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1.
J Cardiothorac Surg ; 19(1): 43, 2024 Feb 03.
Article in English | MEDLINE | ID: mdl-38310241

ABSTRACT

BACKGROUND: Tumor-like lesions of the trachea are rare and challenging in diagnosis and management. Inflammatory myofibroblastoma, also known as Inflammatory pseudo tumors (IPTs), as well as Rosai Dorfman Disease (RDD) are inflammatory lesions that may involve the central airways with variable non-specific clinical features mimicking tumors. CASE PRESENTATION: In this study 2 cases with tumor-like lesions are presented. One case with an inflammatory pseudotumor and the other one with Rosai-Dorfman disease affecting the upper trachea. Both cases were successfully managed with tracheal resection anastomosis. CONCLUSION: Tracheal Inflammatory myofibroblastoma, and Rosai-Dorfman diseases are rare tumor like lesions that present with upper airway obstruction. Despite being benign, these lesions may have features suggestive of malignancy, requiring prompt management. Complete surgical excision by segmental resection and primary anastomosis (if feasible) is the treatment of choice with an optimum outcome.


Subject(s)
Histiocytosis, Sinus , Neoplasms, Muscle Tissue , Neoplasms , Humans , Histiocytosis, Sinus/diagnosis , Trachea/surgery , Trachea/pathology , Anastomosis, Surgical
2.
Reprod Domest Anim ; 57(4): 402-417, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34971452

ABSTRACT

The present study aimed to use a comprehensive approach for evaluating 12 factors related to embryo quality (shape, size and transparency), donors (age, ET type, number of recovered embryos and day of uterine flushing), recipients (age), males (age and individual variations) and environment (season and year) which could affect the outcome of ET in terms of pregnancy (PR) and pregnancy losses in dromedary camels. During three breeding seasons, 116 donor females were mated repeatedly at 12- to 14-day intervals by fertile male camels (n = 33) without stimulation of the ovaries (WSPO). Superovulation (SPO ET) regimen was applied for each donor female twice or thrice per season. In the occasions of applying superovulation regimen, donor females having an ovulatory follicle were mated instead of GnRH administration and superovulation regimen was applied 4 days post-mating (MIX ET). The uteri of all donor females were flushed at Day 8 or 9 post-mating, and a total of 2,095 embryos were recovered and transferred individually to 924 recipient females. Pregnancy diagnoses were conducted at Day 10 after ET (Days 18-19 of gestation) by using progesterone assay and by transrectal ultrasonography (TRU) at Days 30 and 60 of gestation. By using logistic regression analysis, transparency of embryos and age of recipient females had significant effects on PR at Days 18-19 (p < .01), 30 (p < .01) and 60 (p < .01; p < .05, respectively) of gestation. The shape of embryos had significant effects on the PR at Days 30 (p < .05) and 60 (p < .01) of gestation. Type of ET and the breeding season (year) had significant (p < .05) effects on the PR at Day 30, while day of flushing had the same effect on PR at Day 60. Regarding the pregnancy losses, transparency and shape of the embryo, type of ET, breeding season had significant (p < .05) effect on the late embryonic mortalities (LEM) and shape and season of year had significant (p < .01 and p < .05, respectively) effect on LEM/early foetal mortalities (EFM). Regarding male individual factor, there was a tendency for a significant (p = .055) effect of male camels on the PR at Days 18-19 and rate of LEM. In conclusion, transferring a spherical, transparent or a large-sized embryo (>750 µm) into recipient females ageing between 8 and 11 y could greatly improve the PR from Days 18 to 60 of gestation. Also, embryo recovered from donor females with Mix ET type or embryos sired by certain male camel or at Day 8 post-mating of the donor could improve the 2-month PR. In addition, transferring a transparent or spherical-shaped embryo or embryos recovered from donor females with SPO or Mix ET could reduce the pregnancy losses during the first 2 months of pregnancy.


