Immunization with thiol-binding proteins from Haemonchus contortus adult worms partially protects goats against infection during prepatency.

To contribute of the knowledge of the immune mechanisms underlying the response to the immunization of goats with thiol-binding proteins fractions (PBS-TSBP) from Haemonchus contortus (H. contortus) adult worms, this study analyzed the degree of protection and the immune responses developed against the parasite after vaccination with this antigenic complex during the time-elapsing between challenge with L3 of the parasite and the development of adult worms, evidenced by the appearance of first faecal eggs (prepatent period or prepatency). Goat kids immunized with PBS-TBSP generated an immune response during the prepatency which translates into a reduction in the number of worms, as well as a lower reduction on packed cell volume and plasma protein levels in relation to the non-vaccinated animals. As previously described in other studies carried out after the prepatent period, this protection was associated with a systemic humoral response. At the local level, a specific humoral response was also observed, together with an immune-inflammatory infiltrate in the gastric mucosa of MCH-II + cells and CD4+ lymphocytes, whose number was associated with a reduction in the number of worms and an increase in plasma proteins. A high peripheral eosinophilia was detected, but no corresponding increased infiltration of the gastric mucosa by eosinophils or globular leukocytes was observed. In agreement with previous data on the immunolocalization of the antigens used here, the results obtained contribute to the idea that these may be excretion/secretion (E/S) products necessary for parasite survival, whose inactivation during the larval and/or pre-adult stages may have contributed to immunoprotection.

Selective non-antibiotic treatment in sigmoid diverticulitis: is it time to change the traditional approach?

BACKGROUND: A growing body of knowledge is calling into question the use of antibiotics in acute diverticulitis (AD). Moreover, recent studies provide evidence regarding the security of treating patients with AD as outpatients. The aim of this study was to evaluate a restrictive antibiotic outpatient protocol for the treatment of mild-to-moderate episodes of AD. METHODS: All patients with symptoms of AD presenting to our emergency department were assigned a modified Neff stage. Patients with mild AD received outpatient treatment without antibiotics. Patients with mild AD and comorbidities were admitted to receive the same treatment. Patients with moderate AD were admitted for 48 h and were then managed as outpatients until they had completed 10 days of antibiotic treatment. RESULTS: Between April 2013 and November 2014, we attended 110 patients with a diagnosis of AD, 77 of whom we included in the study: 45 patients with mild AD and 32 with moderate AD. Of the patients with mild AD, 88.8 % successfully completed the non-antibiotic, non-admission treatment regime and 95.5 % benefited from a non-antibiotic regime, whether as outpatients or inpatients. A total of 88 % of patients with mild AD and 87.5 % of patients with moderate AD who met the inclusion criteria completed treatment as outpatients without incident. No major complications (abscess, emergency surgery) or deaths were recorded. CONCLUSIONS: Outpatient treatment without antibiotics for patients with mild AD is safe and effective. Patients with moderate AD can be safely treated with antibiotics in a mixed regime as inpatients and outpatients.

Does stapled anopexy for bleeding haemorrhoids cure associated anaemia?

AIM: The aim of this study was to evaluate the effectiveness of stapled anopexy (SA) in patients with chronic bleeding haemorrhoids and secondary anaemia. METHOD: Our department performed 340 SA procedure per patient for haemorrhoids between January 1999 and December 2011. Fifty (14.7%) of these patients (25 male patients and 25 female patients) had anaemia (haemoglobin concentration < 13 g/dl in male patients and < 12 g/dl in female patients) secondary to chronic haemorrhoidal bleeding. Patients with colorectal bleeding and anaemia not caused by haemorrhoids were excluded. The mean (SD) age was 56.4 (13.9) years and the mean (SD) haemoglobin concentration was 9.2 (1.6) g/dl for male patients and 10.4 (1.2) g/dl for female patients. Five (10%) patients with anaemia had Grade II, 22 (44%) had Grade III and 23 (46%) had Grade IV haemorrhoids. The median (range) duration of postoperative follow-up was six (1-12) years. RESULTS: None of the patients required early postoperative admission or experienced early or late complications related to SA. The procedure was successful (normal haemoglobin concentration and no bleeding at 6 months postsurgery) in 45 (90%) patients. Of the five (10%) patients in whom SA was ineffective, one had Grade II, three had Grade III and one had Grade IV haemorrhoids. All these patients underwent Milligan-Morgan haemorrhoidectomy 3 months after SA. CONCLUSION: SA is an effective treatment for patients with bleeding haemorrhoids and subsequent anaemia. In our experience, the success rate was satisfactory and there were no serious complications.

