a five-point facial hedonic scale. The results of the analysis of variance and optimization of the response showed that SA was 4.71 (on a scale of 1 to 5) for a fortification of 6.76 mg Fe kg-1 sample and a chocolate concentration of 2.0 g kg-1 sample. The physicochemical characterization indicated a higher percentage of carbohydrates, a higher concentration of iron (9.3 mg Fe kg-1 sample) and vitamin C (349.0 mg kg-1 sample) with respect to fresh milk. According to the physicochemical and microbiological results, the approximate life time of the beverage was 5 days, which is in accordance with Peruvian regulations. These results showed a method of fortification of flavored milk that allowed the use of heme iron, whose content could contribute to the daily requirement of this mineral in children aged between 8 and 11 years old (8 mg of iron per day).

EFECTO INHIBITORIO DE Lactobacillus plantarum AISLADOS DESDE LAS DISTINTAS ETAPAS DE LA CADENA PRODUCTIVA DE CARNE PORCINA FRENTE A Campylobacter coli

Lactobacillus plantarum has a great capacity to adapt to environmental and have antibacterial capacity against different pathogen. This is a potential alternative for the control of pathogen in food coming slaughter animal. The objective of this work was to determine the antibacterial effect of L. plantarum strains against C. coli strains. L. plantarum LP1, LP3, LP5 and LP7 of porcine origin and a reference strain of Campylobacter coli NCTC 11366 were used. C. coli and L. plantarum was reactivated. Lyophilized ELC and ELCn were obtained. C. coli was distributed on the surface of mCCDA by swab and the ELC and the ELCn were inoculated in wells previously made on the agar. After 48 h, the diameter of the zone of inhibition was measured. The inhibition halos produced were 23.2 mm. The ELCn showed less antagonistic activity. The LP5 ELC generated higher inhibition halos and ELCn showed no differences between the four strains studied. L. plantarum strains, isolated from the pig production chain, could potentially be applied to control C. coli. This data is added to the scarce scientific evidence of the inhibitory effect of L. plantarum before this zoonotic pathogen of importance in public health.

Genomic characterization of an African G4P[6] human rotavirus strain identified in a diarrheic child in Kenya: Evidence for porcine-to-human interspecies transmission and reassortment.

Human rotavirus strains having the unconventional G4P[6] genotype have been sporadically identified in diarrheic patients in different parts of the world. However, the whole genome of only one human G4P[6] strain from Africa (central Africa) has been sequenced and analyzed, and thus the exact origin and evolutionary pattern of African G4P[6] strains remain to be elucidated. In this study, we characterized the full genome of an African G4P[6] strain (RVA/Human-wt/KEN/KCH148/2019/G4P[6]) identified in a stool specimen from a diarrheic child in Kenya. Full genome analysis of strain KCH148 revealed a unique Wa-like genogroup constellation: G4-P[6]-I1-R1-C1-M1-A1-N1-T7-E1-H1. NSP3 genotype T7 is commonly found in porcine rotavirus strains. Furthermore, phylogenetic analysis showed that 10 of the 11 genes of strain KCH148 (VP7, VP4, VP6, VP1-VP3, NSP1, and NSP3-NSP5) appeared to be of porcine origin, the remaining NSP2 gene appearing to be of human origin. Therefore, strain KCH148 was found to have a porcine rotavirus backbone and thus is likely to be of porcine origin. Furthermore, strain KCH148 is assumed to have been derived through interspecies transmission and reassortment events involving porcine and human rotavirus strains. To our knowledge, this is the first report on full genome-based characterization of a human G4P[6] strain from east Africa. Our observations demonstrated the diversity of human G4P[6] strains in Africa, and provide important insights into the origin and evolutionary pattern of zoonotic G4P[6] strains on the African continent.

Linezolid-resistant Enterococcus gallinarum isolate of swine origin carrying cfr, optrA and poxtA genes.