Subject(s)
Abortion, Veterinary , Camelus , Animals , Camelus/physiology , Embryo Transfer/veterinary , Female , Male , Pregnancy , Retrospective Studies , Superovulation
3.
Theriogenology ; 143: 44-49, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31835099

ABSTRACT

The aims of the present study were to evaluate the effect of cooling of the dromedary camel embryos on the pregnancy and pregnancy loss rates, and to investigate the factors which might affect the outcomes of the transfer of cooled embryos. After the donors (n = 56) had been super-ovulated and mated, they were flushed at Day 8 or 9 post-mating. Of 487 collected embryos, 110 were refrigerated at 4°C for up to 5 days in holding medium (HM), flushing medium supplemented with 10% fetal calf serum (FM + FCS) or TCM199 supplemented with 50% FCS and HEPES (TCM + FCS + HEPES). Both fresh (n = 377) and cooled embryos were transferred individually into synchronized recipients. Pregnancy diagnoses were carried out at Days 18-19, 30 and 60 post-mating of the donors. Transferring of fresh embryos into the recipients resulted in significantly higher pregnancy rates at Days 18-19 (53.1% vs. 38.2%, P < 0.01), Day 30 (46.4% vs. 31.8%, P < 0.01) and Day 60 (42.4% vs. 26.4%, P < 0.005) compared with those of cooled embryos, respectively. Pregnancy rates after transferring cooled embryos progressively decreased with the prolongation of the storage period. A significant difference in the pregnancy rate (56% vs. 13%, respectively, P < 0.05) was recorded only at Days 18-19 between cooled embryos held for one day and those held for 5 days. The pregnancy rates at Days 18-19, Day 30 and Day 60 were non-significantly higher when TCM + HEPES and FCS medium used for cooling of embryos compared to those of FM + FCS or HM medium. Cooling of spherical embryos resulted in significantly higher pregnancy rates at Days 30 (45.6% vs. 17.0%, respectively, P < 0.005) and 60 (42.1 vs. 9.4%, respectively, P < 0.005) and a significantly lower pregnancy loss rate (11.1% vs. 66.6%, respectively, P < 0.005) compared to those resulting from cooling of folded embryos. Neither the size of embryo nor the day of flushing had a significant effect on the pregnancy and pregnancy loss rates after the transfer of cooled embryos. In conclusion, pregnancy could be obtained after the transfer of camel embryos refrigerated for up to 5 days. In addition, higher pregnancy rates could be obtained if only spherical embryos are selected for cooling.


Subject(s)
Abortion, Veterinary , Camelus/embryology , Embryo, Mammalian , Tissue Preservation , Animals , Embryo Culture Techniques , Embryo Transfer/veterinary , Female , Fertilization in Vitro , Pregnancy
4.
J Orthop Surg (Hong Kong) ; 27(3): 2309499019861031, 2019.
Article in English | MEDLINE | ID: mdl-31315494

ABSTRACT

PURPOSE: The aim of this study is to report the clinical and radiological outcomes following surgical treatment of chondroblastoma by means of an extended intralesional curettage using high-speed burr, with phenol as a local adjuvant which is followed by the implantation of synthetic bone graft, aiming to lower the recurrence rate of this tumor. PATIENTS AND METHODS: This retrospective study included 20 patients with chondroblastoma lesions during the period between 2000 and 2015. RESULTS: Fifteen males and five females were followed up for a mean of 63.35 (26-144) months with average age at the time of presentation was 20.8 (range: 12-32) years. Nineteen patients (95%) were complaining of pain at the time of presentation, and the lesion was discovered accidently in one patient. The mean operative time was 138.5 min (75-250). At the most recent follow-up, all patients had regained full physical function without pain at the operation site. CONCLUSION: The aggressive treatment of chondroblastoma by an extended intralesional curettage using high-speed burr with phenol as a local adjuvant seems effective in lowering the incidence of local recurrence and secondary more aggressive surgeries. Implantation of the bone defects that result from curettage with the synthetic bone substitutes is a good alternative due to rapid restoration of the mechanical strength with good remodeling.


Subject(s)
Bone Neoplasms/surgery , Bone Transplantation/methods , Chondroblastoma/surgery , Curettage/methods , Phenol/pharmacology , Adolescent , Adult , Anti-Infective Agents, Local/pharmacology , Child , Female , Humans , Male , Radiography , Retrospective Studies , Treatment Outcome , Young Adult
5.
J Orthop Surg (Hong Kong) ; 27(1): 2309499019832970, 2019.
Article in English | MEDLINE | ID: mdl-30879390

ABSTRACT

Due to accurate preoperative imaging techniques, early diagnosis and effective chemotherapy, many tumors arising in the metaphyseo-diaphyseal regions of long bones can be segmentally resected with joint preservation. The intercalary resection of malignant bone tumor results in a bone defect which can represent a challenging reconstructive problem. The most commonly used surgical reconstructive options for these defects include biologic reconstructions such as allografts, vascularized fibular grafts, autogenous extracorporeally devitalized tumor bearing bone graft, combination of allografts or devitalized autografts with vascularized fibular grafts, segmental bone transport, or induced membrane technique. Nonbiologic reconstructions, on the other hand, use intercalary endoprostheses. Every patient should be carefully evaluated and the reconstructive option should be individually selected. The aim of this article is to discuss the surgical options of reconstruction of bone defects after intercalary resection of malignant bone tumors with reviewing of their indications, advantages, disadvantages and complications.