Prevalence of metabolic acidosis in ileal diversions more than one year after radical cystectomy and associated secondary metabolic effects.

INTRODUCTION AND OBJECTIVE: Metabolic acidosis (MA) is a well-known complication in patients with ileal urinary diversions. It is common in the early postoperative stages and decreases over time. Our objective is to investigate the prevalence of MA after more than one year of follow-up, identify the associated risk factors, and analyze its secondary metabolic consequences. MATERIALS AND METHODS: We conducted an observational study between January 2018 and September 2022 following the STROBE guidelines. MA was defined as a serum bicarbonate level ​​<22mEq/L. Finally, we analyzed 133 patients with a mean follow-up of 55.24 ± 42.36 months. RESULTS: MA was observed in 16 (12%) patients. Patients with and without MA were comparable in age, sex, and follow-up time. The group with MA presented a higher rate of anemia (68,75% vs 19,65%, p < 0.001) and renal failure (100% vs 45,29%, p < 0.001), statistically significant higher levels of serum creatinine, chloride, potassium, parathyroid hormone, and phosphorus but lower serum values ​​of hemoglobin, renal glomerular filtration rate, total cholesterol, vitamin D, calcium, and albumin (all p < 0.05). Renal glomerular filtration rate was the only independent risk factor related to the development of MA (OR 0.914; 95% CI 0.878-0.95; p < 0.0001), proving a close correlation with venous bicarbonate values ​​(r = 0.387, p < 0.001). CONCLUSIONS: MA is a little prevalent disorder in ileal urinary diversions more than one year after radical cystectomy is performed but it has secondary consequences on hematologic, renal, protein, lipid, and bone metabolism. We recommend to a close follow-up in patients with renal failure for early diagnosis and treatment.

Day case stapled anopexy for the treatment of haemorrhoids and rectal mucosal prolapse.

AIM: Stapled anopexy (SA) gives better early postoperative results than classical haemorrhoidectomy. The aim of this study is to demonstrate that SA is a safe and effective procedure for the treatment of haemorrhoids and rectal mucose prolapse in a day-case surgery programme. METHOD: From January 2000 to December 2008, 297 SA procedures were performed; 230 (77.4%) were performed in the Day Surgery Unit (DSU). Third- and fourth-degree haemorrhoids, second-degree haemorrhoids with no response to conservative treatment and several cases of rectal prolapse were included. The mean age of the patients in the series was 48.1 years (range 21-85). Preoperative preparation included phosphate enemas and antibiotic prophylaxis. Patients were operated on mainly under spinal anaesthesia. Day-case rate, postoperative pain (measured by a visual analogic scale, 1-10), admissions, re-admissions, early postoperative situation and recurrence were evaluated in the study. RESULTS: The overall DSU rate was 78%, with a progressive increase from 46% to 99% in 2008. One hundred and eighty-five patients (80%) had pain scores under 2; no patient had a pain score over 7. Eighteen (8%) patients required admission on the day of surgery. Late admission was needed for 3 (3%) patients. Thirty-three patients reported their situation as excellent, 174 as good, 20 as acceptable and three as bad when they answered a phone questionnaire 24 h after surgery. Overall, 20 (9%) patients had recurrence of symptoms. CONCLUSION: SA is a safe and effective procedure for prolapsing haemorrhoids in the day case setting. The recurrence rate is higher than that observed in classical haemorrhoidectomy. Most patients can be managed as day-cases.