To characterize a linezolid-resistant Enterococcus gallinarum isolate of porcine origin co-carrying cfr, optrA and poxtA genes. The genome was sequenced using the Illumina and Nanopore platforms. The presence of circular intermediates was examined by inverse PCR. Transferability of oxazolidinone resistance genes was investigated by transformation and conjugation. Two plasmids, the cfr- and optrA-carrying pEgFS4-1 (35 kb) and the poxtA-harbouring pEgFS4-2 (38 kb), were identified. pEgFS4-1 disclosed a distinctive mosaic structure with two cargo regions bounded by identical IS1216 elements interpolated into a backbone related to that of Enterococcus faecium vanA-containing pVEF2. The first cargo region included the cfr and optrA contexts, whereas the second one carried a Tn554 remnant and the lnu(A) gene. Both regions were able to excise in circular form as a unique translocable unit. pEgFS4-2 plasmid was 99% identical to a not fully described E. faecium pSBC1 plasmid. The poxtA environment, flanked by IS1216, was proved to be unstable. pEgFS4-2 also exhibited another cargo region containing the tet(M)-tet(L) genes arranged in tandem and its circular form was detected. Transformation and conjugation experiments failed to demonstrate the transferability of both plasmids to enterococcal recipients. Both plasmids persisted in the absence of selective pressure. To the best of our knowledge, this is the first description of a linezolid-resistant E. gallinarum isolate of swine origin carrying cfr, optrA and poxtA genes. The co-presence of three linezolid resistance determinants in an intrinsically vancomycin-resistant enterococcal species is cause of concern.

Screening novel CNS drug candidates for P-glycoprotein interactions using the cell line iP-gp: In vitro efflux ratios from iP-gp and MDCK-MDR1 monolayers compared to brain distribution data from mice

Drug efflux by P-glycoprotein (P-gp, ABCB1) is considered as a major obstacle for brain drug delivery for small molecules. P-gp-expressing cell monolayers are used for screening of new drug candidates during early states of drug development. It is, however, uncertain how well the in vitro studies can predict the in vivo P-gp mediated efflux at the blood–brain barrier (BBB). We previously developed a novel cell line of porcine origin, the iP-gp cell line, with high transepithelial resistance and functional expression of human P-gp. The aim of the present study was to evaluate the applicability of the cell line for screening of P-gp interactions of novel drug candidates. For this purpose, bidirectional fluxes of 14 drug candidates were measured in iP-gp cells and in MDCK-MDR1 cells, and compared with pharmacokinetic data obtained in male C57BL/6 mice. The iP-gp cells formed extremely tight monolayers (>15 000 Ω∙cm2) as compared to the MDCK- MDR1 cells (>250 Ω∙cm2) and displayed lower Papp,a-b values. The efflux ratios obtained with iP-gp and MDCK-MDR1 monolayers correlated with Kp,uu,brain values from the in vivo studies, where compounds with the lowest Kp,uu,brain generally displayed the highest efflux ratios. 12 of the tested compounds displayed a poor BBB penetration in mice as judged by Kp,uu less than 1. Of these compounds, nine compounds were categorized as P-gp substrates in the iP-gp screening, whereas analysis of data estimated in MDCK-MDR1 cells indicated four compounds as potential substrates. The results suggest that the iP-gp cell model may be a sensitive and useful screening tool for drug screening purposes to identify possible substrates of human P-glycoprotein.

Multi-Layer Technique (MLT) with Porcine Collagenated Cortical Bone Lamina for Bone Regeneration Procedures and Immediate Post-Extraction Implantation in the Esthetic Area: A Retrospective Case Series with a Mean Follow-Up of 5 Years.

Background: Augmentation of the edentulous atrophic anterior region is a challenging situation. The purpose of this article was to evaluate the effectiveness of a collagenated cortical bone lamina of porcine origin for horizontal ridge augmentation in patients with inadequate alveolar ridge width undergoing immediate post-extraction implantation in the anterior sites, and to report on implant survival rates/complications. Materials and methods: The cases were extracted electronically from a large database according to these specific inclusion criteria: patients with inadequate alveolar ridge width in the anterior maxilla or mandible, who underwent immediate post-extraction implant placement and simultaneous alveolar bone reconstruction using xenogeneic cortical bone lamina. An additional layer of palatal connective tissue graft was inserted between lamina and the vestibular mucosa, for improving soft tissue healing. A collagenated bone substitute was additionally placed in the gap between the lamina and implant surface in all patients. The main outcomes were implant survival and complications. Results: Forty-nine patients with 65 implants were included. Patients’ mean age at the time of implant surgery was 60.0 ± 13.6 years. The mean follow-up was 60.5 ± 26.6 months after implant placement. The implant survival was 100%. Four postoperative complications occurred in four patients. No specific factor was found to be associated with complication occurrence. Conclusion: The use of collagenated cortical bone lamina can be considered as a successful option for alveolar reconstruction in immediate post-extraction implant insertion procedures in anterior regions with inadequate alveolar ridge width.