Subject(s)
Bone Neoplasms/surgery , Bone Transplantation/methods , Plastic Surgery Procedures/methods , Autografts , Bone Neoplasms/pathology , Fibula/transplantation , Humans , Prostheses and Implants , Transplantation, Homologous
6.
J Orthop Surg Res ; 13(1): 270, 2018 Oct 29.
Article in English | MEDLINE | ID: mdl-30373674

ABSTRACT

BACKGROUND: Benign bone tumors and tumor-like conditions are commonly located in the proximal femur. The main indications for surgical treatment are lesions with impending or actual pathological fractures, or with aggressive or recurrent lesions. However, patients complaining of persistent pain, limping, or abnormal gait patterns are also considered for surgical treatment. In this study, we describe the outcomes of the surgical treatment of benign lytic lesions of the proximal femur by curettage followed by implantation of synthetic bone graft. METHODS: This retrospective study included 27 patients (22 females and 5 males) with benign lytic lesions of the proximal femur. The average age was 25.5 years (6-65 years), and the mean follow-up period was 54.5 months (9-145 months). The histopathological diagnoses were fibrous dysplasia (8 patients), simple bone cyst (8 patients), chondroblastoma (7 patients), giant cell tumor (3 patients), and eosinophilic granuloma (1 patient). These lesions were managed with curettage followed by implantation of the bone defects with alpha tricalcium phosphate in 14 patients, beta tricalcium phosphate granules in 11 patients, hydroxyapatite granules in 1 patient, and combined beta tricalcium phosphate and hydroxyapatite granules in 1 patient. Internal fixation was performed in three patients. RESULTS: The mean operative time was 143 min (80-245 min). Patients had regained normal unrestricted activity without pain at the operation site. Patients treated with beta tricalcium phosphate achieved radiographic consolidation of the bone defects within 1 year after the surgery, and those treated with alpha tricalcium phosphate or hydroxyapatite experienced no progression nor recurrence of the lesions. Local tumor recurrence was observed in one patient with giant cell tumor 5 years after the surgery. Post-operative pathological fracture was occurred in one patient with a simple bone cyst of the subtrochanteric region 1 month after surgery. No post-operative infection was observed. CONCLUSION: We concluded that the treatment of benign lytic lesions of the proximal femur, either primary or recurrent, using synthetic bone graft is a safe and satisfactory method and the addition of internal fixation should be carefully planned.


Subject(s)
Bone Diseases/surgery , Bone Transplantation , Femur/surgery , Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Retrospective Studies , Young Adult
7.
Parasit Vectors ; 11(1): 390, 2018 Jul 04.
Article in English | MEDLINE | ID: mdl-29973271

ABSTRACT

BACKGROUND: To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options. METHODS: To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. Systemic production of Th1, Th2, inflammatory and regulatory cytokines was determined in different immune compartments at mRNA and (after stimulation with a recombinant merozoite-protein) at protein level by PCR and multiplex fluorescent immunoassay, respectively. RESULTS: Infection generated significantly increased serum IgA and IgG levels against C. suis sporozoites and merozoites, irrespective of infection mode, with IgG against merozoites showing the strongest increase. No clinical signs and only occasional excretion were observed. The systemic cytokine response to C. suis was only weak. Nonetheless, in white blood cells, IL-4, IL-6 and IL-10 mRNA-levels significantly increased after infection, whereas IFN-É£, IL-2 and TGF-ß expression tended to decrease. In mesenteric lymph nodes (MLN), IL-10 and TNF-α levels were elevated while splenic cytokine expression was unaltered upon infection. Stimulated MLN-derived lymphocytes from infected pigs produced slightly more IL-12 and less IFN-α than controls. CONCLUSIONS: An infection and a subsequent systemic immune response can be induced in immune-competent animals by all evaluated infection models and growers can be used as models to mimic sow immunizations. The immune response to C. suis, although mild and with considerable variation in cytokine expression, was characterized by a Th2-associated and regulatory cytokine profile and antibody production. However, none of the parameters clearly stood out as a potential marker associated with protection. Antibody titers were significantly positively related with oocyst excretion and might thus serve as correlates for parasite replication or severity of infection.