Presence of C. albidus, C. laurentii and C. uniguttulatus in crop and droppings of pigeon lofts (Columba livia).

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.

Ligation of intersphincteric fistula tract (LIFT) for the treatment of anal fistula: a prospective observational study

Background: The ligation of intersphincteric fistula fract (LIFT) technique avoids postoperative anal continence disturbances and preserves quality of life. Methods: A total of 70 patients with anal fistula (AF) were treated in the Day Surgery Unit. The LIFT technique was the primary treatment in 63 patients. The other had previously undergone placement of a loose seton (two-step approach). The mean follow-up was 66.8 months. Statistical analysis was performed using contingency tables, the chi-square test, and the Student T-test. Results: The use of LIFT was successful in 40 patients (57.1%). However, 6 patients (8.6%) presented persistence of postoperative intersphincteric fistula, being successfully treated by fistulotomy. There were no differences in this technique's success rate between high and low AF (p = 0.45). The success rate of one-step LIFT, however, was significantly higher (p = 0.03). No disturbances of continence were observed. Conclusions: The LIFT technique has a role in the treatment of AF, is suitable for ambulatory surgery, and has a low complications rate. A two-step approach is not always needed. (AU)

Protective immune responses during prepatency in goat kids experimentally infected with Eimeria ninakohlyakimovae.

During the first schizogony, the goat coccidia Eimeria ninakohlyakimovae develops macroschizonts in lacteal duct endothelial cells, whose rupture leads to severe ileal damage and clinical signs during the prepatent period. The immune response elicited against early stages of the parasite development still requires to be investigated. In the present study we have evaluated immune reactions in goat kids primary- and challenged-infected with Eimeria ninakohlyakimovae, and sacrificed during prepatency (7days after challenge). The oocyst output during the primary infection, body weight and clinical condition of all the animals were examined and, at the end of the experiment, all the goat kids were euthanized and subjected to necropsy. Samples were taken from different sections of the ileum, colon and mesenteric lymph nodes (MLN) of primary- and challenged E. ninakohlyakimovae-infected animals. Intestinal leukocyte subpopulations were characterized in E. ninakohlyakimovae-infected mucosa and counts of lymphocytes, eosinophils, polymorphonuclear neutrophils (PMN), globular leukocytes and mast cells were recorded. Additionally, gene expression of caprine IL-2, IL-4, IL-10 and INFγ of ileal, colonic and MLN tissues were performed, as well as the immunohistochemical characterization of immune cells. The E. ninakohlyakimovae primary infection resulted in moderate to severe enteritis with different degrees of diarrhoea and was accompanied by high OPG counts and an increase of most immune cells analyzed when compared to uninfected control animals. Furthermore, eosinophil-, lymphocyte-, globular leukocyte- and mast cell-counts were significantly higher in the challenge group compared to the primary infected animals, whilst the opposite was true for PMN counts. The challenge infection was also associated with moderate increased levels of local mucosal IgA. Interestingly, the number of immature schizonts found at the ileal mucosa was statistically higher in the challenge infected group compared to the challenged control animals. Furthermore, in the challenged E. ninakohlyakimovae-infected animals a significantly higher number of mucosal CD4+ and CD8+ lymphocytes were observed, indicating that these T cell subpopulations might be involved in protective host immune response elicited against early stages of parasite development. The immune response was however very complex, as antigen presenting cells and other effector cell populations of the innate immune system, as well as certain cytokines, were involved. In summary, the results of this study contribute to the better understanding of local cellular and humoral immune responses against caprine E. ninakohlyakimovae, particularly during the prepatency.

Reverse osmosis membrane composition, structure and performance modification by bisulphite, iron(III), bromide and chlorite exposure.