Biological properties of swine vesicular disease virus strain 2348 Italy/2008

Swine vesicular disease (SVD) is a viral infectious disease, which, if acute, is manifested by the clinical pattern similar to a number of vesicular diseases including foot-and-mouth disease. In case of subclinical disease, there are no evident clinical signs, therefore the diagnosis is problematic, and there can be the risk of the disease introduction into the Russian Federation with the infected pigs. The key measure for the prevention of SVD introduction involves control diagnostic testing of all animals imported in the country that makes it necessary to keep updated the currently used methods and tools for the disease laboratory diagnosis. The paper demonstrates data on experimental infection of pigs with SVDV strain 2348 Italy/2008 that belongs to the most recent one of the four known phylogenetic groups. The virus was kindly provided by the World Reference Laboratory for Foot-and-Mouth Disease (Pirbright, Great Britain), and it was adapted to the monolayer continuous cell cultures of porcine origin (IB-RS-2 and PGSK-30). The pigs were intradermally infected with concentrated cultured virus at a dose of 109 TCID50. The infected animals demonstrated clinical signs typical for the acute disease. There was evidence that the virus was not transmitted to the intact animal in case husbandry conditions were met that allowed to avoid the infection transmission by the fecal-oral and contact mechanisms. As a result of the experiment, reference sera were collected at different time intervals post infection and their activity was determined using virus microneutralization test in cell culture and ELISA. Aphthae collected from the infected animals were deposited into the Strain collection of the Reference Laboratory for Foot-and-Mouth Disease, FGBI “ARRIAH”.

Full genome-based characterization of G4P[6] rotavirus strains from diarrheic patients in Thailand: Evidence for independent porcine-to-human interspecies transmission events.

The exact evolutionary patterns of human G4P[6] rotavirus strains remain to be elucidated. Such strains possess unique and strain-specific genotype constellations, raising the question of whether G4P[6] strains are primarily transmitted via independent interspecies transmission or human-to-human transmission after interspecies transmission. Two G4P[6] rotavirus strains were identified in fecal specimens from hospitalized patients with severe diarrhea in Thailand, namely, DU2014-259 (RVA/Human-wt/THA/DU2014-259/2014/G4P[6]) and PK2015-1-0001 (RVA/Human-wt/THA/PK2015-1-0001/2015/G4P[6]). Here, we analyzed the full genomes of the two human G4P[6] strains, which provided the opportunity to study and confirm their evolutionary origin. On whole genome analysis, both strains exhibited a unique Wa-like genotype constellation of G4-P[6]-I1-R1-C1-M1-A8-N1-T1-E1-H1. The NSP1 genotype A8 is commonly found in porcine rotavirus strains. Furthermore, on phylogenetic analysis, each of the 11 genes of strains DU2014-259 and PK2015-1-0001 appeared to be of porcine origin. On the other hand, the two study strains consistently formed distinct clusters for nine of the 11 gene segments (VP4, VP6, VP1-VP3, and NSP2-NSP5), strongly indicating the occurrence of independent porcine-to-human interspecies transmission events. Our observations provide important insights into the origin of zoonotic G4P[6] strains, and into the dynamic interaction between porcine and human rotavirus strains.

Treatment for rectourethral fistulas after radical prostatectomy with biological material interposition through a perineal access

IntroductionRectal injury is a rare complication after extraperitoneal laparoscopic radical prostatectomy. The development of rectourethral fistulas (URF) from rectal injuries is one of the most feared and of more complex resolution in urology. Material and methodsBetween 2013 and 2020 we have operated on a total of 5 patients with URF after extraperitoneal endoscopic radical prostatectomy through a perineal access using the interposition of biological material. All fistulas had a diameter of less than 6 mm at endoscopy and were less than 6 cm apart from the anal margin. ResultsThe mean age of the patients was 64 years old. All patients had a previous bowel and urinary diversion for at least 3 months. Under general anesthesia and with the patient in a forced lithotomy position, fistulorraphy and interposition of biological material of porcine origin (lyophilized porcine dermis [Permacol®]) were performed through a perineal access. Mean operative time was 174 min (140–210). Most patients were discharged on the third postoperative day. The bladder catheter was left in place for a mean of 40 days (30–60). Prior to its removal, cystography and a Gastrografin® barium enema were performed, showing resolution of the fistula in all cases. ConclusionsThe interposition of biological material from porcine dermis through perineal approach is a safe alternative with good results in patients submitted to urethrorectal fistulorraphy after radical prostatectomy.