Subject(s)
Antibodies, Protozoan/immunology , Coccidiosis/immunology , Cytokines/immunology , Sarcocystidae/immunology , Swine Diseases/immunology , Age Factors , Animals , Antibodies, Protozoan/biosynthesis , Antibodies, Protozoan/blood , Cytokines/biosynthesis , Cytokines/blood , Cytokines/genetics , Feces/parasitology , Female , Immunocompetence , Immunoglobulin G/blood , Merozoites/immunology , Oocysts/immunology , Parasite Egg Count , Sporozoites/immunology , Swine , Swine Diseases/parasitology , Th2 Cells/immunology
8.
Eur Arch Otorhinolaryngol ; 275(9): 2245-2252, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29982939

ABSTRACT

BACKGROUND: Spontaneous CSF leak is a challenging condition, with frequent recurrences following attempted surgical closure. The selection of graft materials depends on the experience of the operating surgeon. Leukocyte- and platelet-rich fibrin (L-PRF) is a second-generation platelet concentrate that has currently invaded every surgical specialty. To our knowledge, no previous reports about the use of L-PRF in repair of CSF leak were published. The study was conducted to assess the potential role of L-PRF in spontaneous CSF leaks repair. METHODS: This prospective controlled study was conducted on 40 patients who were randomly divided into 2 groups: In group A, L-PRF was used in addition to another layer, while patients in group B underwent multilayer repair using traditional graft materials. RESULTS: Preoperative HRCT scan showed bony dehiscence (n = 30) and opacity in a nearby sinus (n = 9). MRI findings varied between empty sella in 28 patients, and meningocele in 19 patients. Mean preoperatively measured CSF pressure was 26.6 cm H2O. Cribriform plate was identified as the commonest site of leaks. Multiple skull defects were detected in 2 cases. Failure of repair was found in 3 patients (15%) of group B and 1 patient (5%) of group A, this difference was statistically insignificant (P value = 0.28). Significance correlation between recurrence of CSF leak and elevated intracranial pressure was observed (P value; 0.04). CONCLUSION: L-PRF was found to be a good adjunct material in endoscopic spontaneous CSF leaks repair. It helps in decreasing the number of layers needed for defect closure.


Subject(s)
Cerebrospinal Fluid Rhinorrhea/therapy , Natural Orifice Endoscopic Surgery , Platelet-Rich Fibrin , Adult , Cerebrospinal Fluid Rhinorrhea/diagnostic imaging , Cerebrospinal Fluid Rhinorrhea/etiology , Female , Humans , Intracranial Hypertension/complications , Magnetic Resonance Imaging , Male , Middle Aged , Prospective Studies , Recurrence , Tomography, X-Ray Computed , Treatment Outcome , Young Adult
9.
Vet Res ; 48(1): 19, 2017 04 05.
Article in English | MEDLINE | ID: mdl-28381233

ABSTRACT

Fredericella sultana is an invertebrate host of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease in salmonids. The bryozoan produces seed-like statoblasts to facilitate its persistence during unfavourable conditions. Statoblasts from infected bryozoans can harbor T. bryosalmonae and give rise to infected bryozoan colonies when conditions improve. We aimed in the present study to evaluate the integrity and viability of T. bryosalmonae-infected statoblasts after a range of harsh treatment conditions. We tested if statoblasts could survive ingestion by either brown trout or common carp. After ingestion, the fish faeces was collected at different time points. We also tested physical stressors: statoblasts collected from infected colonies were desiccated at room temperature, or frozen with and without Bryozoan Medium C (BMC). After treatments, statoblasts were assessed for physical integrity before being incubated on BMC to allow them to hatch. After 4 weeks, hatched and unhatched statoblasts were tested by PCR for the presence of the parasite. We found that statoblasts ingested by brown trout and those frozen in BMC were completely broken. In contrast, statoblasts ingested by common carp and those subjected to dry freezing were able to survive and hatch. T. bryosalmonae was detected by PCR in both hatched and unhatched infected statoblasts, but neither from broken nor uninfected statoblasts. Our results confirmed for the first time the ability of infected statoblasts to survive passage through a fish, and freezing. These findings suggest potential pathways for both persistence and spread of T. bryosalmonae-infected statoblasts in natural aquatic systems.