Reverse osmosis (RO) membrane exposure to bisulphite, chlorite, bromide and iron(III) was assessed in terms of membrane composition, structure and performance. Membrane composition was determined by Rutherford backscattering spectrometry (RBS) and membrane performance was assessed by water and chloride permeation, using a modified version of the solution-diffusion model. Iron(III) dosage in presence of bisulphite led to an autooxidation of the latter, probably generating free radicals which damaged the membrane. It comprised a significant raise in chloride passage (chloride permeation coefficient increased 5.3-5.1 fold compared to the virgin membrane under the conditions studied) rapidly. No major differences in terms of water permeability and membrane composition were observed. Nevertheless, an increase in the size of the network pores, and a raise in the fraction of aggregate pores of the polyamide (PA) layer were identified, but no amide bond cleavage was observed. These structural changes were therefore, in accordance with the transport properties observed.

Influence of immunoprotection on genetic variability of cysteine proteinases from Haemonchus contortus adult worms.

The limitations associated with the use of anthelmintic drugs in the control of gastrotintestinal nematodosis, such as the emergence of anthelmintic resistance, have stimulated the study of the immunological control of many parasites. In the case of Haemonchus contortus, several vaccination trials using native and recombinant antigens have been conducted. A group of antigens with demonstrated immunoprotective value are cathepsin B - like proteolytic enzymes of the cysteine proteinase type. These enzymes, which have been observed in both excretory-secretory products and somatic extracts of H. contortus, may vary among different geographic isolates and on strains isolated from different hosts, or even from the same host, as has been demonstrated in some comparative studies of genetic variability. In the present study, we evaluated the genetic variability of the worms that fully developed their endogenous cycle in immunised sheep and goat in order to identify the alleles of most immunoprotective value. To address these objectives, groups of sheep and goats were immunised with PBS soluble fractions enriched for cysteine proteinases from adult worms of H. contortus from either a strain of H. contortus isolated from goats of Gran Canaria Island (SP) or a strain isolated from sheep of North America (NA). The results confirmed the immunoprophylactic value of this type of enzyme against haemonchosis in both sheep and goats in association with increased levels of specific IgG. The genetic analysis demonstrated that the immunisation had a genetic selection on proteinase-encoding genes. In all the immunised animals, allelic frequencies were statistically different from those observed in non-immunised control animals in the four analysed genes. The reduction in the allelic frequencies suggests that parasites expressing these proteases are selectively targeted by the vaccine, and hence they should be considered in any subunit vaccine approach to control haemonchosis in small ruminants.

Direct ultrafiltration performance and membrane integrity monitoring by microbiological analysis.

The feasibility of substituting a conventional pre-treatment, consisting of dioxi-chlorination, coagulation/flocculation, settling and sand filtration, of a drinking water treatment plant (DWTP) by direct ultrafiltration (UF) has been assessed from a microbiological standpoint. Bacterial indicators, viral indicators and human viruses have been monitored in raw river, ultrafiltered and conventionally pre-treated water samples during two years. Direct UF has proven to remove bacterial indicators quite efficiently and to a greater extent than the conventional process does. Nevertheless, the removal of small viruses such as some small bacteriophages and human viruses (e.g. enteroviruses and noroviruses) is lower than the current conventional pre-treatment. Membrane integrity has been assessed during two years by means of tailored tests based on bacteriophages with different properties (MS-2, GA and PDR-1) and bacterial spores (Bacillus spores). Membrane integrity has not been compromised despite the challenging conditions faced by directly treating raw river water. Bacteriophage PDR-1 appears as a suitable microbe to test membrane integrity, as its size is slightly larger than the considered membrane pore size. However, its implementation at full scale plant is still challenging due to difficulties in obtaining enough phages for its seeding.

Study on immune response of goats vaccinated with a live strain of Mycobacterium paratuberculosis.

A study of the immune response of goats vaccinated with a live strain (316-F) of Mycobacterium paratuberculosis has been performed, taking into consideration both cellular and serological responses using a lymphocyte transformation assay (LTA) and a counterimmunoelectrophoresis test (CIET). The analyses made at three monthly intervals reveal an in vitro proliferative response of blood lymphocytes to specific antigen and lymphocytes T mitogen over a year of post-vaccination, but no humoral response was observed when CIET was used.

IgG concentration in mammary secretions of goats throughout lactation in healthy and coagulase-negative staphylococci infected udders.

Radial immunodiffusion was used to determine immunoglobulin (Ig)-G concentrations in 16 mammary secretions from uninfected udder halves and in 10-14 secretions from halves subclinically infected with coagulase-negative staphylococci in goats throughout lactation. IgG concentrations in samples from uninfected halves decreased rapidly during the first week after parturition (P < 0.001 ), henceforth falling slowly up to 30 d post-partum (P < 0.001). From 30 d post-partum to 180 d of lactation, IgG concentration showed a continuous decline to the end of the experiment but these changes were not statistically significant until 150 d after kidding (P < 0.01). IgG concentrations in lacteal secretions of halves harbouring coagulase-negative staphylococci showed a similar pattern, from the first month of lactation, to that observed in healthy udder halves, but concentrations were always lower. These differences were significant from 2 months after parturition (P < 0.001). IgG concentrations were compared with somatic cell counts obtained using an automatic counter (Fossomatic).

The prevalence of Dirofilaria immitis in Gran Canaria, Canary Islands, Spain (1994-1996).

Blood samples from 2034 dogs were tested to detect Dirofilaria immitis antigen during three consecutive years (from 1994 to 1996) in Gran Canaria Island, Canary Islands, Spain. The prevalence of heartworm infection was 67.02% in 1994, 58.92% in 1995 and 52.18% in 1996, with a mean prevalence of 58.89%. Heartworm infection was more common in males (56.19%) than in females (43.81 %), in dogs aged between 3 and 6 years old. The distribution of the disease in the different climatic zones was studied. Chronological changes in the dog's prevalence for heartworm infection in the three consecutive years and the role of the epidemiological factors in the changes of the positive rates were evaluated.

Use of an enzyme-linked immunosorbent assay for serodiagnosis of clinical paratuberculosis in goats. Study by western blotting of false-positive reactions.

An enzyme-linked immunosorbent assay (ELISA) was performed for diagnosis of paratuberculosis in goats, using as antigen a protoplasmic extract (PPA-3). The test was developed on the basis of the results obtained with two serum reference pools, positive and negative respectively. To avoid day-to-day variations, dilutions of the positive serum pool were included in each plate to obtain an arbitrary system, transforming absorbance into immunoglobulin (Ig) G anti-Mycobacterium paratuberculosis units. The ELISA was used on sera of two reference groups of animals. One group consisted of 35 goats suspected of being infected with paratuberculosis, which was confirmed by histological findings and isolation of M. paratuberculosis. The negative group consisted of 61 healthy goats from a farm free of paratuberculosis. The test showed a sensitivity of 100% and a specificity of 91.8%. Absorption of sera with a Mycobacterium phlei suspension did not modify either the sensitivity or the specificity of the test. Sera from the negative group were analysed by Western blotting, and four of them recognized two fractions with a molecular weight of 17.3 and 28.1 kDa.

Immunohistochemical detection of Dirofilaria immitis using a rabbit polyclonal anti-serum against adult somatic antigens and excretory-secretory products.

An immunohistochemical study on tissue sections of kidney, lung, liver and spleen from six dogs naturally infected by Dirofilaria immitis was carried out, using a rabbit polyclonal anti-serum against somatic antigen (SA) of adult worms, excretory-secretory products (ES) of adult worms and fraction C3 of complement (Dog C3). Positive immunoreactions were observed on circulating microfilariae when anti-SA and anti-ES were used, but no differences were noted between the two anti-sera. No antigen deposits were observed when the three anti-sera (anti-SA, anti-ES or anti-C3) were employed as primary antibody. In two cases of human filariosis (by Loa loa and Mansonella streptocerca) no positive immunoreactions were observed.

Pergolide treatment for Cushing's syndrome in a horse.

A 16-year-old, male, Hanoverian horse had a three-month history of weight loss, hirsutism and polyuria/polydypsia. Examinations revealed neutrophilia, lymphopenia, hyper glycaemia and abnormalities in hepatic function. A tentative diagnosis of hyperadrenocorticism was made. The results of thyroid-releasing hormone and combined dexamethasone suppression and ACTH stimulation tests suggested the presence of a pituitary adenoma. The horse was treated with pergolide and beneficial clinical and biochemical responses were observed within one to six months.