Subject(s)
Bryozoa/parasitology , Fish Diseases/parasitology , Myxozoa/physiology , Animals , Bryozoa/cytology , Carps/parasitology , Feces/parasitology , Trout/parasitology
10.
Int Orthop ; 41(7): 1481-1487, 2017 07.
Article in English | MEDLINE | ID: mdl-28343291

ABSTRACT

PURPOSE: In 1999, we developed a technique using frozen autografts-tumour-containing bone treated with liquid nitrogen-for the reconstruction of malignant bone tumours. The purpose of this study was to evaluate the functional and oncological outcomes of frozen autografts for intercalary reconstruction of malignant bones and soft tissue tumours. METHODS: This retrospective study was designed to assess 34 patients of mean age 35 (range, 6-79) years. The mean follow-up period was 62 (24-214) months. The median length of the frozen autografts was 138.4 ± 60.39 (50-290) mm. RESULTS: Postsurgically, 20 patients remained disease-free, seven patients survived with no evidence of disease, five patients were alive with disease, and two patients died of disease. The five- and ten-year survival rates of the frozen autografts were 91.2% and the mean International Society of Limb Salvage score was 90%. Complete bony union was achieved in 97% of the patients. There were five cases of nonunion, six cases of fracture, two cases of deep infection and four cases of local recurrence. CONCLUSION: Utilizing intercalary frozen autografts for patients with a nonosteolytic primary or secondary bone tumour without involvement of the subchondral bone is a good alternative treatment, because it is a straightforward biological technique and can provide excellent limb function.


Subject(s)
Bone Neoplasms/surgery , Bone Transplantation/methods , Plastic Surgery Procedures/methods , Soft Tissue Neoplasms/surgery , Transplantation, Autologous/methods , Adolescent , Adult , Aged , Autografts , Bone Neoplasms/mortality , Bone Transplantation/adverse effects , Bone and Bones/pathology , Child , Female , Follow-Up Studies , Freezing , Humans , Limb Salvage/methods , Male , Middle Aged , Plastic Surgery Procedures/adverse effects , Retrospective Studies , Soft Tissue Neoplasms/mortality , Survival Analysis , Survival Rate , Transplantation, Autologous/adverse effects , Young Adult
11.
Parasit Vectors ; 10(1): 68, 2017 02 07.
Article in English | MEDLINE | ID: mdl-28173829

ABSTRACT

BACKGROUND: The genome of the apicomplexan parasite Cystoisospora suis (syn. Isospora suis) has recently been sequenced and annotated, opening the possibility for the identification of novel therapeutic targets against cystoisosporosis. It was previously proposed that a 42 kDa uncharacterized merozoite protein, encoded by gene CSUI_005805, might be a relevant vaccine candidate due to its high immunogenic score, high expression level and species-specificity as determined in silico. METHODS: The 1170 bp coding sequence of the CSUI_005805 gene was PCR amplified and cloned into the bacterial expression vector pQE-31. The specificity of the expressed recombinant protein was evaluated in an immunoblot, and relative levels of expression in different developmental stages and subcellular localization were determined by quantitative real-time PCR and indirect immunofluorescence assay, respectively. RESULTS: The CSUI_005805 gene encoded for a 389 amino acid protein containing a histidine-rich region. Quantitative RT-PCR showed that CSUI_005805 was differentially expressed during the early development of C. suis in vitro, with higher transcript levels in merozoites compared to sporozoites. The recombinant protein was specifically recognized by sera from chicken immunized with recombinant CSUI_005805 protein and sera from piglets experimentally infected with C. suis, all of which suggested that despite prokaryotic expression, the recombinant CSUI_005805 protein maintained antigenic determinants and could elicit an immune response in the host. Immunofluorescence labelling and confocal microscopy revealed localization primarily at the surface of the parasite. CONCLUSIONS: The results suggest that CSUI_005805 is highly expressed in merozoites and might thus be critical for their survival and establishment inside host cells. Owing to its specificity, localization and expression pattern, CSUI_005805 could be exploited as an attractive candidate for alternative control strategies against C. suis such as vaccines.


Subject(s)
Cloning, Molecular , Eimeriidae/genetics , Eimeriidae/metabolism , Gene Expression Regulation/physiology , Merozoites/metabolism , Protozoan Proteins/metabolism , Amino Acid Sequence , Animals , Base Sequence , DNA, Complementary/genetics , DNA, Protozoan/genetics , Polymerase Chain Reaction , Protozoan Proteins/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Protozoan/genetics , Sporozoites/metabolism
12.
Front Vet Sci ; 2: 68, 2015.
Article in English | MEDLINE | ID: mdl-26664994

ABSTRACT

Cystoisospora suis is a coccidian species that typically affects suckling piglets. Infections occur by oral uptake of oocysts and are characterized by non-hemorrhagic transient diarrhea, resulting in poor weight gain. Apparently, primary immune responses to C. suis cannot readily be mounted by neonates, which contributes to the establishment and rapid development of the parasite, while in older pigs age-resistance prevents disease development. However, the presence of extraintestinal stages, although not unequivocally demonstrated, is suspected to enable parasite persistence together with the induction and maintenance of immune response in older pigs, which in turn may facilitate the transfer of C. suis-specific factors from sow to offspring. It is assumed that neonates are particularly prone to clinical disease because infections with C. suis interfere with the establishment of the gut microbiome. Clostridia have been especially inferred to profit from the altered intestinal environment during parasite infection. New tools, particularly in the area of genomics, might illustrate the interactions between C. suis and its host and pave the way for the development of new control methods not only for porcine cystoisosporosis but also for other mammalian Cystoisospora infections. The first reference genome for C. suis is under way and will be a fertile ground to discover new drugs and vaccines. At the same time, the establishment and refinement of an in vivo model and an in vitro culture system, supporting the complete life cycle of C. suis, will underpin the functional characterization of the parasite and shed light on its biology and control.

13.
Parasitol Res ; 114(3): 929-39, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25563603

ABSTRACT

Tetracapsuloides bryosalmonae Canning et al., 1999 (Myxozoa) is the causative agent of proliferative kidney disease in various species of salmonids in Europe and North America. We have shown previously that the development and distribution of the European strain of T. bryosalmonae differs in the kidney of brown trout (Salmo trutta) Linnaeus, 1758 and rainbow trout (Oncorhynchus mykiss) Walbaum, 1792, and that intra-luminal sporogonic stages were found in brown trout but not in rainbow trout. We have now compared transcriptomes from kidneys of brown trout and rainbow trout infected with T. bryosalmonae using suppressive subtractive hybridization (SSH). The differentially expressed transcripts produced by SSH were cloned, transformed, and tested by colony PCR. Differential expression screening of PCR products was validated using dot blot, and positive clones having different signal intensities were sequenced. Differential screening and a subsequent NCBI-BLAST analysis of expressed sequence tags revealed nine clones expressed differently between both fish species. These differentially expressed genes were validated by quantitative real-time PCR of kidney samples from both fish species at different time points of infection. Expression of anti-inflammatory (TSC22 domain family protein 3) and cell proliferation (Prothymin alpha) genes were upregulated significantly in brown trout but downregulated in rainbow trout. The expression of humoral immune response (immunoglobulin mu) and endocytic pathway (Ras-related protein Rab-11b) genes were significantly upregulated in rainbow trout but downregulated in brown trout. This study suggests that differential expression of host anti-inflammatory, humoral immune and endocytic pathway responses, cell proliferation, and cell growth processes do not inhibit the development of intra-luminal sporogonic stages of the European strain of T. bryosalmonae in brown trout but may suppress it in rainbow trout.


Subject(s)
Fish Diseases/parasitology , Gene Expression Regulation/physiology , Myxozoa/physiology , Trout/metabolism , Animals , Base Sequence , Europe/epidemiology , Fish Diseases/epidemiology , Fish Diseases/metabolism , Kidney/metabolism , Kidney/parasitology , Kidney Diseases/epidemiology , Kidney Diseases/parasitology , Kidney Diseases/veterinary , North America/epidemiology , Polymerase Chain Reaction , Real-Time Polymerase Chain Reaction , Species Specificity
14.
Vet Res ; 45: 101, 2014 Oct 04.
Article in English | MEDLINE | ID: mdl-25297457