[Silent cerebral infarct in patients with sickle cell anemia]. / Infarto cerebral silente en pacientes con anemia falciforme.

INTRODUCTION: Ictus is a complication in patients with Sickle Cell Anemia (SCA), in these is of importance the precocious diagnosis of Silent Cerebral Infarcts (SCI). OBJECTIVE: To determine the incidence of ICS in patients with SCA without neurological symptoms but with images in Cerebral Magnetic Resonance (RMC). PATIENTS: A total of 18 patients (13 males, 5 females) with ages between 5 and 24 years (11.5 4,9), without history of neurological alterations, taken care at the Instituto Hematol gico de Occidente Banco de Sangre, Estado Zulia Venezuela. Methods. A clinical history was made to each patient in addition to detailed physical and neurological examinations that included the state of mind, conscience, language, sensitivity, cranial pairs, muscular force, reflexes, cerebella tests, neck and march. Later, RMC studies were carried out. RESULTS: It was found that 2/18 (11,1%) patients without neurological manifestations showed alterations in the RMC and they were diagnosed as ICS. The findings by images showed asymmetry of lateral ventricles and one of them showed gliosis as well. CONCLUSION: 11,1% of the all the studied cases (2/18) showed SCI, for what is suggested to carry out neurological evaluation and images, once a year, and to offer opportune therapies, for their impact in the function neurocognitive

[Cerebrovascular disease in sickle cell anemia patients]. / Enfermedades cerebrovasculares en pacientes con anemia falciforme.

The cerebrovascular disease is a common complication in Sickle Cell Anemia (SCA). The objective of the present work was to determine the incidence of cerebrovascular disease in patients with SCA, through physical and radiological examination. Twenty one patients between 5 and 18 years were studied, homozygous SS, with or without history of neurologic manifestations, at the "Instituto Hematológico de Occidente, Maracaibo-Venezuela". A clinical history, neurological clinical examination and Cerebral Magnetic Resonance (CMR) were carried out on each patient. CMR showed findings of cerebrovascular disease (CVD) of the ischemic type, in 5 of 21 (23.8%) patients, 3 of them presented neurological alterations (2 with spastic hemiparesis and one with spastic tetraparesis); the other 2 presented silent CVD). The most affected artery was the cerebral media. We suggest to carry out a neurological evaluation (clinic and images) in patients with SCA, once a year, in order to detect early abnormalities of the central nervous system and to offer timely and adequate therapies, avoiding important sequels for the patient normal life development.

Immunoprotective effect of cysteine proteinase fractions from two Haemonchus contortus strains adapted to sheep and goats.

A preliminary analysis of the significance of genetic diversity in cysteine proteinase genes has been performed simultaneously in sheep and goats, with regard to the immunological control using these enzymes against haemonchosis. For this purpose, we have studied the cross-immunoprotective effect of cysteine protease-enriched protein fractions (CPFs) in adult worms of two Haemonchus contortus strains from North America and Spain that are adapted to sheep and goats, respectively. Previous genetic analysis of cysteine proteinase genes in both strains has shown that some of loci are polymorphic and these differences are translated into changes in the amino acid sequences. However, our results show that CPFs from H. contortus adult worms have a protective effect against the parasite in both sheep and goats. These results are similar regardless of whether they were obtained from sheep or goat-adapted H. contortus strains, which could be very important in case H. contortus CPFs were commercially used in different countries, as vaccines to prevent the negative effects of this parasite. Interestingly, this experimental inoculation of both species with a heterologous strain of H. contortus contributes to the idea shown in previous studies about how difficult is the interpretation and the comparison of vaccination where strains not adapted to a specific host are used. Therefore, the challenger of using heterologous strains could provide similar results to those observed in immunised animals. This study suggests the possibility of exploring the mechanisms involved in natural protection against non-adapted strains, in order to develop strategies to control haemonchosis.