ABSTRACT

Tetracapsuloides bryosalmonae (Myxozoa) is the causative agent of proliferative kidney disease in various species of salmonids in Europe and North America. In Europe, spores of T. bryosalmonae develop in the kidney of infected brown trout Salmo trutta and are released via urine to infect the freshwater bryozoan Fredericella sultana. The transcriptomes of kidneys of infected and non-infected brown trout were compared by suppressive subtractive hybridization. Differential screening and a subsequent NCBI BLAST analysis of expressed sequence tags revealed 21 transcripts with functions that included cell stress and cell growth, ribonucleoprotein, signal transduction, ion transporter, immune response, hemoglobin and calcium metabolisms. Quantitative real time PCR was used to verify the presence of these selected transcripts in brown trout kidney at sporogonic stages of T. bryosalmonae development. Expression of cold-inducible RNA-binding protein, cyclin-dependent kinase inhibitor 2A, prothymosin alpha, transforming protein RhoA, immunoglobulin light chain and major histocompatibility complex class I were up-regulated significantly in infected brown trout. Expression of both the hemoglobin subunit beta and stanniocalcin precursor were down-regulated significantly in infected brown trout. This study suggests that cell stress and cell growth processes, signal transduction activities, erythropoiesis and calcium homeostasis of the host are modulated during sporogonic stages of parasite development, which may support the sporogenesis of T. bryosalmonae in the kidney of brown trout.


Subject(s)
Fish Diseases/immunology , Fish Proteins/genetics , Host-Pathogen Interactions/physiology , Myxozoa/physiology , Parasitic Diseases, Animal/immunology , Transcriptome , Trout , Animals , Expressed Sequence Tags , Fish Proteins/metabolism , Kidney/metabolism , Kidney/parasitology , Molecular Sequence Data , Real-Time Polymerase Chain Reaction/veterinary , Sequence Analysis, DNA/veterinary , Subtractive Hybridization Techniques/veterinary
15.
Dis Aquat Organ ; 111(1): 41-9, 2014 Aug 21.
Article in English | MEDLINE | ID: mdl-25144116

ABSTRACT

Proliferative kidney disease (PKD) is a widespread disease of farmed and wild salmonid populations in Europe and North America, caused by the myxozoan parasite Tetracapsuloides bryosalmonae. Limited studies have been performed on the epidemiological role in spread of the disease played by fish that survive infection with T. bryosalmonae. The aim of the present study was to evaluate the persistence of T. bryosalmonae developmental stages in chronically infected brown trout Salmo trutta up to 2 yr after initial exposure to laboratory-infected colonies of the parasite's alternate host, the bryozoan Fredericella sultana. Kidney, liver, spleen, intestine, brain, gills and blood were sampled 24, 52, 78 and 104 wk post-exposure (wpe) and tested for T. bryosalmonae by PCR and immunohistochemistry (IHC). Cohabitation trials with specific pathogen free (SPF) F. sultana colonies were conducted to test the viability of T. bryosalmonae. PCR detected T. bryosalmonae DNA in all tissue samples collected at the 4 time points. Developmental stages of T. bryosalmonae were demonstrated by IHC in most samples at the 4 time points. Cohabitation of SPF F. sultana with chronically infected brown trout resulted in successful transmission of T. bryosalmonae to the bryozoan. This study verified the persistence of T. bryosalmonae in chronically infected brown trout and their ability to infect the bryozoan F. sultana up to 104 wpe.


Subject(s)
Fish Diseases/parasitology , Myxozoa/physiology , Parasitic Diseases, Animal/parasitology , Trout , Animals , Chronic Disease , Immunohistochemistry/veterinary , Polymerase Chain Reaction/veterinary
16.
Parasitology ; 141(4): 482-90, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24229733

ABSTRACT

The freshwater bryozoan, Fredericella sultana, is the main primary host of the myxozoan endoparasite, Tetracapsuloides bryosalmonae which causes proliferative kidney disease (PKD) of salmonid fish. Because spores that develop in bryozoan colonies are infectious to fish, bryozoans represent the ultimate source of PKD. Bryozoans produce numerous seed-like dormant stages called statoblasts that enable persistence during unfavourable conditions and achieve long-distance dispersal. The possibility that T. bryosalmonae may undergo vertical transmission via infection of statoblasts has been the subject of much speculation since this is observed in close relatives. This study provides the first evidence that such vertical transmission of T. bryosalmonae is extensive by examining the proportions of infected statoblasts in populations of F. sultana on two different rivers systems and confirms its effectiveness by demonstrating transmission from material derived from infected statoblasts to fish hosts. Vertical transmission in statoblasts is likely to play an important role in the infection dynamics of both bryozoan and fish hosts and may substantially contribute to the widespread distribution of PKD.


Subject(s)
Bryozoa/parasitology , Fish Diseases/transmission , Infectious Disease Transmission, Vertical/veterinary , Myxozoa/isolation & purification , Parasitic Diseases, Animal/transmission , Salmonidae/parasitology , Animals , Female , Fish Diseases/parasitology , Fresh Water/parasitology , Myxozoa/genetics , Myxozoa/physiology , Parasitic Diseases, Animal/parasitology , Plant Roots/parasitology , Specific Pathogen-Free Organisms , Trees
17.
Dis Aquat Organ ; 107(1): 9-18, 2013 Nov 25.
Article in English | MEDLINE | ID: mdl-24270019

ABSTRACT

Tetracapsuloides bryosalmonae (Myxozoa) is the causative agent of proliferative kidney disease in salmonids. We assessed differences in intensity of T. bryosalmonae infection between brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss from the clinical phase of infection onwards. Specific pathogen-free fish were exposed to T. bryosalmonae spores under controlled laboratory conditions and sampled at 6, 8, 10, 12, 14, and 17 wk post exposure (wpe), and the transmission of T. bryosalmonae from infected fish to the bryozoan Fredericella sultana was observed. Parasite load was determined in fish kidneys by quantitative real-time PCR (qRT-PCR), and parasite stages were detected in kidney, liver, and spleen tissues at different time points by immunohistochemistry. T. bryosalmonae was successfully transmitted from infected brown trout to F. sultana colonies but not from infected rainbow trout. Body length and weight of infected brown trout did not differ significantly from control brown trout during all time points, while length and weight of infected rainbow trout differed significantly compared to controls from 10 to 17 wpe. qRT-PCR revealed that parasite load was significantly higher in kidneys of brown trout compared with rainbow trout. Immunohistochemistry showed high numbers of intra-luminal stages (sporogonic stages) in kidneys of brown trout with low numbers of pre-sporogonic stages. Sporogonic stages were not seen in kidneys of rainbow trout; only high numbers of pre-sporogonic stages were detected. Numbers of pre-sporogonic stages were low in the spleen and liver of brown trout but high in rainbow trout. These data confirmed that there are differences in the development and infection progress of T. bryosalmonae between brown trout and rainbow trout.


Subject(s)
Fish Diseases/parasitology , Myxozoa , Oncorhynchus mykiss , Parasitic Diseases, Animal/parasitology , Animals , Immunohistochemistry , Kidney Diseases/parasitology , Kidney Diseases/veterinary , Species Specificity , Specific Pathogen-Free Organisms , Time Factors
18.
Eur Arch Otorhinolaryngol ; 267(10): 1569-72, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20383515

ABSTRACT

The objective of the study was conducted to evaluate the effectiveness of nasal endoscopy for both diagnosis and localized excision of intranasal contact areas that cause headache and/or facial pain as well as to evaluate the use of lidocaine test for diagnosis of such cases and predicting the result of surgery. This study included 120 patients aged between 18 and 45 years, with an average period of headache and/or facial pain of 2.5 years. Patients were classified into two groups according to lidocaine test. Group A that was lidocaine positive and group B which was negative. Excision of contact points was done, under endoscopic guidance, from the septum as well as the lateral nasal wall. 98.75% of patients in group A got benefit from surgery as most cases were cured from headache and facial pain. In group B, 40% got benefit with most patients had unchanged symptoms. In conclusion, endoscopic surgery gives a more precise complete excision of limited areas without time wasting or morbidity. Lidocaine test can be used as a test to aid in diagnosis and to predict the result of surgery where there were high cure rate within the lidocaine-positive group.


Subject(s)
Anesthetics, Local , Endoscopy , Facial Pain/surgery , Headache/surgery , Lidocaine , Nasal Mucosa/pathology , Adolescent , Adult , Facial Pain/etiology , Facial Pain/pathology , Female , Headache/etiology , Headache/pathology , Humans , Male , Middle Aged , Nasal Mucosa/surgery , Nasal Septum/pathology , Nasal Septum/surgery , Predictive Value of Tests , Treatment Outcome , Young Adult
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